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Aldehyde-fuchsin: historical and chemical considerations.

The staining mechanisms of Gomori's aldehyde-fuchsin are not yet fully understood. It seemed therefore timely to review the history of this dye class in context with current dye and aldehyde chemistry. In 1861 Lauth treated basic fuchsin with acetaldehyde. This dye became known as Aldehyde Blue, but consisted of violet and blue dyes. Schiff (1866) studied several aldehyde-fuchsins; these compounds contained two molecules of dye and three molecules of aldehyde. Acetaldehyde-fuchsin prepared according to Schiff's directions showed staining properties similar to those of Gomori's aldehyde-fuchsin. This dye class was soon superseded by new dyes more suitable for textile dyeing, and chemical investigations of aldehyde-fuchsins ceased around the turn of the century. Gomori's aldehyde-fuchsin has been regarded as a Schiff base. However, according to chemical data, low molecular aliphatic aldehydes and aromatic amines tend to form condensation products. Correlations of chemical and histochemical observations suggest such processes during aging of dye solutions. Models of dimers and polymers of aldehyde-fuchsin could be built without steric hindrance. The nature of the bonds formed by various components of aldehyde-fuchsin solutions is not clear. However, cystine in proteins, e.g. in basement membranes, apparently does not play a role in the binding of aldehyde-fuchsin by unoxidized Carnoy- or methacarn-fixed sections.

Aldehydes↗

Pseudo second order kinetics and pseudo isotherms for malachite green onto activated carbon: comparison of linear and non-linear regression methods.

Pseudo second order kinetic expressions of Ho, Sobkowsk and Czerwinski, Blanachard et al. and Ritchie were fitted to the experimental kinetic data of malachite green onto activated carbon by non-linear and linear method. Non-linear method was found to be a better way of obtaining the parameters involved in the second order rate kinetic expressions. Both linear and non-linear regression showed that the Sobkowsk and Czerwinski and Ritchie's pseudo second order model were the same. Non-linear regression analysis showed that both Blanachard et al. and Ho have similar ideas on the pseudo second order model but with different assumptions. The best fit of experimental data in Ho's pseudo second order expression by linear and non-linear regression method showed that Ho pseudo second order model was a better kinetic expression when compared to other pseudo second order kinetic expressions. The amount of dye adsorbed at equilibrium, q(e), was predicted from Ho pseudo second order expression and were fitted to the Langmuir, Freundlich and Redlich Peterson expressions by both linear and non-linear method to obtain the pseudo isotherms. The best fitting pseudo isotherm was found to be the Langmuir and Redlich Peterson isotherm. Redlich Peterson is a special case of Langmuir when the constant g equals unity.

Adsorption↗

Patent blue V encapsulation in liposomes: potential applicability to endolympatic therapy and preoperative chromolymphography.

The water soluble dark blue stain Patent Blue V was incorporated into liposomes to increase its depot properties. Experiments with intraperitoneal and intramuscular application in rats and with intralymphatic injections in rabbits showed a substantial difference between the pharmacokinetics of aqueous and liposomal solutions. Independent of the route of administration, the dye-stain remained much longer in the tissues using liposomes. Most notably, retroperitoneal lymph nodes in rabbits remained dark blue up to 28 days after hindlimb endolymphatic instillation of liposomal patent blue. This vastly superior depot effect may be clinically adaptable for endolymphatic therapy with cytostatic drugs to enhance nodal drug retention and concentration or for preoperative chromolymphography to facilitate nodal visualization at laparotomy.

Animals↗

A methodology for determination of phospholipids.

A simple method for the determination of phospholipids in an aqueous dispersion and in amniotic fluid was developed. The procedure is based on the observation that dispersed phospholipids promoted the solubilization of an insoluble dye--detergent complex. The solubilization of the complex between the negatively charged dye, Coomassie brilliant blue (CBB), and a positively charged detergent, cetyltrimethylammonium bromide (CTAB), produced a blue solution having a visible absorbance maximum above 600 nm. A linear increase in absorbance intensity occurs with an increase in phospholipid concentration. An assay using the CBB-CTAB reagent adsorbed on 3-mm glass beads is used to estimate total dispersed phospholipids between 2 and 25 micrograms/ml. Thereby, a two-phase water-methanol-chloroform system is formed. The products of zwitterionic phospholipids (such as phosphatidylcholine and phosphatidylethanolamine) partition to the organic phase while the dye complex solubilized in anionic phospholipids (such as phosphatidylglycerol and phosphatidylinositol) partitions to the aqueous phase. This procedure results in a convenient, sensitive, and rapid method for the simultaneous determination of the total phospholipid, zwitterionic phospholipid, and anionic phospholipid concentrations. Application of the new assay for determination of phospholipids in amniotic fluid is described.

