Sentinel lymph node mapping with indocyanine green compared to blue dye tracer in gynecologic malignancies-A single center experience of 218 patients.

BACKGROUND AND OBJECTIVES: To compare the detection rates of sentinel lymph nodes after converting the tracer technique from blue dye to indocyanine green (ICG). METHODS: Patients with uterine or cervical cancer were enrolled for sentinel lymph node (SLN) dissection. A total of 109 consecutive patients were analyzed and compared to a historical cohort of 109 consecutive patients with the sentinel blue dye technique. SLNs were analyzed by ultrastaging. RESULTS: The bilateral mapping rate of sentinel nodes was significantly higher with the ICG (78%; n = 85) compared to the blue dye tracer (61%; n = 67; p = .006). Neither the mean number of SLN nor the rate of low volume metastases showed significant differences between both cohorts. In the subgroup of endometrial cancer patients, the number of systematic lymph node dissection (LND) was significantly lower in the ICG cohort compared to the blue dye cohort (9% vs. 28%, p = .001). CONCLUSIONS: ICG improved the detection rate of pelvic SLN compared to blue dye and may be considered as the superior technique. In clinical practice, the rate of systematic LND further decreased after incorporating SLN mapping with ICG. Reliable safety data are still pending.

The Bio-Safety Concerns of Three Domestic Temporary Hair Dye Molecules: Fuchsin Basic, Victoria Blue B and Basic Red 2.

Hair-coloring products include permanent, semi-permanent and temporary dyes that vary by chemical formulation and are distinguished mainly by how long they last. Domestic temporary hair dyes, such as fuchsin basic, basic red 2 and Victoria blue B, are especially popular because of their cheapness and facile applications. Despite numerous studies on the relationship between permanent hair dyes and disease, there are few studies addressing whether these domestic temporary hair dyes are associated with an increased cancer risk. Herein, to ascertain the bio-safety of these temporary hair dyes, we comparatively studied their percutaneous absorption, hemolytic effect and cytotoxic effects in this paper. Furthermore, to better understand the risk of these dyes after penetrating the skin, experimental and theoretical studies were carried out examining the interactions between the dyes and serum albumins as well as calf thymus (CT)-DNA. The results showed that these domestic temporary hair dyes are cytotoxic with regard to human red blood cells and NIH/3T3 cell lines, due to intense interactions with bovine serum albumin (BSA)/DNA. We conclude that the temporary hair dyes may have risk to human health, and those who use them should be aware of their potential toxic effects.

Efficient decolourization of malachite green with biosynthesized iron oxide nanoparticles loaded carbonated hydroxyapatite as a reusable heterogeneous Fenton-like catalyst.

In this study, iron oxide nanoparticles (IO-NPs) with a mean diameter of 102.85 nm were firstly synthesized via a facile green route using Ulva spp. aqueous extract as a bioreductant agent. Then, IO-NPs were loaded into carbonated hydroxyapatite (c-Hap) and the final product was named as the iron oxide nanoparticles loaded carbonated hydroxyapatite (IO-NPs-Lc-Hap). Subsequently, IO-NPs-Lc-Hap was characterized by FT-IR, SEM, XRD and EDX analysis methods. MG colour removal efficiencies of Ulva spp., Hap, IO-NPs and IO-NPs-Lc-Hap materials were also evaluated by adsorption and/or Fenton-like reaction methods. IO-NPs-Lc-Hap with the highest decolourization capacity was chosen as a heterogeneous Fenton-like catalyst for Malachite Green (MG). For Fenton-like decolourization of MG, the optimum H2O2 concentration, initial dye concentration and catalyst concentration were determined to be 30 mM, 100 mg/L and 1.0 g/L, respectively. At these optimum conditions, 100% decolourization efficiency and 33.3% COD removal were obtained. On the other hand, 94% decolourization efficiency and 42% COD removal were achieved for the real textile wastewater at the obtained optimum conditions. The experimental decolourization reaction rate for MG was determined as -rd = 0.0779 [(mg dye0.3) (g cat-0.3) (min-1)] × qt0.7. Also, the catalyst had high decolourization efficiencies at the end of six sequence usages.

