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PubMed2001

Effects of flowable composite lining and operator experience on microleakage and internal voids in class II composite restorations.

STATEMENT OF PROBLEM: When inexperienced clinicians perform class II composite restorations, improper placement techniques can lead to problems, including marginal adaptation and void formation. PURPOSE: The aim of this study was to determine the influence of flowable composite linings on marginal microleakage and internal voids in class II composite restorations performed by practitioners with different levels of experience. MATERIAL AND METHODS: Eighty extracted molars were prepared with mesial and distal class II cavity preparations and divided into 4 groups. Each group was restored separately with the following materials: Prodigy/Revolution lining (group I), Prodigy (group II), Tetric Ceram/Tetric Flow lining (group III), and Tetric Ceram (group IV). Each group was equally divided and restored by 2 practitioners, one experienced and another untrained in composite restorations. After restoration, all teeth were stored for 24 hours, thermocycled (at 5 degrees C to 60 degrees C) 1500 times, and soaked in 2% basic Fuchsin dye for 24 hours. After soaking, the teeth were sectioned, and gingival marginal microleakage and internal voids (at the gingival wall interface and in the cervical and the occlusal parts) were recorded. Data were analyzed with the Kruskal-Wallis test. RESULTS: There was no significant reduction in microleakage for either practitioner. There were fewer interface voids within pairs with or without flowable composite linings made by the experienced practitioner (P<.05). CONCLUSION: When flowable composite lining was placed at the gingival floor of a class II composite restoration by an experienced practitioner, voids in the restored interface were reduced. Gingival marginal sealing was not improved by the same technique.

Composite Resins↗
PubMed1984

Toxicological studies on malachite green: a triphenylmethane dye.

The oral LD50 for malachite green oxalate was found to be 275 mg/kg in rats while the approximate lethal dose for NMRI mice was 50 mg/kg. No systemic effects were seen after dermal application of 2,000 mg/kg. Repeated administration in the diet for 28 days to rats produced only minor changes in serum urea and aspartate aminotransferase levels. The rats at the highest dose level showed decreased weight gain and appeared clinically to have elevated motor activity. No sex differences were observed in either acute or prolonged experiments. In accord with human experience malachite green was irritating to mucous membranes, but no effects were seen on intact skin nor was it shown to be sensitizing. It was found to be a mutagen in the Salmonella/microsome test after metabolic activation but without clastogenic activity when tested at maximally tolerated levels in mice in the micronucleus test.

Animals↗
PubMed2004

[Lymph nodes mapping for detection of sentinel nodes in patients with papillary thyroid cancer].

We examined the feasibility of sentinel lymph node biopsy for papillary thyroid cancer. In the dye injection method, 1% of isosulfan blue dye was injected around the tumor of 32 patients intra-operatively, and in the radioisotope (RI) colloid injection method, 99mTc-tin colloid was injected in 23 patients 1 day preoperatively. Lymph node mapping for detection of sentinel nodes was performed after thyroidectomy and central and modified lateral neck lymph node dissections. All dissected nodes were examined postoperatively by hematoxylineosin staining to determine whether or not metastasis was present. In the dye injection method, sentinel lymph nodes were identified in 30 (94%) of the 32 patients. Lymph node metastases were found in 14 patients, and some sentinel lymph nodes had papillary cancer metastasis in 13 patients. There was only 1 false-negative case. Sensitivity and accuracy of sentinel lymph node biopsy was 93% (13/14) and 97% (29/30). With the RI method, detection rate, sensitivity and accuracy of sentinel lymph node biopsy was 96% (22/23), 90% (9/10) and 95% (21/22), respectively. Our preliminary study indicated that sentinel lymph node biopsy was feasible in patients with thyroid cancer. It may be helpful in avoiding unnecessary lymph node dissection and improving quality of life in patients with thyroid cancer.

Carcinoma, Papillary↗
PubMed2002

Local pharmaceutical release from a new hydrogel implant.