Amniotic Fluid↗

Preliminary study of sentinel node identification with 99mTc colloid and blue dye in patients with endometrial cancer.

AIMS AND BACKGROUND: Intraoperative lymphatic mapping and sentinel node (SLN) biopsy have generated a tremendous amount of interest and are already established as part of the standard practice in the surgical management of breast cancer and melanoma. To reduce extensive radical procedures and decrease the morbidity in the treatment of gynecologic malignancies, much effort is being made to use less aggressive interventions. The purpose of our study was to determine the feasibility of SLN mapping in a group of patients with endometrial cancer at early stages. METHOD AND STUDY DESIGN: Between September 2000 and May 2001 11 patients with endometrial cancer FIGO stage Ib (n = 10) and IIa (n = 1) underwent laparoscopic SNL detection during laparoscopy-assisted vaginal hysterectomy with bilateral salpingo-oophorectomy and bilateral systematic pelvic lymphadenectomy. Radioactive isotope injection was performed 24 hours before surgery and blue dye injection was performed just before surgery in the cervix at 3,6, 9 and 12 hours. A 350 mm laparoscopic gamma scintyprobe MR 100 type 11, 99mTc settled (Pol Hi Tech), was used intraoperatively for SLN detection. RESULTS: Seventeen (17) SLNs were detected with lymphoscintigraphy (six bilateral and five unilateral). At laparoscopic surgery we found the same locations belonging at internal iliac lymph nodes (the so-called Lebeuf-Godard area, lateral to the inferior vesical artery, ventral to the origin of the uterine artery and medial or caudal to the external iliac vein). Fourteen (14) SLNs were negative on histological analysis and three were positive for micrometastases (mean SLN sections = 60). All other pelvic lymph nodes were negative at histological analysis. The same SLN locations detected with the gamma scintyprobe were observed at laparoscopy after patent blue dye injection. CONCLUSIONS: Our preliminary data suggest that combined 99mTc-labeled colloid and vital blue-dye techniques are feasible for SLA detection in endometrial cancer; they represent a very promising tool to transform the management of early-stage endometrial cancer. The clinical validity of this combined technique should be evaluated prospectively.

Adult↗

Microleakage of root restorations.

This study evaluated the microleakage of various restorative materials placed in root surfaces. A minimum of 20 freshly extracted single-rooted teeth were used for each combination of restorative materials. Four preparations were made on the root surface and each restored with a different material. After thermocycling in dye, the root was cut transversely in several sections through the restoration, and microscopically examined to record the microleakage at the interface between restorative materials and tooth. Results indicated that fewer composite resin specimens allowed microleakage into dentin as compared with either amalgam or glass ionomer materials.

Composite Resins↗

[The lens of the eye and its relation to the lymphatic system].

On the basis of well-tried methods of investigation with vital dyes (patent blue violet and Japanese ink) and in combination with the drug (Conjunctisan A), the natural pathways of resorption through the lymphatics of the eye in the mouse, were displayed and this permitted the transport mechanisms to be followed. It was possible gradually to understand the morphology of the lymphatic drainage and, in this way evidence was produced regarding the relationships of the lymphatic system and the regional lymph nodes.

Animals↗

Standardization of the Coomassie Blue method for cerebrospinal fluid proteins.

Coomassie Brilliant Blue G-250 can be used to quantitatively determine proteins in cerebrospinal fluid. When the dye combines with protein, the absorption maximum of the dye shifts. The dye-protein color forms almost instantaneously and is stable for at least 1 h. The procedure is also insensitive to changes in temperature in the range of 20--30 degrees C. The absorptivities of the dye complexes with human albumin or globulin differ, thus a pure albumin or pure globulin standard is unsuitable; a standard containing 70% albumin and 30% globulins is the most appropriate for this application. A bichromatic approach to standardization increases the range of linearity of a calibration curve. The method gives values that are about 9% higher than a sodium sulfate-sulfosalicylic turbidimetric procedure for cerbrospinal fluid proteins.

Cerebrospinal Fluid Proteins↗

1% lymphazurin vs 10% fluorescein for sentinel node mapping in colorectal tumors.