Axillary reverse mapping with indocyanine green or isosulfan blue demonstrate similar crossover rates to radiotracer identified sentinel nodes.

BACKGROUND: Sentinel lymph node (SLN) resection is imperative for breast cancer staging. Axillary reverse mapping (ARM) can preserve arm draining nodes and lymphatics during surgery. ARM is generally performed with isosulfan blue (ISB), restricting its use for concurrent SLN biopsy. Indocyanine green (ICG) could serve as an alternative to ISB for ARM procedures. METHODS: SLN mapping and biopsy was performed via periareolar injection of 99 technetium-sulfur colloid (99m TcSc, TSC). ISB and ICG were injected in the upper arm. Blue-stained lymphatics or nodes were visualized in the axilla; ICG was identified using the SPY Elite® system. RESULTS: Twenty-three patients underwent SLN biopsy with or without axillary node dissection and ARM procedures. Twenty of these patients had at least one hot node; 12 patients had SLNs that were only hot, 6 hot/blue/fluorescent, and 2 hot/fluorescent. Overall, crossover of ARM agents with SLNs occurred in 8 cases. Inspection of the axillary cavity after SLN biopsy revealed fluorescent lymphatics and nodes remaining in 14 and 7 patients, respectively. Blue lymphatics and blue nodes were detected in fewer cases. CONCLUSION: Nearly one-third of patients showed crossover between breast and arm draining nodes, which provides insight as to why some patients develop lymphedema symptoms after SLN biopsy. ICG and ISB identify similar numbers of SLNs. As such ICG could substitute for ISB in ARM procedures.

Study on enhanced lymphatic tracing of isosulfan blue injection by influence of osmotic pressure on lymphatic exposure.

Isosulfan blue (IB) is being used as a lymphatic tracer has been approved by the FDA in 1981. This study aimed at improving lymphatic exposure of IB injection by osmotic pressure regulation to achieve step-by step lymphatic tracing. First, IB injection with appropriate osmotic pressure, stability, and suitable pH was prepared. Next, the lymphatic tracing ability of different osmotic pressure was studied to determine the blue-stained state of IB in three-level lymph nodes after subcutaneous administration. Furthermore, pharmacokinetics of lymphatic drainage, lymph node uptake, and plasma concentration was investigate to explore the improving law of the lymphatic tracing by osmotic pressure, and combined with tissue irritation to determine the optimal osmotic pressure. At last, the tissue distribution in mice of IB injection which had the property of optimal osmotic pressure was investigated. The results showed that increasing osmotic pressure could significantly reduce injection site retention and increase IB concentration of lymph node. The lymph nodes could be obviously blue-stained by IB injection which had 938 mmol/kg osmotic pressure and would not cause inflammatory reaction and blood exposure. The tissue distribution study suggested that IB injection which had 938 mmol/kg osmotic pressure was mainly distributed into gallbladder and duodenum that verified the reports that 90% IB was excreted through the feces through biliary excretion. In conclusion, this study provides the basic study to improve lymphatic exposure of IB injection by regulate the osmotic pressure and have the potential to be the helpful guidance for the elective lymph node dissection.

An evaluation of two heavier-than-water internal limiting membrane-specific dyes during macular hole surgery.