BACKGROUND: Solid hydrogel polymers can act as reservoirs for controlled drug release. The object of this study was to quantify release kinetics for a single example of a class of uniquely structured hydrogels. The polymer of this study belongs to a class of permanent implants that release pharmaceuticals by diffusion from an entangled cross-linked matrix rather than by absorption of the implant by the body. The cross-linked matrix (CLM) of this study is biocompatible and polymerized in situ, forming a solid that is mechanically bonded to the implant site. It can seal tissue as well as deliver drugs at predetermined rates. We evaluated dye and antibiotic egress and assessed release kinetics and retention of antibiotic activity following elution from the CLM. MATERIALS AND METHODS: The prepolymerized test CLM was prepared in two ratios, 6 and 20% prepolymer, in an activating aqueous solution. Aqueous solutions were prepared from dyes of varying molecular weights. Aliquots of prepolymerized test CLM were allowed to solidify and dye release into the supernatant was quantified by spectrophotometry over a 168-h period. Antibiotic solutions were also employed to form solid CLMs. Tetracycline release over time was characterized by spectrophotometry. Antibiotic dosed solid CLMs were placed on agar plates streaked with Escherichia coli and incubated. Growth inhibition was assessed for each antibiotic. RESULTS: In the test CLM, dye and antibiotic release were found to be inversely related to molecular weight and consistent with a diffusion model. CLMs formed from aqueous solutions containing higher molecular weight dyes and antibiotics released those constituents more slowly than lower molecular weight constituents. This finding, as well as the effect on release rate under varying prepolymer concentration, was consistent with a diffusion release mechanism. Antibiotic released from the tissue sealant was shown to be potent by consistent inhibition of E. coli. CONCLUSIONS: Pharmaceutical release by a representative CLM was found to be controllable by varying the concentration of the pharmaceutical in the activating aqueous solution. The polymerization and release mechanisms did not degrade antibiotic biologic activity. CLMs may be a general class of biocompatible polymers that can locally deliver clinically useful biologics, the release kinetics of which are unaffected by the variability of implant absorption/inflammation mechanisms.

Coloring Agents↗
PubMed1983

A four- to sixfold enhancement in sensitivity for detecting trace proteins in dye or silver stained polyacrylamide gels.

A very simple procedure which enhances sensitivity in visualizing proteins stained by all standard procedures in polyacrylamide gels is described. The process, based on the use of concentrated polyethylene glycol 6000 solutions, produces a conspicuous, reversible, and uniform size reduction of gels. The entity of the reduction depends principally on the polyethylene glycol 6000 concentration and on the acrylamide content of gels. Using an appropriate polyethylene glycol 6000 concentration, it is possible to regulate the final size and then the sensitivity as needed. Depending upon the above conditions it is possible to increase the sensitivity by a factor of 4-6 or more. Reduction of the gel volume by such a factor may eliminate the need for the cumbersome and frequently impossible preconcentration of samples. Besides the Coomassie blue stained gels, silver stained gels may also be treated in a similar manner with a consequent further enhancement in sensitivity. In addition, gels containing radiolabeled materials can be autoradiographed in times proportionally shorter, without significant sacrifice of resolution.

Coloring Agents↗
PubMed2001

Evaluation of feasibility and accuracy of sentinel node biopsy in early breast cancer.

BACKGROUND: Current literature has suggested that sentinel node biopsy may eventually replace axillary dissection as the nodal staging procedure of choice in early breast cancer. The goals of our study were to determine the accuracy of the sentinel node in predicting axillary nodal status and to evaluate the feasibility of incorporating sentinel node biopsy into a general surgical practice. METHODS: Between June 1999 and August 2000, 158 clinically node negative women with a histological diagnosis of T1 or T2 breast cancer were enrolled in the study. Both technetium sulfur colloid radiotracer and isosulfan blue dye were used to guide sentinel node biopsy. Sentinel node biopsy was always followed by a complete axillary dissection. The histopathology of sentinel nodes using serial sectioning and cytokeratin immunohistochemistry was compared with that of the nonsentinel nodes evaluated with routine hematoxylin and eosin stain. RESULTS: The overall sentinel node detection rate was 84% (89 of 106 patients). Sentinel node biopsy was most successful when a combination of radiotracer and dye was used. The staging accuracy of sentinel node biopsy was 98% (87 of 89); the sensitivity of the method was 94% (34 of 36); the false negative rate was 6% (2 of 36); the negative predictive value was 96% (53 of 55); and the rate of metastases to the sentinel node only was 56% (20 of 36). The results varied considerably among surgeons. CONCLUSIONS: The findings in our study support the hypothesis that the sentinel node is an accurate predictor of axillary nodal status in women with early breast cancer. These results suggest that the excellent findings in the literature can be reproduced by a group of general surgeons in a community-based hospital.

