Blue Dye Method Research Articles

The value of contrast\u2010enhanced ultrasound in determining the location of sentinel lymph nodes in breast cancer

BackgroundThis study aimed to explore the sentinel lymph node (SLN) identification rate in breast cancer by subcutaneous and intradermal injection of ultrasound contrast agent in the mammary areola region, compared to the results achieved with methylene blue (MB).MethodsA total of 390 breast cancer patients with planned sentinel lymph node biopsy from our breast surgery department from July 2017 to February 2019 were enrolled. All patients were subjected to preoperative contrast-enhanced ultrasound (CEUS), that involved an intracutaneous injection of 1 mL ultrasonic contrast agent (UCA) at 3 and 6 o ‘clock, as well as a subcutaneous injection of 1 mL UCA at 9 and 12 o’clock. The enhanced lymph nodes along the enhanced lymphatic vessels from the mammary areola were traced. The number of enhanced lymph nodes were recorded, and an ultrasound-guided injection of 1:10 diluted carbon nanoparticles were used to mark all first site enhanced lymph nodes (i.e., SLNs). An intraoperative dye method (MB) was used to track the SLNs and the results were compared with the CEUS findings.ResultsAmong the 390 cases of breast cancer, enhanced SLNs were observed in 373 patients after an injection of UCA with an identification rate of 95.64 % (373/390), compared to the identification rate of 92.05 % (359/390) using the intraoperative MB. The difference between the two methods was statistically significant (P = 0.016). And among the 390 patients, a total of 808 enhanced lymph nodes were traced by preoperative CEUS, with a median of 2 (1,3). A total of 971 blue-stained lymph nodes were traced using the intraoperative MB, with a median of 2 (2,3), indicating a statistically significant difference (p < 0.001).ConclusionsIntradermal and subcutaneous injections of UCA in the mammary areola region may have clinical application value for the identification and localization of SLNs in breast cancer patients. The identification rate is higher than that of blue dye method, which can be used as a new tracer of sentinel lymph node biopsy and complement other staining methods to improve the success rate.

Study of Urinary Glycosaminoglycans among Essential Hypertensive Patients

Introduction: Essential hypertension is a systemic disease which affects endothelial basement membrane. Changes in Glycosaminoglycans (GAG) distribution pattern on glomerular basement membrane has been noted in hypertension. Hence it seems likely that an increase excretion of GAG levels may be an indicator of reduction in renal function. The qualitative or quantitative determination of urinary GAGs may be of value and could constitute a non invasive marker to assess the renal damage in essential hypertension. Aim: Differential analysis of urinary GAGs in essential hypertensive patients. Materials and Methods: This analytical case-control observational study which was conducted from November 2014 to June 2016 in Department of Biochemistry, SDM College of Medical Sciences and Hospital, Dharwad, Karnataka, India. The study group included 50 male patients with age group of 30-60 years, clinically diagnosed with essential hypertension. Control group included 50 healthy male individuals with age group of 30-60 years, blood donors visiting to hospital blood bank. Random urine sample was collected. Urine creatinine was estimated by Jaffe’s method, urine GAGs by Dimethyl methylene blue (DMMB) dye method and urine microalbumin by particle enhanced Turbidimetric inhibition immunoassay method. Correlation was tested with Spearman’s correlation coefficient. Level of significance was set for p-value &lt;0.05 with confidence interval of 95%. Results: In present study, urinary GAGs levels in essential hypertensive study group was 14.57±10.16 mg/dL and in control group was 10.09±6.04 mg/dL. Urinary GAGs levels in essential hypertensive group was significantly high (p-value=0.890) when compared with normotensive control group. But no statistical significant correlation was found between urine GAGs and urine microalbumin in hypertensive study group. Conclusion: Estimation of urine glycosaminoglycans in essential hypertensive patients is a simple, rapid and cost effective test which asseses the glomerular function. It can be used as one of the early marker for diagnosis of nephropathy before microalbuminuria sets in.

Angiopoietin-2 is released during anaphylactic hypotension in anesthetized and unanesthetized rats.

Angiopoietin (Angpt)-2, a permeability-increasing growth factor, is involved in vascular leakage of sepsis and acute lung injury, and could be released from endothelium in response to anaphylaxis-related secretagogues such as histamine and leukotrienes, or cytokines. However, roles of Angpt-2 in the hyperpermeability during systemic anaphylaxis are not known. Thus, we determined plasma levels of Angpt-2 and cytokines and vascular permeability during anaphylactic hypotension in unanesthetized rats. Anaphylaxis was induced by an intravenous injection of ovalbumin antigen. Mean arterial blood pressure (MBP) was measured, and hematocrit (Hct) and plasma levels of Angpt-2 and cytokines were assessed for 24 h after antigen injection. Separately, vascular permeability was measured in various organs using the Evans blue dye method, and Angpt-2 mRNA expression in liver was measured. After antigen injection, MBP decreased to the nadir at 6 min, and returned to baseline at 45 min, and Hct peaked at 20 min and thereafter progressively declined, suggesting that vascular leak and hypotension occurred within 20 min. Plasma Angpt-2 levels began to increase significantly at 1 h after antigen, reaching the peak 2.7-fold baseline at 6 h with a return to baseline at 24 h. Detected cytokines of IL-1α, IL-1β, IL-6, IL-10, and TNF-α peaked 1 or 2 h after antigen. Angpt-2 mRNA increased at 2 h and showed an increasing tendency at 6 h. Vascular permeability in bronchus, trachea, intestines, mesentery and skeletal muscle was increased at 10 min but not at 6 h after antigen. In addition, we confirmed using anesthetized rat anaphylaxis models that plasma Angpt-2 levels increased at 1 h after antigen. In conclusion, plasma Angpt-2 is elevated presumably due to increased cytokines and enhanced gene transcription during anaphylaxis in anesthetized and unanesthetized rats.