PURPOSE: To evaluate the staining characteristics and effect on internal limiting membrane (ILM) histology of two heavier-than-water ILM-specific dyes during macular hole surgery: acid violet 17 combined with 5 % mannitol (AV17-M) and brilliant blue G with 4 % polyethylene glycol (BBG-P). METHODS: Single-centre observational comparative cohort study. The ILM of consecutive patients undergoing surgery for idiopathic macular hole were stained with BBG-P and AV17-M for 10 s each. The ILMs were retrieved and examined with electron microscopy. The extent of retinal and vitreous side debris was scored. Surgical videos were used to assess the staining contrast effect by measuring the Euclidean distance in the CIELAB colour space between stained and unstained retinas after peeling. RESULTS: 51 consecutive patients were studied with 25 in the AV17-M group and 26 in the BBG-P group. The mean age was 71 years with no significant difference between the groups. The amount of retinal side tissue was greater on the BBG-P-stained ILMs compared to the AV17-M-stained ILMs (30.2 versus 19.6 %, p < 0.001). There was a difference in the CIELAB colour space separation distance between stained and peeled retinas (5.89 versus 3.97, p = 0.01) in favour of BBG-P. Visual outcomes between the two groups were similar (logMAR visual acuity 0.40 versus 0.38, p = 0.74). CONCLUSION: Both stains were successfully used to peel the ILM with comparable outcomes. AV17-M resulted in less retinal debris than BBG-P, suggesting an altered and potentially beneficial ILM cleavage plane from the retina but with lowered staining contrast than BBG-PEG.

Shaving effects on percutaneous penetration: clinical implications.

CONTEXT: Human/animal shaving biology. OBJECTIVE: To assess the effect of shaving on percutaneous penetration and skin function. METHODS: We screened 500+publications in Pub Med, Scopus, Cochrane Library and pertinent journals out of which only 17 were deemed relevant. Terms for searches included shaving and skin, percutaneous penetration and shaving, skin absorption and shaving, absorption of dyes and shaving, skin penetration, effects of shaving and absorption, shave and dyes, axillary shaving and stratum corneum, shaving and breast cancer, shaving and infections, etc. RESULT: Shaving appears to have an exaggerated effect on percutaneous absorption; however, some studies do not support this evidence. CONCLUSION: Shaving enhances percutaneous penetration of some chemicals; however this effect is species and chemical specific. Further investigations of chemicals of varying physio-chemical properties are mandated before a generalized theory can be promulgated.

Quantitative analysis of vitreous and plasma concentrations of brilliant blue g after use as a surgical adjuvant in chromovitrectomy.

PURPOSE: To measure the concentration of brilliant blue G (BBG) in vitreous and plasma after use as a surgical adjuvant for staining and peeling of the internal limiting membrane to determine potential systemic adverse effects. METHODS: This study was designed as a prospective, interventional, clinical, case series. Five eyes from five patients with macular hole or epiretinal membrane underwent BBG-assisted internal limiting membrane and epiretinal membrane removal. The vitreous samples were obtained and stored at the end of surgery in all five cases. The plasma specimens were extracted and stored at the end of the operation, after 4 hours, and after 7 days post operation. For BBG analysis of plasma and vitreous, high-performance liquid chromatography coupled with tandem mass spectrometric detection was used. RESULTS: Brilliant blue G was not detected in plasma from all five cases at the three points of measurement. The mean vitreous BBG concentration was 34.5 ± 23.7 ng/mL (range, 11.3-70.9 ng/mL). Postoperative progress was good, and adverse effects were not observed in any of the five cases. CONCLUSION: Brilliant blue G, which remained at low levels in the vitreous cavity, was not found in the systemic blood flow after the operation. Thus, any adverse effects of systemic BBG would be avoided.

Dye-assisted lymphatic-sparing laparoscopic varicocelectomy in children.