Adult↗
PubMed2001

The sentinel node procedure with Patent Blue V dye in the surgical treatment of papillary thyroid carcinoma.

How far to extend the surgical treatment of papillary thyroid carcinoma (PTC) is still an open question. A contribution may come from intra-operative lymphatic mapping because, in other malignancies, the procedure has become an important aid in defining lymph node status. To assess the feasibility of using the sentinel lymph node (SLN) technique with the intratumoral injection of Patent Blue V dye to guide nodal dissection in PTC, 29 patients with a preoperative diagnosis of PTC and no clinical or ultrasonographic evidence of nodal involvement underwent cervicotomy and exposure of the thyroid gland, followed by Patent Blue V dye injection into the thyroid nodule. Total thyroidectomy was subsequently performed, resecting the lymph nodes at levels III, IV, VI and VII. The thyroid, SLN and the other lymph nodes were snap-frozen and submitted for both intra-operative and subsequent definitive pathological evaluation. Intra-operative lymphatic mapping located the SLN in 22/29 patients (75.9%) and the SLN revealed neoplastic involvement in 4/22 (18.2%); other lymph nodes were also positive in 2 cases. In the 18 patients whose SLNs were not metastatic, the other nodes were also disease-free. The SLN technique thus seems helpful in avoiding unnecessary lymph node dissection in PTC without spread to the SLN.

Adult↗
PubMedDate not supplied

Sentinel node mapping identifies vaccine-draining lymph nodes with tumor-specific immunological activity.

BACKGROUND: Adoptive immunotherapy (AIT) with 4T07-IL2 vaccine-draining lymph node (DLN) cells induced regression of established 4T07 mammary carcinomas, but contralateral non-DLN were inactive. These experiments were performed to determine whether mapping with isosulfan blue (IB), as described for identification of sentinel nodes, would identify vaccine-DLN with antitumor activity. METHODS: Ten days after vaccination with 4T07-IL-2, .1 ml of 1% IB was injected into the vaccination site (footpads or flanks). After 3 minutes, mice were euthanized, and the blue-stained nodes were collected. With flank vaccination, IB identified both an inguinal and an axillary node. We also collected DLNs blindly in mice not receiving IB dye. DLN cell suspensions were then activated with bryostatin 1, ionomycin, and IL-2, expanded in culture, and adoptively transferred to mice bearing established 4T07 flank tumors. RESULTS: Complete tumor regression occurred in nearly all mice treated with popliteal or inguinal DLNs collected with or without IB. IB-stained axillary DLNs cured 100% of tumor-bearing mice, whereas none of the mice treated with blindly collected axillary DLNs were cured. CONCLUSION: We have shown that IB identifies immunologically active DLNs, does not interfere with expansion of lymphocytes in vitro, and, more importantly, has no detrimental effect on the ability of lymphocytes to induce tumor regression in vivo. For axillary DLNs, use of IB mapping identified immunologically active lymph nodes that could not otherwise be found.

Animals↗
PubMed1995

The influence of patent blue V on pulse oximetry and haemoximetry.

Patent Blue V (PBV) is a blue dye solution which is used to visualize lymphatic vessels for surgical procedures. There has been some conflicting reports about the influence of this dye solution on pulse oximetry and haemoximetry, why we decided to: 1) produce a spectrum of PBV in plasma, 2) measure absorbances of full blood before and after addition of PBV and 3) to record the effect on the pulse oximeter and haemoximeter of an intradermal injection of PBV to a patient. Computer analysis of the blood gas measurements were carried out by the Oxygen Status Algorithm (OSA). The spectrum of PBV demonstrated a peak absorption at 640 nm and correspondingly the absorbances of the haemoximeter increased most significantly at 622 and 636 nm. These changes invalidated the determination of haemoglobin pigments, and the results should not be used. Computer analysis interpreted the measurements as a shift of the haemoglobin oxygen binding curve.