Sentinel lymph node biopsy after neoadjuvant treatment of breast cancer using blue dye, radioisotope, and indocyanine green: Prospective cohort study

BackgroundThe breast cancer treatment paradigm has shifted to neoadjuvant treatment. There are many advantages to neoadjuvant treatment, such as tumor downsizing, in vivo tumor biology testing, treating micrometastasis, and achieving complete pathological response (a surrogate marker for overall survival). However, in the post neoadjuvant settings, sentinel lymph node biopsy can be done using a dual staining technique to decrease the false-negative rate (FNR) and increase the detection rate. However, many hospitals are not equipped to use radioisotopes. Here we investigate the detection rate and accuracy of sentinel lymph node biopsy in post neoadjuvant treatment breast cancer, comparing radioisotope, isosulfan blue, and indocyanine green (ICG) approaches. Material and methodsThis prospective study includes breast cancer patients (T2–4, N1–2) who had received neoadjuvant treatment. Carcinomas were confirmed by tissue pathology. Patients who had previous surgical biopsy or surgery involving the axillary regions, and those with a history of allergy to ICG, isosulfan blue, or radioisotope were excluded from the study. ResultThe study was done between July 1, 2019 to March 31, 2020. The mean age of participants was 53 years. Fourteen (60.87%) were post-menopause, two (8.7%) were perimenopause, and seven (30.43%) were premenopause. The clinical-stage distribution of the participants was: 2A (8.7%), 2B (34.78%), 3A (43.48%), and 3B (13.04%). The primary tumor size was 4.82 ± 2.73 cm. The lymph node size was 1.8 ± 0.96 cm. The detection rates at the individual level were 95.23% with ICG, 85.71% with isosulfan blue, and 85.71% with a radioisotope. The detection rate increased up to 100% when the ICG and blue dye methods were combined. The FNRs of sentinel lymph node biopsy at the individual level were: 10% using ICG, 30% using isosulfan blue, and 40% using radioisotope. At the lymph node level, the detection rates were 93.22% using ICG, 81.78% using isosulfan blue, and 53.87% using a radioisotope. The FNRs of sentinel lymph node biopsy at the lymph node level were 19.05% with ICG, 21.43% with isosulfan blue, and 18.03% with a radioisotope. However, the FNR was less than 10% when ICG, isosulfan blue, and a radioisotope were combined. ConclusionWe can perform sentinel lymph node biopsy by combining blue dye with ICG as an optional modality and achieve a comparable outcome with combine radioisotope in locally advanced breast cancer after neoadjuvant treatment.

The Sensitivity and Specificity of Chicago Sky Blue (CSB) Dye in Comparisson with Potassium Hydroxide (KOH) Method for Superficial Dermatomycosis

Abstract: Superficial dermatomycosis is a skin, nail and hair infection caused by fungal pathogen. Based on the pathogen, this infection can be divided into dermatophytosis, pityriasis versicolor and superficial candidiasis. The rapid and proper diagnosis is necessary to determine the initial theraphy and prevent the treatment delay. Superficial dermatomycosis diagnosis can be performed using anamnesis, physical examination or supporting investigation. The routine investigation method commonly use Potassium Hydroxide (KOH) because the KOH method is easy to be performed, rapid, simple and affordable. Chicago Sky Blue (CSB) is a dye to give a better color contrast to the fungi so the fungi would be easier to be detected. Objectives of this research is to observe the sensitivity and specifity difference of Chicago Sky Blue (CSB) dye and Potassium Hydroxide (KOH) methods for Superficial Dermatomycosis. The research was performed using cross sectional design analitical obsevation with 30 research subjects. The subjects consist of 15 superficial dermatomycosis patients and 15 non-superficial dermatomycosis patients. The samples were taken from the patients lesion swabs. The samples were checked using KOH and CSB, then observed by the medical analyst. The superficial dermatomycosis samples consist of mostly dermathophytosis (53.33%), then pityriasis versicolor (26.67%) and superficial candidiasis (20%). The sensitivity and specificity of KOH were 86.67% and 100%, respectively. The sensitivity and specificity of CSB were 93.33% and 100%, respectively. The CSB dye method has a higher sensitivity than KOH. The fungal elements are nicely dyed and more easily detected using CSB dye. Keywords: KOH, Chicago Sky Blue, sensitivity, specificity, superficial dermatomycosis