INTRODUCTION: The ideal method for varicocelectomy in children remains controversial. We present our experience with dye-assisted lymphatic-sparing laparoscopic varicocelectomy (LSLV) in children, which overcomes the limitations of previously described techniques. MATERIALS AND METHODS: Twenty-five consecutive LSLVs were performed on children with a mean age of 15 years over a 4-year period. Varicocele grade was 3 in 21 cases and grade 2 in 4. Indications for intervention were hypotrophy in 12, pain in 11 cases and family preference in 2. A scrotal injection of lymphatic dye was utilized to spare at least one lymphatic and the remaining spermatic vessels were divided. RESULTS: Lymphatic sparing was accomplished in all cases. Operative time varied from 30 to 140 min (mean 85 ± 26). No perioperative complications were noted. On average follow-up of 13 months a residual varicocele was noted in 2 cases, with no hydrocele and resolution of pain. Mean testicular volume difference diminished from 33% pre to 18% postoperatively. CONCLUSION: This multi-surgeon experience demonstrates that dye-assisted LSLV is easily accomplished with promising results. It appears that preservation of a single spermatic lymphatic vessel is sufficient, although in some cases a second dye injection is required to visualize the lymphatics.

Staining internal limiting membrane with a mixture of BBG and sodium hyaluronate.

AIMS: To evaluate the intraoperative applicability and safety of a mixture of brilliant blue G and sodium hyaluronate (visco-BBG) for staining the inner limiting membrane (ILM). METHODS: A retrospective consecutive case series. Seventy-four eyes that had undergone ILM peeling were studied. During vitrectomy, ILM peeling with visco-BBG (visco-BBG group) was performed on 40 eyes; 12 with a macular hole (MH), 26 with an epimacular membrane (ERM) and 2 with a retinal detachment due to a MH (MHRD). ILM peeling with BBG dissolved in balanced salt solution (BSS-BBG group) was performed on 34 eyes; 9 with a MH, 23 with an ERM and 2 with a MHRD. The main outcome measures were the distribution of the dye within the vitreous cavity and the retinal sensitivity in the MH patients of the two groups by microperimetry. RESULTS: The visco-BBG was injected over the retina where the ILM was intended to be peeled, and it stained the ILM in all cases. It did not disperse throughout the vitreous cavity or into the subretinal space. The BSS-BBG dispersed throughout the vitreous cavity, and its distribution was difficult to control. The two solutions did not stain the epiretinal membranes or any residual posterior hyaloid membrane. The difference in the retinal sensitivity between the two patients with MH of two groups was not significant. No complications were found in the visco-BBG group, although an accidental retinal perforation was found in one eye of the BSS-BBG group. Transmission electron microscopy confirmed that the membrane peeled was the ILM. CONCLUSIONS: Visco-BBG can be a useful method to assist macular surgery and can overcome some of the disadvantages of conventional BBG solutions dissolved in BSS.

"Brilliant Blue G" and "Membrane Blue Dual" assisted vitrectomy for macular hole.

The aim of this study is to evaluate vital dyes "Brilliant Blue G" (BBG) and "Membrane Blue Dual" (MBD) for intraoperative staining of the inner limiting membrane (ILM) during vitrectomy for macular hole (MH). Retrospective, comparative case series on 18 eyes with macular holes who underwent "23 and 25 gauge" pars plana vitrectomy. Main outcome measurements were staining intensity and characteristics, visual acuity, visual field, OCT measurements and complications over a period of 6 months. With the help of BBG and MBD successfully was removed complete ILM in 14 eyes. Postoperative visual acuity was improved in 12 patients, unchanged in 2 patients and worse in 4 patients. Central retinal thickness showed significant postoperative reduction with closure of macular hole. OCT values range were from -10 to -250 microm. No visual field defects and no adverse effects were found. BBG and MBD successfully identificate internal limiting membrane during vitrectomy for MH. Good anatomical and functional results are achieved with the use of both vital dyes.

Thoracic duct cannulation without thoracotomy in sheep: a method for accessing efferent lymph from the lung.