Absorption↗
PubMed2001

Aberrant drainage and missed micrometastases: the value of lymphatic mapping and focused analysis of sentinel lymph nodes in gastrointestinal neoplasms.

Lymph node analysis is essential for staging gastrointestinal (GI) neoplasms. Our group has conducted several studies of intraoperative lymphatic mapping and sentinel lymphadenectomy (LM/SL) for the staging of GI neoplasms. LM is performed following injection of 0.5-1 ml of isosulfan blue dye, and blue-stained sentinel lymph nodes (SLNs) are analyzed by hematoxylin and eosin (H&E) staining, multiple sectioning, and cytokeratin immunohistochemistry. In feasibility trials, LM identified at least one SLN in 121 of 126 patients. Of the 58 cases with nodal metastasis, 50 (89%) had at least one positive SLN and 24 (42%) had nodal metastasis only in the SLN. In 25 cases, tumor deposits were identified by multiple sectioning (n = 8) or immunohistochemistry (n = 17) only. In 10 cases (8%), LM identified aberrant lymphatic drainage that altered the extent of the lymphadenectomy. Our cumulative experience indicates that focused analysis of the SLNs draining GI neoplasms can increase the detection of micrometastases and may improve selection of patients for adjuvant treatment.

Adenocarcinoma↗
PubMed1980

Nonspecific ("pseudo-plasmal") dye-binding in the Feulgen nuclear stain and its blocking by azocarmin G.

In Feulgen nuclear staining nonspecific dye-binding due to the "pseudo-plasmal reaction" is intensified in isolated cells with intact cytoplasm, and cannot be eliminated by the post-irradiation method. Fluorescence intensity in the cytoplasm sometimes exceeds that of specific nuclear fluorescence, especially in brain and heart muscle cells, and it was almost impossible to perform cytofluorometric DNA quantification on such specimens. Various kinds of aldehyde-blocking agents such as sodium borohydride, 2,4-dinitrophenylhydrazine, aniline, and sodium pyrosulfite were effective in reducing the "pseudo-plasmal reaction". But the blocking effects were not complete because of additional release of reactive aldehyde groups during subsequent Feulgen hydrolysis. Acidic azocarmin G produced a complete block of all "pseudo-plasmal reaction" in acriflavine-Feulgen nuclear staining, allowing accurate DNA-cytofluorometry to be carried out.

Acriflavine↗
PubMed2003

Determination of residues of malachite green in aquatic animals.

Residues of malachite green (MG) were extracted from homogenized animal tissues with a mixture of McIlvaine buffer (pH 3.0)-acetonitrile, and purified over an aromatic sulfonic acid solid-phase extraction column followed by HPLC or LC-ESI-MS-MS analysis. Ascorbic acid and N,N,N',N'-tetramethyl-1,4-phenylenediamine dihydrochloride were added to reduce de-methylation of the dye. Responses were recorded at 620 nm (HPLC) or by multiple-reaction-monitoring (LC-MS-MS) after post-column oxidation using PbO(2). MG and its primary metabolite leuco-malachite green (LMG) were successfully determined at 2.5-2000 microg/kg in catfish, eel, rainbow trout, salmon, tropical prawns and turbot, with a limit of detection at 1 microg/kg (HPLC) and 0.2 microg/kg (LC-MS-MS) for both MG and LMG. Recoveries for LMG were between 86+/-15% (prawn) and 105+/-14% (eel). Freeze-thawing cycles, and storage at 4 degrees C and -20 degrees C affected the recovery of both MG and LMG. Analyses of eel, trout and (processed) salmon field samples collected at local retailers, fish-market and -shops demonstrated trace levels of MG-residues.

Animals↗
PubMed2001

Allergic reactions to isosulfan blue during sentinel node biopsy--a common event.