The effect of low doses of doxorubicin on the rat glioma C6 cells in the context of the proteins involved in intercellular interactions

The aim of this investigation was to determine the effect of doxorubicin on F–actin rearrangement and β–catenin and cofilin–1 in a rat glioma C6 cell line in combination with changes in their morphology and ultrastructure. The experimental material constituted rat glioma C6 cell line. The cells were incubated with sublethal doses of doxorubicin in the concentration of 50, 100 and 200 nM. The blue trypan dye method was used to determine the number of dead cells. Morphological and ultrastructural changes in the cells were evaluated using light and transmission electron microscope, respectively. In order to determine the rearrangements and level of expression of F–actin, β–catenin and cofilin–1 they were analyzed using a fluorecence microscope. In turn, cell death and cell cycle were evaluated by Guava 6HT–2 L Cytometer. The performed experiments showed a dose–dependent decrease in the survival of C6 cells after treatment with doxorubicin. The analysis of cell death showed a dose–dependent increase in the population of apoptotic and necrotic cells. These results were confirmed by microscopy observation. The changes in morphology, ultrastructure, and rearrangements of F–actin, β–catenin and cofilin–1 were also observed. The results obtained in the study showed that sublethal concentrations of doxorubicin influenced the structure of F–actin and other proteins involved in cell–cell interactions. Moreover, mitotic catastrophe may preceding apoptosis, what suggest the cytotoxic effect of low dose of doxorubicin. Furthermore, our results confirmed the multi–dimensional mechanism of DOX action in tumor cells.

Contrast-enhanced ultrasonography and blue dye methods in detection of sentinel lymph nodes following neoadjuvant chemotherapy in initially node positive breast cancer.

Recent studies show that contrast-enhanced ultrasonography (CEUS) using SonoVue has the potential to improve the performance of sentinel lymph node biopsy (SLNB) in patients with early breast cancer. However, the evidence of SLNB using CEUS in patients converting from cN1 to cN0 after neoadjuvant chemotherapy (NAC) is lacking. The aim of this prospective study is to evaluate the feasibility of CEUS using SonoVue for the identification of sentinel lymph node (SLN) and the value of the combination of CEUS and blue dye (BD) for SLNB in patients converting from cN1 to cN0 following NAC. Patients with cytology-proven node positive breast cancer at the initial diagnosis (stage T1-T3N1M0) from January 2018 to January 2019, underwent NAC. Patients converting from cN1 to cN0 following NAC were enrolled and randomized into two groups for SLNB: the combination method group using CEUS and BD together, and the single BD method group. Then all patients underwent complete axillary lymph node dissection (ALND) and primary breast surgery. Compared with the final pathological results, the identification rate, sensitivity, specificity, accuracy, false negative rate, negative predictive value, positive predictive value were recorded and compared between two methods. A total of 400 patients with stage T1-T3N1M0 disease underwent NAC between January 2018 to January 2019, among which 134 (33.5%) patients had clinically negative node confirmed by imaging after NAC and randomized into two groups. Each group included 67 cases. In the combination method group, contrast-enhanced lymphatic vessels in 66 cases of 67 were clearly visualized by US soon after the periareolar injection of SonoVue and the SLNs were accurately localized. The identification rate of the combination method was 98.5%%, which was significantly higher than 83.6% (56/67) using the single BD method. The mean numbers of SLNs identified by the combination method was higher than that by the single BD method. Compared with pathological diagnosis, sensitivity, specificity, accuracy, the positive predictive value, the negative predictive value, and the FNR of the combingation method were 84.4%, 100%, 89.4%, 100%, 75%, and 15.6%, respectively. In contrast, sensitivity, specificity, accuracy, the positive predictive value, the negative predictive value, and the FNR using single blue dye were 73.9%, 100%, 89.3%, 100%, 84.6%, and 26.1%, respectively. The FNR using the combination method was significantly lower than that using single BD. Identification of SLNs in patients converting from cN1 to cN0 following NAC by CEUS is a technically feasible. The combination of CEUS and BD is more effective than BD alone for SLNB in patients converting from cN1 to cN0 following NAC.

Number of sentinel lymph nodes (SLN) removed in breast cancer (BC) patients undergoing sentinel lymph node biopsy (SLNB) in the province of Saskatchewan, Canada: Real-world experience.