We have developed and validated a novel method to access efferent lymph draining the lung and gut of sheep. In this model, efferent lymph derived from the lung could be collected via cannulation of the thoracic duct just prior the thoracic duct-jugular vein junction. The thoracic duct was accessed in the neck region without needing to broach the thoracic cavity, thus avoiding extensive tissue damage to the animal and need for ventilation during surgery. In addition, this surgical approach allows for a second cannulation of an adjacent lymphatic draining the head/neck region, providing for an 'in-built' internal control with which to compare lymph parameters. To test the verity of cannulation procedure, a test protein ovalbumin (OVA) was infused into the left and right lungs via bronchoscopy. We found that OVA was recovered almost exclusively in the lymph draining the lungs compared to the lymph draining the head/neck where it was essentially non-existent. The method described here will be invaluable for optimizing intra-lung delivery of drugs or vaccines. In addition, access to lymph will also allow for analysis of immune responses to infections originating at this site.

Targeted delivery of pharmacological agents into rat dorsal root ganglion.

We sought an optimal method for targeted delivery into dorsal root ganglia (DRGs) for experimental studies, in terms of precision of delivery and avoidance of behavioral disturbances. We examined three approaches for injection into rat DRGs: percutaneous injection without surgical exposure, injection after deep exposure, and injection following deep exposure and partial laminectomy. Coomassie blue and Fast Blue were injected into DRGs for validation. At necropsy, the spread of Coomassie blue and Fast Blue was investigated under stereomicroscope and fluorescent microscope, respectively. We found that percutaneous approach did not provide any successful DRG injections. Deep exposure prior to intraganglionic injection provided variable results, but intraganglionic injection after deep exposure plus partial laminectomy was successful in 100% of attempts. Our subsequent skeletal analysis showed that the anatomical location of DRG is not compatible with successful DRG injection without surgical exposure. Neither of the methods using surgical exposure caused behavioral disturbances. Based on these results we conclude that partial laminectomy offers the most precise method of injecting DRG and does not produce behavioral evidence of nerve damage. Intraganglionic injection after deep exposure alone is less predictable, while percutaneous approaches only allow injection in the peripheral nerve.

Use of avidin/biotin-liposome system for enhanced peritoneal drug delivery in an ovarian cancer model.

The goal of this study was to determine the distribution of the avidin/biotin-liposome system in an ovarian cancer xenograft model. Optimal avidin/biotin-liposome injection sequence with enhanced liposome accumulation to the peritoneum was determined. Two weeks after NIH:OVCAR-3 cell inoculation, rats were divided into three groups. Group 1 (B-A) (n=4), received an intraperitoneal injection of (99m)Tc-blue-biotin-liposomes 30 min before an intraperitoneal injection of avidin. Group 2 (A-B) (n=4), received an intraperitoneal injection of avidin 30 min before an intraperitoneal injection of (99m)Tc-blue-biotin-liposomes. Group 3 (A-B 2h) (n=5), received an intraperitoneal injection of avidin 2h before an intraperitoneal injection of (99m)Tc-blue-biotin-liposomes. Three additional non-tumor nude rats served as controls in each group, and were subjected to the same injection sequences. Scintigraphic imaging commenced at various times post (99m)Tc-blue-biotin-liposome injection. After imaging, rats were euthanized at 23 h post-liposome injection for tissue biodistribution. Images showed no apparent difference in liposome distribution between control and tumor animals. Regional uptake analysis at 4h for tumor rats showed significantly higher lymphatic channel uptake in the A-B 2h group (p<0.05) and a trend of increased peritoneal uptake in A-B group. By 22 h, peritoneal and lymphatic channel uptake was similar for all groups. At necropsy, most activity was found in blue-stained omentum, diaphragm, mediastinal and abdominal nodes. Bowel activity was minimal. These results correlate with previous normal rat studies, and demonstrate potential use of this avidin/biotin-liposome system for prolonging drug delivery to the peritoneal cavity and associating lymph nodes in this ovarian cancer xenograft model.

Patent blue: a novel vital dye in vitreoretinal surgery.