BACKGROUND: Sentinel lymph node (SLN) dissection in the management of high-risk melanoma and other cancers, such as breast cancer, has recently increased in use. The procedure identifies an SLN by intradermal or intraparenchymal injection of an isosulfan blue dye, a radiocolloid, or both around the primary malignancy. METHODS: At the time of selective SLN mapping, 3 to 5 mL of isosulfan blue was injected either intradermally or intraparenchymally around the primary malignancy. From October 1997 to May 2000, 267 patients underwent intraoperative lymphatic mapping with the use of both isosulfan 1% blue dye and radiocolloid injection. Five cases with adverse reactions to isosulfan blue were reviewed. RESULTS: We report 2 cases of anaphylaxis and 3 cases of "blue hives" after injection with isosulfan blue of 267 patients who had intraoperative lymphatic mapping by the procedure described above. The 2 patients with anaphylaxis experienced cardiovascular collapse, erythema, perioral edema, urticaria, and uvular edema. The blue hives in 3 patients resolved and transformed to blue patches during the course of the procedures. CONCLUSIONS: The incidence of allergic reactions in our series was 2.0%. As physicians expand the role of SLN mapping, they should consider the use of histamine blockers as prophylaxis and have emergency treatment readily available to treat the life- threatening complication of anaphylactic reaction.

Adult↗
PubMed2004

Sentinel lymph node biopsy in breast cancer: results of intradermal periareolar tracer injection and follow-up of sentinel lymph node-negative patients.

Several methods are in use for identification of the sentinel lymph node (SLN) in breast cancer. We set up the hypothesis that intradermal (i.d.) infra-areolar injection of technetium-99m in combination with i.d. injection of patent blue dye lateral to the areola can identify the same lymph node as peritumoral injection, regardless of the location of the tumour. Each of 50 patients with breast cancer (group I) received an i.d. injection of technetium-99m 1cm caudal to the areola. After induction, blue dye was injected intradermally 1cm lateral to the areola. These patients underwent axillary dissection regardless of their lymph node status. The SLN was identified in 96% of these patients. One of them had axillary lymph node metastases even though the SLN was negative (6%). Further 82 patients (group II) underwent SLN identification and removal without further axillary dissection. The duration of mean follow-up for these patients was 28 months (16-39 months). One patient developed axillary recurrence (1%) 24 months after the initial operation. Intradermal periareolar tracer injection is an accurate method of locating the sentinel node. Long-term follow-up of patients who had negative sentinel nodes and did not undergo axillary dissection revealed a low axillary recurrence rate.

Adult↗
PubMed2000

Evolution of sentinel lymph node biopsy for melanoma at a National Cancer Institute-designated cancer center.

BACKGROUND: Sentinel lymph node biopsy (SLNB) has rapidly evolved into the standard of care for clinically node-negative melanoma. Since adopting sentinel lymph node (SLN) technology in 1993, we have periodically reviewed our institution's results and made several modifications. METHODS: From January 1993 to December 1998, 182 patients with clinically node-negative primary cutaneous melanoma underwent SLNB. Charts were retrospectively reviewed and assessed for the technique for the identification of the SLN, the pathologic analysis, and the use of intraoperative frozen section. RESULTS: The accuracy of SLN identification improved from 91% to 100% with the combination of isosulfan blue dye and radiolabeled colloid over isosulfan blue dye alone. Routine versus selective lymphoscintigraphy identified 7 in-transit SLNs and increased detection of dual nodal basin drainage (15%-27%). Identification of micrometastases in the SLN increased from 14% to 24% after a modification of pathologic evaluation. The positive SLN was the only involved node in most patients (80%). Intraoperative frozen section had a sensitivity of 58% and was of benefit in only 13 of 124 patients (10%). CONCLUSIONS: Several modifications to the identification of the SLNs and the detection of metastatic melanoma have improved our outcome with SLNB. A careful, periodic review of results to identify areas for improvement at each institution is crucial to the success of SLNB for melanoma.

Adult↗
PubMed2002

Assessment of acrosomal status in rat spermatozoa: studies on carbohydrate and non-carbohydrate agonists.