e19240 Background: SLNB is considered standard of care for BC patient's undergoing surgery. This standard of care is based on several randomized, prospective clinical trials showing non inferiority of this procedure compared to full axillary lymph node dissection (FALND). Retrieval of less than 3 lymph nodes at the time of SLN biopsy has been shown to be associated with decrease in disease specific survival. In the NSABP B-32 study, removal of two SLN rather than one almost halved the false negative rate. SK has an area of 651,900 square kilometer, with a small population of 1,098,352. This population is widely distributed throughout the province with variable access to health care. This study provides us with a unique opportunity to see if data obtained in clinical trials can be replicated in community settings. Methods: After approval from SK Cancer Agency and research ethics board of University of SK. Female Patient’s with BC who underwent SLNB from Jan 2010 to Dec 2014, were identified using cancer registry. 529 charts were reviewed retrospectively. Exclusion criteria’s were Male BC, pathology other than invasive ductal or lobular, locally advanced or metastatic disease, T4 tumors, patients who received neo adjuvant therapy, patients who had declined or were unable to receive post-operative recommended adjuvant systemic or radiation therapy, patients with full axillary lymph node dissection. Data was analyzed using SAS 9.4 software. Results: Rate of 1, 2 and 3 or more SLN was 21, 35 and 44% respectively. The median number of lymph nodes retrieved were 2. 14% of patients had positive lymph nodes. 259/529 (48.9 %) of patients used both vital blue dye and radiotracer method for detection of SLN. In 510/529 (96.4%) radiotracer method was used to detect lymph nodes. Blue dye was used in 264/529 (49.9%). Median number of SLN retrieved using both methods were 2. No change in median number of SNL retrieved in different quadrants and central part of the breast were noted, the number remained 2. Conclusions: Our review showed that rate of single lymph node retrieval was lower than what is reported in US SEER data base study of 37%, however the number of median lymph node retrieval was 2 which is considered less than optimal with reported higher rate of false negative rate. We did not attempt to look at disease-specific survival as our sample size is small. Continued data evaluation of established practice is required to improve quality of care and replication of advances established in clinical trials.

Indocyanine Green Fluorescence Versus Blue Dye or Radioisotope Regarding Detection Rate of Sentinel Lymph Node Biopsy and Nodes Removed in Breast Cancer: A Systematic Review and Meta-Analysis.

Background:Either blue dye (BD) or radioisotope (RI) is mainly used for sentinel lymph node biopsy (SLNB) in breast cancer patients. Unlike the BD, RI has lower false-negative rate of SLNB. However, its lymphoscintigraphy, difficulty in preoperative injection, and undetected sentinel lymph nodes in some cases cause surgeons to rely only on BD. Currently, indocyanine green (ICG) fluorescence method (ICG-SLNB) is increasingly used as an alternative to the conventional mapping methods in many centers. This systematic review compared ICG with the conventional method of BD or RI in terms of detection rate of SLNB and the number of sentinel lymph nodes (SLNs) removed in. Methods: We searched all relevant studies published between January 2000 and October 2019. All data on for evaluation of SLN detection rate, number of SLNs removed per patient, and tumor positive rate of SLNB were extracted. Results:A total of 30 studies, including 4,216 SLN procedures were retrieved. There was a statistically significant difference between ICG and BD method in terms of SLN detection rate (OR, 6.73; 95% CI, 4.20-10.78). However, there was no significant difference between ICG and RI in this regard (OR, 0.90; 95% CI, 0.40-2.03). The number of SLNs removed per patient were 2.35 (1.46-5.4), 1.92 (1.0-3.64), and 1.72 (1.35-2.08) for ICG, BD, and RI, respectively. Only in 8 studies, the tumor positive rates in SLNB could be analyzed (ICG, 8.5-20.7%; BD, 12.7-21.4%; RI, 11.3-16%). Conclusion:ICG-SLNB could be an additional or an alternative method for axillary node mapping in breast cancer.

Near Infrared (NIR) Fluorescence is Not a Substitute for Lymphoscintigraphy and Gamma Probe for Melanoma Sentinel Node Detection: Results from a Prospective Trial.

Sentinel lymph node (SLN) biopsy is the standard care for early detection and staging of lymph node metastasis in melanomas. Radiocolloids (RC) and blue dyes are used for SLN detection. Recently, near infrared (NIR) fluorescence tracing using indocyanine green has been developed as an alternative method for SLN detection. The relatively high tissue penetration depth of several millimeters and the ability to detect low concentrations of tracer both suggest that NIR may have significant advantages over RC and the blue dye methods. The objective of this study was to prospectively compare the performance of all three SLN detection techniques using them sequentially to evaluate the same group of patients. One hundred twenty-one primary cutaneous melanoma patients with an indication for SLN biopsy were assigned to the procedure following NIR, blue dye, and RC detection techniques. No adverse event was reported. SLN was not detected in only 4.1% of cases. In 90.9%, an SLN was identified with NIR, but without any auxiliary technique in only 70.2% of cases. RC detected the SLN in 92.6% of cases. Patent blue was found in the sentinel node in 76.9%. The combination of all three techniques detected an SLN in 95.9% of cases. Metastases were present in 26.7%. The false-negative rate was 8.8%, with a negative predictive value of 91.2%. RC was the only technique with high SLN detection. Both the blue dye and NIR methods added sensitivity to the detection rate but should not be a substitute for RC.