HYPOTHESIS: Ten percent fluorescein may be successfully used as an alternative to 1% Lymphazurin (1% isosulfan blue; US Surgical Corp, North Haven, Conn) in sentinel lymph node (SLN) mapping for the accurate staging of colorectal tumors. DESIGN: Review of prospectively gathered data. SETTING: University-affiliated regional medical center. PATIENTS: Sentinel lymph node mapping was performed in 120 consecutive patients with colorectal malignancies. INTERVENTIONS: The first 1 to 4 blue nodes detected within 5 minutes were designated as Lymphazurin-detected SLNs. The first 1 to 4 fluorescent nodes seen under the Wood light were designated as fluorescein-detected SLNs. Multilevel serial sections for hematoxylin-eosin and immunohistochemistry studies for cytokeratin were performed on all SLNs. MAIN OUTCOME MEASURES: Successful mapping, accuracy, skip metastasis, adverse reactions, occult micrometastases detection, and cost. RESULTS: Mapping was successful using Lymphazurin in 99% of the patients vs 97% of the patients using fluorescein (P =.89). The accuracy of predicting nodal metastases with each tracer was 95.8% vs 93.1%, respectively (P =.82). The skip metastases rate was 4.2% for Lymphazurin vs 6.9% for fluorescein (P =.37). The 5 patients in whom nodal disease was only identified as occult micrometastasis in the SLNs had a total of 5 SLNs, all of which were identified by both tracers. No adverse reactions occurred. The cost for Lymphazurin was $99.00, while the cost for fluorescein was $2.10. CONCLUSIONS: With the exception of cost, there were no statistically significant differences between the 2 dyes. While easy availability and lower cost remain distinct advantages of fluorescein, Lymphazurin remains the gold standard. In patients with known hypersensitivity to Lymphazurin and when availability and cost are an issue, fluorescein may be used effectively for SLN mapping in colorectal tumors.

Adult↗

Lymphatic mapping and sentinel lymphadenectomy for primary and metastatic pulmonary malignant neoplasms.

BACKGROUND: Mediastinal lymph node sampling understages a significant number of lung cancers, even when nodes are evaluated by immunohistochemical techniques. Intraoperative lymphatic mapping and sentinel lymphadenectomy allows focused pathologic evaluation of a few lymph nodes that accurately stage the entire basin. HYPOTHESIS: Lymphatic mapping and sentinel lymphadenectomy is a practical and accurate method of staging lymph nodes that drain primary and metastatic neoplasms of the lung. DESIGN AND SETTING: Retrospective review at a tertiary referral center. PATIENTS: Sixty-seven patients undergoing resection of lung tumors. MAIN OUTCOME MEASURES: Sentinel lymph node (SN) identification rate, number of SNs, nodal pathologic features, and survival. RESULTS: Twenty-eight patients had primary lung cancer and 39 had pulmonary metastases from melanoma (33 cases), squamous cell carcinoma (2 cases), colon cancer (2 cases), or other cancers (2 cases). Lymphatic mapping and sentinel lymphadenectomy was successful in all patients. The median number of lymph nodes identified by dye alone was 2 (range, 1-7); the median number identified by dye plus radiocolloid was 4 (range, 1-9). Most SNs (69%) were N1; 31% were N2. Lower lobe lesions drained to upper mediastinal nodes in 3 (13%) of 24 cases. Lymph node metastases were found in 11 patients with lung cancer (39%) and 8 patients with pulmonary metastases (21%). Ten (91%) of the 11 patients with lung cancer had SN involvement. In the 33 patients with metastatic melanoma, SN involvement significantly reduced the rate of 2-year survival (0% vs 48%). CONCLUSIONS: Lymphatic mapping and sentinel lymphadenectomy of intrapulmonary malignancies is technically challenging but feasible. Blue dye is most useful for in vivo identification of SNs; ex vivo radioactivity can confirm that excised nodes are SNs. Lymphatic mapping and sentinel lymphadenectomy can provide important prognostic information for patients with melanoma and lung metastases, and it may improve the staging of primary lung cancer.

Adult↗

Electrophoretic pattern of rodent seminal vesicle proteins as revealed by silver staining.

Electrophoresis of seminal vesicle secretions (SVS) from several rodents showed a very simple pattern composed of 3-5 main protein bands when an anionic dye (Coomassie brilliant blue) was used. However, use of a silver staining method showed a more complex protein spectrum, and several minor components of 12-90 kD, were clearly revealed. Western blotting using antibodies to SVS demonstrated that these minor protein components were not serum contaminants. Rat, mouse and hamster SVS shared antigenic determinants which were not related to rabbit SVS.