In the mouse and several other species, including man, capacitated acrosome-intact spermatozoa interact with natural [soluble zona pellucida (ZP) and progesterone (P4)] and synthetic [neoglycoproteins (ngps) and calcium (Ca(2+)) ionophore] agonists, prior to the initiation of a Ca(2+)-dependent signal transduction cascade. The net result is the fusion of the sperm plasma membrane overlying the outer acrosomal membrane at multiple sites and exocytosis of acrosomal contents [i.e., induction of the acrosome reaction (AR)]. This step is believed to be a prerequisite that enables the acrosome-reacted spermatozoon to penetrate the ZP and fertilize the egg. Although the rat is one of the most commonly used laboratory animals, very little is known about the chemical nature of agonists that induce the AR in this species. The lack of this information is primarily due to the fact that the rat sperm acrosome is a relatively thin structure. Thus, it is difficult to assess the status of the sperm acrosome in this species. In this report, we describe the use of a Coomassie brilliant blue dye staining procedure to assess the status of the rat sperm acrosome by light microscopy. The procedure is highly reproducible and has allowed us to determine the effects of carbohydrate (ngps and mouse ZP) and noncarbohydrate (P4 and Ca(2+) ionophore) agonists on capacitated spermatozoa. In addition, we have used a pharmacological approach to examine the functional significance of calmodulin (CaM), a Ca(2+)-binding protein, in induction of the AR in spermatozoa. Data presented in this report demonstrate that several ngps, solubilized mZP, P4, and Ca(2+) ionophores induce the AR in rat spermatozoa. Furthermore, we demonstrate that, whereas CaM antagonists blocked P4-induced AR, most of the inhibitors used had no significant effect on the Ca(2+) ionophore-induced (nonphysiological) AR.

Acrosome Reaction↗
PubMed1996

Microleakage of direct and indirect inlay/onlay systems.

The purpose of this in vitro study was to evaluate the microleakage of direct and indirect inlay/onlay systems. Two of the groups investigated have indirect applications only and consist of microfine composites. The other two groups of inlay/onlay systems have both direct and indirect applications and are hybrid composites. Standardized MOD preparations were cut in 60 extracted human premolars. One gingival margin was placed in enamel above the cementoenamel junction (CEJ) and the other was placed in dentine below the CEJ. The inlays were made and luted according to each manufacturer's recommendation. The restored teeth were thermocycled and immersed in a basic fuchsin solution for 24 h. Following the mesiodistal sectioning of the specimens, extent of dye penetration at the restoration-glass ionomer cement and tooth-glass ionomer cement interfaces were scored using a stereomicroscope. Both direct and indirect inlays showed substantial leakage at gingival-dentine margins. However, there was only superficial leakage at enamel margins. Comparing the overall indirect inlay systems, EOS inlay leaked the most (P < 0.05). Indirect Brilliant Dentin inlays showed a significantly smaller amount of leakage than the other inlay systems. SR-Isosit and Estilux Posterior C VS inlays provided a favourable marginal seal. In the case of Brilliant Dentin and Estilux Posterior C VS, indirect inlays leaked more than direct inlays. Generally, no significant difference was observed between the interfaces evaluated in microleakage performance.

Acrylic Resins↗
PubMedDate not supplied

[Identification of sentinel lymph node with patent blue V in patients with cutaneous melanoma].

Regional lymphadenectomy in patients with cutaneous malignant melanoma in stages I-U AJC/UICC has not demonstrated improvement either in loco-regional control or in survival. The results of the lymphatic mapping technique have demonstrated that on identifying the sentinel node, the group of patients with microscopic nodal metastases can be selected for regional treatment. We performed the intraoperative lymphatic mapping technique with 1 ml of blue patent V by intradermal injection just around the primary melanoma; after injecting the dye we incised the nodal skin region, identifying the sentinel node by its blue color. The histologic examination, by frozen section of this node, determines whether or not to proceed with a formal lymphadenectomy. We calculated the sensitivity of the dye for the identification of the sentinel node and the Pearson's test was performed between the intraoperative histologic study and the definite pathological result. Thirty one lymphatic mappings were performed in 29 patients with stages I-II malignant cutaneous melanoma. The sentinel node was found in 26/31 explored lymphatic zones, and in 5, it was not found, sensitivity of 84%; 23/26 sentinel nodes were negative and only 3/26 were metastatic in frozen section. The Pearson's test result was 0.78 with a predictive value of 92%. The blue patent V intraoperative lymphatic mapping technique in patients with malignant cutaneous melanoma provided a great sensitivity to identify the sentinel node and allows the identification of patients with high risk of microscopic nodal metastases.

Adult↗