PD19-04 VALIDATION OF HYBRID ICG-99mTc-NANOCOLLOID FOR SENTINEL NODE BIOPSY IN A LARGE COHORT OF PENILE CANCER PATIENTS

INTRODUCTION AND OBJECTIVE: Dynamic sentinel node (SN) biopsy is an invasive lymph node (LN) staging method that is typically performed using 99mTc-nanocolloid combined with blue dye to detect micro-metastatic disease for ≥T1G2 penile cancer (PC) patients with no palpable inguinal LNs (cN0). Recent reports suggested that the hybrid tracer indocyanine green (ICG)-99mTc-nanocolloid may improve intraoperative optical SN identification relative to the blue dye. However, this evidence is supported by small sample size series. The aim of the current study was to confirm these findings relying on the largest single center cohort of PC patients who underwent SN biopsy using ICG-99mTc-nanocolloid and blue dye METHODS: 404 patients with ≥T1G2N0 penile squamous cell carcinoma were scheduled for SN biopsy and treatment of the primary tumour at a single European centre. Of those, 255 received SN biopsy using ICG-99mTc-nanocolloid and blue dye. After peritumoural injection of ICG-99mTc-nanocolloid, SNs were preoperatively identified based on lymphoscintigraphy and SPECT/CT. Shortly before surgery, blue dye was administered in all patients using the same injection sites as the hybrid tracer. Intraoperatively, SNs were pursued via gamma tracing, visual identification (blue dye) and near-infrared fluorescence imaging (ICG). Intraoperative SN identification rates of blue dye and ICG-99mTc-nanocolloid were compared using a two-sample t-test RESULTS: There were no tracer-related adverse events. Lymphoscintigraphy and SPECT/CT visualized at least one SN in all patients. All preoperatively defined SNs were intra-operatively identified using a combination of radio-, fluorescence- and blue dye guidance. A total of 841 SNs were excised (median 3 SNs per patient; range 1-9). Of these, 95% were visible with fluorescence imaging, while only 56% were stained by blue dye (p<0.001). Pathologic analyses of the 841 SNs removed, identified 58 positive nodes in 43 patients. Of those, 100% (n=58) and 69% (n=40) were fluorescent or stained blue, respectively. In 45 patients, blue dye did not identify any LN. Of these patients, 7% had positive nodes. Conversely, fluorescence imaging using the hybrid tracer enabled optical detection of at least one SN in all patients CONCLUSIONS: The use of fluorescence improved the optical detection rate of SNs of 39% relative to blue dye. Its use is safe and can replace blue dye for SN biopsy in patients with PC Source of Funding: None

Indocyanine green fluorescence method for sentinel lymph node biopsy in breast cancer

Background/ObjectiveBreast biopsy and analysis of sentinel lymph nodes (SLNs) accurately predict tumor status in the affected basin and help in avoiding unnecessary axillary lymph node dissection, which is associated with remarkable morbidity risk. Blue dye and radioisotope are the most widely used mapping agents, but non-radioactive tracers of comparable accuracy warrant further investigation. This study aimed to investigate utilization of indocyanine green (ICG) fluorescence in sentinel node localization compared with blue dye and to assess the incremental value of ICG. MethodsA total of 39 consecutive patients underwent sentinel lymph node biopsy (SLNB) (40 cases: 38 unilateral and 1 bilateral) with combined blue dye and ICG for localization. The obtained fluorescence images of the lymphatic system were investigated. ResultsAll 84 lymph nodes removed in 40 procedures were identified by ICG, but only 37 were identified by blue dye. The ICG method identified an average of 2.1 SLNs in 39 of 40 cases with a detection rate of 97.5%, but only 0.93 SLN per case with blue dye. Subcutaneous lymphatic channel patterns were also detected by fluorescent imaging in 37 procedures, which all revealed lymphatic drainage toward the axilla except in one case with internal mammary pathway. ConclusionThis study demonstrated the accuracy and safety of ICG for SLNB and its superiority to blue dye method in SLN localization. Therefore, ICG fluorescence method is safe and effective addition in breast clinical settings, wherein blue dye alone is used.

Outcomes of Sentinel Lymph Node Biopsy Using Blue Dye Method for Early Breast Cancer - A Single-Institution Experience in the Philippines.

PurposeThis study aimed to share our experience with SLNB in the Filipino population with early breast cancer.Patients and MethodsA retrospective review was done on all patients with confirmed invasive breast carcinoma, tumor size of 5 cm or less (T1/T2), who preoperatively had no clinical signs of axillary metastasis and subsequently underwent SLNB with blue dye method from January 01, 2008 to December 31, 2017. Clinicopathologic profiles were recorded. Outcomes of patients who had SLNB only were assessed.ResultsOne hundred twenty-nine patients matched the inclusion criteria with a mean age of 54.3 years. The majority (88.4%) had a total mastectomy. Invasive ductal carcinoma (65.1%) was the most common tumor. Estrogen and progesterone receptors were positive in 69% and 61.2% respectively while only 28.7% were HER2 positive. SLNB was successfully carried out in 126 (97.7%) patients with a range of 2–4 SLNs harvested. Thirty-four (26.4%) patients had completion ALND. With a median of 25 months follow-up, 75 out of 95 patients who underwent SLNB alone had follow-up data. Forty-six (61.3%) patients had seroma formation. One (1.3%) patient developed arm paresthesia, 2 (2.7%) local (chest wall) and 2 (2.7%) axillary recurrences after a negative SLNB. None of the patients developed lymphedema.ConclusionThe blue dye method alone is acceptable and can be readily employed in institutions with limited resources. Even with the limited population, the morbidity and oncologic outcomes of patients who underwent SLNB alone were low and comparable to similar international published data. SLNB should be the preferred method for staging the axilla.