Animals↗

Biotransformation of malachite green by the fungus Cunninghamella elegans.

The filamentous fungus Cunninghamella elegans ATCC 36112 metabolized the triphenylmethane dye malachite green with a first-order rate constant of 0.029 micromol x h(-1) (mg of cells)(-1). Malachite green was enzymatically reduced to leucomalachite green and also converted to N-demethylated and N-oxidized metabolites, including primary and secondary arylamines. Inhibition studies suggested that the cytochrome P450 system mediated both the reduction and the N-demethylation reactions.

Biodegradation, Environmental↗

Marginal microleakage in bonded amalgam restorations: a combined in vivo and in vitro study.

PURPOSE: This study evaluated the marginal microleakage in bonded amalgam restorations in teeth extracted a month after restoration placement. MATERIALS AND METHODS: Nine teeth in a patient scheduled for complete extraction were selected. In each tooth, a buccal prepared cavity was treated with Scotchbond Plus and filled with high copper amalgam. In four teeth, a lingual unlined restoration was also prepared as control. A month later, all the teeth were extracted; seven teeth were immersed for 48 hours in a 0.05% basic fucsine dye solution and then sectioned. Sections were observed with a stereomicroscope at 10X. Results were statistically evaluated using a nonparametric Mann-Whitney test for independent data. The remaining two teeth were sectioned and observed using a scanning electron microscope (SEM). RESULTS: Stereomicroscopic analysis revealed that 80% of bonded specimens did not exhibit dye infiltration at the enamel margin; the same percentage of bonded specimens showed partial infiltration at the cervical margin. In the unlined control specimens, there was a complete dye infiltration at both margins. SEM analysis revealed a good adaptation between resin and tooth and between resin and amalgam. CONCLUSION: Bonded amalgam restorations are effective in reducing marginal microleakage, particularly at the enamel margin.

Aged↗

An evaluation of elastic tissue staining.

The Special Procedures Laboratory of the Department of Pathology at M.D. Anderson Hospital was concerned with the nationwide shortage of hematoxylin. The amount of this dye required for various staining solutions was calculated to determine thrifty usage. Some solutions required more hematoxylin than was available and prompted the serious investigation of existing techniques and new methods for routine laboratory use. Evaluation of various techniques resulted in a significantly successful modification of Gomori's aldehyde fuchsin technique, which was subsequently adopted as a routine procedure for elastic tissue staining in the Special Procedures Laboratory.

Aldehydes↗

Reliability of identification of 655 sentinel lymph nodes in 263 consecutive patients with malignant melanoma.

BACKGROUND: The purpose of this study of sentinel lymph node biopsies (SLN) was threefold: to compare the reliability of lymphazurin blue dye to radioactive technetium 99m sulfur colloid (TC); to evaluate the reliability of frozen section examinations of sentinel lymph nodes; and to determine how much SLN dissections prolonged operative time. STUDY DESIGN: We evaluated the records of 263 consecutive patients with intermediate and high-risk melanomas (1.0 mm or thicker, or Clark Level IV or greater), who were treated by a single surgeon at the Yale Melanoma Unit between October 1, 1997, and September 30, 2001, and followed for more than 18 months. RESULTS: A total of 655 SLN were identified and removed from these 263 consecutive patients. Radioactive colloid was found to be more reliable (100%) in identifying the SLN than lymphazurin blue dye (51%) in the nodes of the patients. Twenty-eight patients (11%) had positive sentinel lymph nodes, and 2 patients (7%) had false-negative frozen sections. Three patients (11%) had false-negative frozen sections; tumor was found subsequently on permanent sections only after special immunohistochemical stains were used. The location or removal of SLN did not prolong the operative procedure unreasonably, requiring an average of 7 to 20 minutes for removal of SLN, and 33 minutes for frozen section reports, during which time the primary tumor resection and wound coverage were performed. CONCLUSIONS: SLN were found in all 263 patients. All SLN were identified reliably with radioactive colloid. Because blue dye was found in only half of the radioactive nodes, it is not appropriate to use this as the only marker for locating the SLN. This large series of patients attests to the reliability of frozen sections in identifying SLN harboring metastases, with 82% of the patients with nodal metastases identified in this fashion.