PURPOSE: To determine a novel vital dye (patent blue; Blueron) for vitreoretinal surgery in a prospective consecutive case series. METHODS: Five patients with either idiopathic epiretinal membrane (ERM; n = 2), proliferative vitreoretinopathy (n = 2), or a macular hole (n = 1) underwent a three-port pars plana vitrectomy. Patent blue assisted staining of the retinal surface followed by a consecutive peeling of the ERM (n = 4) or of the internal limiting membrane (ILM; n = 1) was performed. The main outcome measures were quality of intraoperative visualization of preretinal structures and postoperative visual acuity. RESULTS: The dye induced a moderate staining (++) of the ERM and a mild staining (+) of the ILM. Complete ERM and ILM removal was successfully achieved in all cases. A mean visual improvement of three Snellen lines was observed 6 months postoperatively. No visual field defects or visible retinal pigment epithelial changes were present 6 months postoperatively. CONCLUSION: Patent blue, a novel dye for intraocular applications, may be added as an alternative dye in chromovitrectomy.

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.

Biotransformation of malachite green by Saccharomyces cerevisiae MTCC 463.

In recent years, use of microbial biomass for decolourization of textile industry wastewater is becoming a promising alternative in which some bacteria and fungi are used to replace present treatment processes. Saccharomyces cerevisiae MTCC 463 decolourized the triphenylmethane dyes (malachite green, cotton blue, methyl violet and crystal violet) by biosorption, showing different decolourization patterns. However, malachite green decolourized by biosorption at the initial stage and further biodegradation occurred, about 85% in plain distilled water within 7 h, and about 95.5% in 5% glucose medium within 4 h, under aerobic conditions and at room temperature. Decolourization of malachite green depends on various conditions, such as concentration of dye, concentration of cells, composition of medium and agitation. HPLC, UV-VIS, FTIR and TLC analysis of samples extracted with ethyl acetate from decolourized culture flasks confirmed the biodegradation of malachite green into several metabolites. A study of the enzymes responsible for the biodegradation of malachite green in the control and cells obtained after decolourization showed the activities of laccase, lignin peroxidase, NADH-DCIP reductase, malachite green reductase and aminopyrine N-demethylase in control cells. A significant increase in the activities of NADH-DCIP reductase and MG reductase was observed in the cells obtained after decolourization, indicating a major involvement of reductases in malachite green degradation.

A sheep cannulation model for evaluation of nasal vaccine delivery.

We have developed and validated a novel model to investigate the efficacy of nasal vaccine delivery. Based on lymphatic cannulation of the tracheal lymph trunk of sheep, the model allows collection of lymph draining from the Nasal Associated Lymphoid Tissue. The model is suitable for determining both the amount of material that is absorbed into the lymphatic system, following intra-nasal delivery and the immune response that occurs following vaccination into the nasal area. The cell populations that track in this duct were phenotyped and found to be similar to those previously reported to be present in efferent lymph draining from peripheral lymph nodes. Following intra-nasal spray, we demonstrated that the amount of material recovered in draining lymph is only a very small fraction of the total delivered. Nevertheless, intra-nasal spraying of a vaccine could activate local immune cells. The method described will be invaluable for optimising intra-nasal delivery systems by allowing a separate optimisation of vaccine uptake and immune responses induction.

Cutaneous distribution and localization of dyes affected by vehicles all with different lipophilicity.

The penetration of topically applied substances into the stratum corneum (SC) depends on several factors, e.g., the physicochemical properties of the vehicle used for application. The penetration of highly hydrophilic and lipophilic dyes into the skin was studied using a pure oil (o) or water (w) for the application compared to an o/w emulsion. The penetration and localization of both dyes, the lipophilic curcumin and the hydrophilic Patent blue V, was investigated in vivo using the method of tape stripping and microscopy. In addition, histological sections of biopsies, removed from porcine ear skin were studied using microscopy. Differences in the distribution and the localization of both dyes within the SC were observed. These differences depend on the physicochemical properties of both the vehicles and the dyes. The vehicle appears to affect, in particular, the pathways of penetration.