Abstract P4-02-07: Validity of omission of axillary lymph node dissection after neoadjuvant chemotherapy for node-positive primary breast cancer

Abstract Background: Anthracycline and taxane-based neoadjuvant chemotherapy (NAC) has been shown to downstage axillary lymph nodes in approximately 40% of patients. However, the feasibility of sentinel lymph node biopsy (SLNB) following NAC for initially node-positive patients is unclear because of high false-negative rates reported in previous trials. The aim of this study was to evaluate whether axillary lymph node dissection (ALND) could be safely omitted for patients with clinically node-positive breast cancer treated with NAC. Patients and methods: We identified 128 patients with clinically node-positive breast cancer who received NAC from March 2006 to March 2017. A correlation between axillary pathologic response and clinicopathological factors was analyzed. Preoperative clinical assessment of lymph-node status was performed by palpation and diagnostic imaging such as ultrasound and MRI. Lymphatic mapping was performed using a combined method of blue dye and radioisotope. Results: The median age was 56.5 (range: 29-79) years and the mean tumor size was 3.6 ± 2.37cm. Of 128 patients, 72 (56.2%) patients had luminal, 32 (31.3%) had HER2-positive, and 24 (18.8%) had triple-negative disease. Sequential anthracycline and taxane were administered for 115 (89.8%) patients, and 25 (78.1%) patients with HER2-positive disease received concomitant trastuzumab preoperatively. Overall, 53 (41.4%) patients achieved axillary pathologic complete response (ypN0). The ypN0 status was significantly correlated with clinical complete response (cCR) in breast (67.9%, p=0.004), ER-negative disease (59.5%, p=0.004) and HER2-positive disease (56.3%, p=0.049). Multivariate analysis demonstrated that cCR in breast (p=0.015) and ER negativity (p=0.006) were significant predictive factors for ypN0. In 14 ER-negative patients achieving cCR in breast, ypN0 was found in 12 (85.7%) patients. Among 85 patients who converted to clinically node-negative, ALND was omitted in 16 patients (18.8%) after SLNB. Of these 16 patients, irradiation to whole-breast and axilla was performed in 14 (87.5%) and 5 (31.3%) patients, respectively. After a median follow-up of 53.2 months, no axillary recurrence was observed in entire patients and 5-year disease-free survival was not significantly different between patients with or without ALND (85.5% vs. 87.5%, p=0.965). Conclusions: As ER-negativity and cCR were correlated with high eradication rate of initially positive lymph nodes, these patients might be good candidates of omission of ALND after NAC. Citation Format: Akiko Matsumoto, Yasuko Umemoto, Daisuke Tsukahara, Hiromitsu Jinno. Validity of omission of axillary lymph node dissection after neoadjuvant chemotherapy for node-positive primary breast cancer [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P4-02-07.

Assessing cytotoxicity and antiproliferation effects of Sida rhombifolia against MCA-B1 and A549 cancer cells

This research aims to determine the cytotoxicity and antiproliferation activities of Sida rhombifolia leaves extract against cancer cells MCA-B1, A549 and normal Vero cells. S. rhombifolia leaves were extracted with ethanol using ultrasonication method and fractionated using n-hexane, ethyl acetate, and water. The tested samples were ethanol extract and n-hexane fraction based on the results of cytotoxicity using the Brine Shrimp Lethality Test (BSLT). The antiproliferation activity test by using Trypan Blue Dye method and the cells harvested after confluence on the third or fourth day and the total cells were calculated by using the Neubauer Hemocytometer. The result showed that the inhibitory activity of ethanol extract at a concentration of 500 ppm is 69.44% with IC50 202.556 ppm on MCA-B1 cancer cells and 69.44% with IC50 276.836 ppm on A549 cancer cells. While the n-hexane fraction at a concentration of 1000 ppm was 64.13% with IC50 425.969 ppm in MCA-B1 cancer cells and 57.18% with IC50 786.617 ppm on A549 cancer cells. After being tested on normal Vero cells the inhibition of normal Vero cells proliferation is not more than 1%. This indicates that ethanol extracts and n-hexane fraction are safe for normal cells and analysis by using LC-MS/MS showed a benzazepine compound in the ethanol extract of S. rhombifolia is known for its role as antiproliferation. These results indicate that S. rhombifolia leaves extract has the potential to be developed as anticancer compounds.