Follow-Up Studies↗

Sentinel node biopsy for the detection of head and neck melanoma: a review.

Worldwide incidence of malignant melanoma is on the rise. Early detection of this malignancy is key to survival, and in the case of more advanced malignancy, early and effective detection of micrometastatic disease is crucial for staging and therapy. Because melanoma spreads primarily via lymphatic drainage patterns, effective methods for tracing these pathways are of paramount importance. The authors summarize the efficacy of blue dye, gamma probe, and lymphoscintigraphy detection methods, both individually and combined; the "missed disease" (or false-negative) rate; and the clinical discordance between expected and actual location of metastatic disease in head and neck melanoma. A clinical meta-analysis of current studies in head and neck melanoma was used to evaluate clinical data. A success rate of 95% to 100% for detection of sentinel lymph nodes can be achieved when blue dye, gamma probe, and lymphoscintigraphy techniques are combined. This is associated with a false-negative rate of 7.7% to 10.4%. With respect to intermediate-depth melanomas of the head and neck, a significant discordance exists between expected and actual lymphatic drainage patterns. This problem is best addressed using a combination of lymphoscintigraphy, blue dye, and gamma probe localization, which yields a success rate of 95% to 100% for detection of sentinel lymph nodes and a low false-negative rate of 7.7% to 10.4%. In the instance of a failed study, one in which sentinel nodes are not detected by the aforementioned methods, elective node dissection is the treatment modality of choice.

False Negative Reactions↗

[Sentinel node navigation surgery in uterine cancer].

The Clinical test of sentinel nodes (SNs) in uterine cancer began recently, especially endometrial cancer. In relation to cervical cancer, the detection rate of SN ranged anywhere between 15% to 100% and likewise the sensitivity rate varied from 66% to 100%. Due to these inconclusive results, the data cannot be reliably used for clinical study purposes. Meanwhile, it should be noted that endometrial cancer research has just begun. The tracer used for detection can be roughly classified into an isotope method and a dye method. The isotope method shows a better detection rate, but it can supplemented by using together with the dye method. In Japan, the most commonly reported radiopharmaceutical agent is technetium-99m-labeled phytic acid isotope and 1% isosulfan blue as the dye. In conducting cervical cancer studies, there are many reports of utilizing technetium-99m-labeled phytic acid isotope, injected into 4 different areas of the cervix. Regarding research conducted for endometrial cancer, experiments range from direct injection into the uterine body, injections into the cervix, and hysteroscopic injection. Due to ambiguity in the results, additional clinical testing is required to gather more data.

Female↗

Retroperitoneal and lateral pelvic lymphadenectomy mapped by lymphoscintigraphy and blue dye for rectal adenocarcinoma staging: preliminary results.

BACKGROUND: Total mesorectal excision (TME) is the standard surgical choice for rectal adenocarcinoma. Better prognostic results, achieved with a retroperitoneal and lateral pelvic lymphadenectomy (RLPL), have questioned that TME might not be satisfactory for adequate patient staging, affecting therapeutic definitions. The aims of the ongoing study are to define the accuracy of dye and probe search in the detection of metastatic retroperitoneal and lateral pelvic nodes (RLPN) resected with RLPL, and to evaluate the metastasis frequency in these nodes and its eventual upstaging impact. METHODS: Thirty rectal adenocarcinoma patients were submitted to RLPL, with RLPN mapping using technetium (Tc 99 m) and patent blue, having nodes examined histopathologically and immunohistochemically. RESULTS: Eight hundred and two nodes were analyzed, mean of 26.7 per patient; RLPL was responsible for 41% (330) of the examined nodes, mean of 11 per patient. Metastatic RLPN have occurred in 20% of the patients; the RLPN were metastatic in only 6.7% of the patients; RLPL upstaged 13.3%. For identification of metastatic RLPN with technetium, sensitivity was 33%, specificity 79%, positive predictor value (PPV) 29%, negative predictor value (NPV) 83% and false negative (FN) rate 67%. For patent blue and technetium metastatic RLPN identification, sensitivity was 17%, specificity 92%, PPV 33%, NPV 82% and FN 83%. CONCLUSIONS: Preliminary results have pointed out technetium and blue dye low accuracy to identify metastatic RLPN; no metastatic RLPN were reported in the patients submitted to preoperative chemoradiation and important upstaging with RLPL. Considering no increase in morbi-mortality rates with RLPL, definitive conclusions will be obtained as the study carries on.

Adenocarcinoma↗