Monte Carlo-based optimization of a gamma probe system for sentinel lymph node mapping

Introduction: Sentinel lymph node biopsy (SLNB) is a standard surgical technique to identify sentinel lymph node (SLN) for the staging of early breast cancer. Nowadays, two methods are used for the identification of SLN: blue dye method aiding visually and radioactive dye using gamma detector. A wide range of gamma probe systems with different design and performance are used in intra-operative surgery. The performance of the probes is evaluated by some parameters such as sensitivity, spatial resolution, angular resolution, and shielding efficiency. Methods: In this study, we simulated a gamma probe system, SURGEOGUIDE II based on CsI(Tl) scintillator, a silicon photomultiplier (SiPM), and a tungsten collimator, using the MCNP4C Monte Carlo (MC) method and comparing with experimental measurement. Finally we modeled a series of probe with various crystal material, crystal length, and collimator hole length to evaluate the sensitivity and the spatial resolution in order to propose the optimal configuration. Results: The sensitivity of the system was measured as 2040 cps/MBq in 30 mm distance from the source. The spatial resolution and angular resolution were 43 mm and 70

Dyeing Method and Properties of a Novel Blue Azo-Anthraquinone Reactive Dye on Cotton.

A novel blue azo-anthraquinone reactive dye was evaluated in the dyeing of cotton by using a dip–pad–steam process. Dyeing method and properties were examined in detail and the results showed that the dyeing method consisting of dye concentration of 25 g/L, sodium carbonate of 12 g/L, dipping time of 3 min and steaming time of 30 min was the most effective when a conventional “one-dip–one-nip” process was used. The fixation of the dyes on cotton could reach up to 93.4%, the wash and rub fastness both reached grade 4 above, and the light fastness reached grade 4–5 above. Such colored cotton showed very close colorimetric properties.

Abstract P3-03-21: Usefulness of sentinel lymph node biopsy by indocyanine green fluorescence method for cN0 breast cancer patients

Abstract Background. Indocyanine green (ICG) fluorescence method (ICG-f) has been recently widely used in sentinel lymph node (SLN) detection. The advantages of ICG-f are no radiation exposure, no limitation to use in high-volume medical centers without radioactive facility, and to confirm lymph flow as a real-time image from outside the body. ICG-f identified an average of 2.3-3.4 SLNs and the detection rate was 99%, compared to 1.7-2 SLNs by RI methods. Long-term observation after SNB using ICG-f has not been reported, including arm lymphedema as the complication of this method.We evaluate the usefulness of SLN biopsy (SNB) for cN0 breast cancer patients from data of multicenter cohort study on long-term results after negative SNB by ICG-f. Methods. Eleven hundred and thirty-two women were enrolled who had histologically proved clinical stage T1-4, pN0, M0 primary invasive breast cancer with SNB using ICG-f (ICG alone or combination of RI/blue dye method) sparing axillary lymph node dissection from May 2007 to December 2015. This study is retrospective, multicenter cohort study conducted at 6 centers in Japan. Primary endpoint is axillary recurrence rate. We analyzed the correlation with the axillary recurrence and adjuvant systemic therapy, adjuvant radiotherapy, and the clinicopathological characteristics. Secondary endpoint is lymphedema. Results and Discussion. The median follow-up time was 41 (range 21-117) months, and axillary recurrence was found in 6 patients (0.53%). Five out of 6 patients were not received standard adjuvant systemic therapy or adjuvant radiation therapy after breast conserving surgerybecause of patient's preference or old age. Lymphedema was identified only 4 patients in 632 patients. It is reported that axillary recurrence after SNB was 0.3-1.65%, which was consistent with our result. Lymphedema was not frequent in patients received SNB using ICG-f, because SLNs are removed along with lymphatic ducts in the limited area of axillary adipose tissue. Conclusion.Axillary recurrence after negative SNB using ICG-f was comparable to RI or blue dye method. It might be important to perform appropriate adjuvant medication or radiation therapy for preventing axillary recurrence after SNB using ICG-f. Next, ICG-f after neoadjuvant chemotherapy is to be investigated, because itis reported that removing more than 2 SLNs were associated with a lower likelihood of false negative ratio in patients with clinically node-positive disease converted to clinically node-negative after chemotherapy, and ICG-f might overcome this issue. Citation Format: Maeshima Y, Takahara S, Yamauchi A, Yamagami K, Sugie T, Yamashiro H, Kato H, Torii M, Takada M, Torii M. Usefulness of sentinel lymph node biopsy by indocyanine green fluorescence method for cN0 breast cancer patients [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P3-03-21.

Abstract P3-03-40: A prospective two arm comparative study of indo-cyanine green (ICG) enhanced fluorescence imaging vs conventional methods (blue dye and radiocolloid/hand held gamma probe) for sentinel lymph node detection in breast cancer - Going beyond the horizon

Abstract Background: The dual technique with radio colloid and blue dye is the gold standard in sentinel lymph node biopsy (SLNB) to stage axilla in breast cancer. However due to cost &amp; infrastructural demands of nuclear medicine department most of the oncology centers are not doing slnb or are doing SLNB with blue dye which is not a standard of care. Indocyanine green (ICG) has recently been used as a method of identifying sentinel lymph nodes. Studies have shown that ICG fluorescence imaging alone or in combination with the blue dye method or the radionuclide method is a safe and easy technique. The objective of the present study was to assess the diagnostic performance of sentinel lymph node (SLN) biopsy using the indocyanine green (ICG) fluorescence method compared with that using the conventional method in detection of sentinel lymph nodes. Material &amp; Methods: 60 patients diagnosed with early breast cancer underwent the SLNB procedure using technetium-99m radio colloid (R), methylene blue dye (MB), and ICG. Fluorescence imaging was done using an indigenously designed , very economical fluorescence imaging system, Irilic.nm fluorescence imaging along with Indocyanine green. All SLNs that were removed during surgery were labelled as hot, blue or/and fluorescent and sent for pathological examination. The detection rate of SLNs and positive SLNs, and the number of SLNs of ICG, MB+ R, ICG + MB, ICG + R were compared. Injection safety of ICG and MB was evaluated. Results: Sentinel Lymph Node was identified in all 60 cases. Total Sentinel lymph nodes removed was 145 (Mean=2, Range 2-5), ICG was able to identify more nodes than the dual dye technique. The identification rate with the dual dye technique was 95%, with blue dye alone 93.6% and with radioisotope alone 96.8% whereas with ICG alone was 100%, with ICG + MB was 96.6% &amp; ICG + R was 96.6 %. 28(46.6%) out of 60 patients had positive nodes which was identified by both dual dye &amp; ICG. None of the patients had any local or systemic reaction with ICG, 3 patients with blue dye had tattooing &amp; staining of skin. Conclusion: ICG is as effective as the dual dye for SLNB. ICG is safe &amp; reliable. In addition, as a near-infrared dye, it has the advantages of real-time visualization, lower cost, and wider availability. It can be a boon for developing countries &amp; second tier referral centers of developed country where there is limited access to nuclear medicine department &amp; radiocolloid and even if its accesible the cost involved is too high which comes with added radiation exposure to medical personnel handling them. A combination of blue dye and ICG is useful dual approach when radioisotope is unavailable. ICG verus Conventional Dye Clinical Profile ICGRadio-colloid+ B;ue DyeDetection Rate100%95%Sensitivity100% (CI 83.16% to 100.00%)100% (CI 83.16% to 100.00%)Positive Predictive Value100%100%Accuracy100%100% Citation Format: Somashekhar S, Rohit Kumar C, Zaveri S, Rajgopal A, Rakshit S, Ali SH. A prospective two arm comparative study of indo-cyanine green (ICG) enhanced fluorescence imaging vs conventional methods (blue dye and radiocolloid/hand held gamma probe) for sentinel lymph node detection in breast cancer - Going beyond the horizon [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P3-03-40.

Abstract P3-03-30: Validity and safety of omission of axillary lymph node dissection among sentinel lymph node-positive breast cancer patients treated with mastectomy

Abstract Background:American College of Surgeons Oncology Group Z0011 trial showed that axillary lymph node dissection (ALND) had no impact on recurrence and survival in patients with positive sentinel lymph node (SLN) after breast-conserving surgery. However, it is still unknown if the omission of ALND can be applicable to patients treated with mastectomy. The aim of this study wasto evaluate whether ALND could be safely omitted for patients with SLN–positive breast cancer after mastectomy. Methods: From a prospective database of 296 patients with clinically node-negative breast cancer who underwent mastectomy and sentinel lymph node biopsy (SLNB) from March 2006 to December 2016, 81 patients who had positive SLNs were selected. Patient characteristics and prognosis were compared between SLN-positive patients with and without ALND. Patients treated with neoadjuvant chemotherapy were excluded from the analysis. Lymphatic mapping was performed using a combined method of blue dye and radioisotope. Results: The median age of entire patients was 57.0 (range: 32-85) years and the median tumor size was 2.5 (range: 0.6-7.9) cm. Of 81 patients, 23 (28.4%) patients omitted ALND. Patients with SLNB alone were more likely to have smaller SLN involvements (p&amp;lt;0.001): micrometastasis was identified in 13 (56.5%) patients in SLNB-alone group and 9 (15.5%) patients in ALND group. The number of positive SLN was comparable between SLNB-alone (median: 1.0, range: 1-6) and ALND groups (median: 1.0, range: 1-5) (p=0.063). There was no significant difference in characteristics including age, tumor size and tumor subtypes between the two groups. Post-mastectomy radiotherapy was performed in 5 (21.7%) patients with SLNB alone and 16 (27.6%) patients with ALND (p=0.588). The majority of patients with macrometastatic SLN received adjuvant chemotherapy in both groups (83.3% vs. 75.5%, p=0.562). Twenty (87.0%) and 51 (87.9%) patients received adjuvant endocrine therapy in SLNB-alone and ALND group, respectively (p=0.584). After a median follow-up of 54.7 months, no axillary recurrence was observed in both groups and 5-year disease-free survival was not significantly different between the two groups (75.0% vs. 88.8%, p=0.489). Lymphedema was observed significantly more often after ALND than after SLNB (22.4% vs. 4.3%, p=0.045). Conclusions: These data suggested that ALND could be safely omitted in SLN-positive breast cancer patients treated with mastectomy and appropriate systemic therapy. Citation Format: Matsumoto A, Umemoto Y, Jinno H. Validity and safety of omission of axillary lymph node dissection among sentinel lymph node-positive breast cancer patients treated with mastectomy [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P3-03-30.