Detection Of Sentinel Lymph Node Metastases Research Articles

Artificial Intelligence-Based Sentinel Lymph Node Metastasis Detection in Cervical Cancer.

Background/objectives: Pathological ultrastaging, an essential part of sentinel lymph node (SLN) mapping, involves serial sectioning and immunohistochemical (IHC) staining in order to reliably detect clinically relevant metastases. However, ultrastaging is labor-intensive, time-consuming, and costly. Deep learning algorithms offer a potential solution by assisting pathologists in efficiently assessing serial sections for metastases, reducing workload and costs while enhancing accuracy. This proof-of-principle study evaluated the effectiveness of a deep learning algorithm for SLN metastasis detection in early-stage cervical cancer. Methods: We retrospectively analyzed whole slide images (WSIs) of hematoxylin and eosin (H&E)-stained SLNs from early-stage cervical cancer patients diagnosed with an SLN metastasis with either H&E or IHC. A CE-IVD certified commercially available deep learning algorithm, initially developed for detection of breast and colon cancer lymph node metastases, was employed off-label to assess its sensitivity in cervical cancer. Results: This study included 21 patients with early-stage cervical cancer, comprising 15 with squamous cell carcinoma, five with adenocarcinoma, and one with clear cell carcinoma. Among these patients, 10 had macrometastases and 11 had micrometastases in at least one SLN. The algorithm was applied to evaluate H&E WSIs of 47 SLN specimens, including 22 that were negative for metastasis, 13 with macrometastases, and 12 with micrometastases in the H&E slides. The algorithm detected all H&E macro- and micrometastases with 100% sensitivity. Conclusions: This proof-of-principle study demonstrated high sensitivity of a deep learning algorithm for detection of clinically relevant SLN metastasis in early-stage cervical cancer, despite being originally developed for adenocarcinomas of the breast and colon. Our findings highlight the potential of leveraging an existing algorithm for use in cervical cancer, warranting further prospective validation in a larger population.

Survival associated with the use of one-step nucleic acid amplification (OSNA) to detect sentinel lymph node metastasis in cervical cancer

IntroductionSentinel lymph node (SLN) biopsy is part of surgical treatment of apparent early-stage cervical cancer. SLN is routinely analyzed by ultrastaging and immunohistochemistry. The aim of this study was to assess the survival of patients undergoing SLN analyzed by one-step nucleic acid amplification (OSNA) compared with ultrastaging. MethodsSingle-center, retrospective, cohort study. Patients undergoing primary surgery and SLN mapping ( ±pelvic lymphadenectomy) for apparent early-stage cervical cancer between May 2017 and January 2021 were included. SLN was analyzed exclusively with OSNA or with ultrastaging. Patients with bilateral SLN mapping failure, with SLN analyzed alternatively/serially with OSNA and ultrastaging, and undergoing neo-adjuvant therapy were excluded. Baseline clinic-pathological differences between the two groups were balanced with propensity-match analysis. ResultsOne-hundred and fifty-seven patients were included, 50 (31.8%) in the OSNA group and 107 (68.2%) in the ultrastaging group. Median follow up time was 41 months (95%CI:37.9–42.2). 5-year DFS in patients undergoing OSNA versus ultrastaging was 87.0% versus 91.0% (p = 0.809) and 5-year overall survival was 97.9% versus 98.6% (p = 0.631), respectively. No difference in the incidence of lymph node recurrence between the two groups was noted (OSNA 20.0% versus ultrastaging 18.2%, p = 0.931). In the group of negative SLN, no 5-year DFS difference was noted between the two groups (p = 0.692). No 5-year DFS and OS difference was noted after propensity-match analysis (87.6% versus 87.0%, p = 0.726 and 97.4% versus 97.9%, p = 0.998, respectively). ConclusionThe use of OSNA as method to exclusively process SLN in cervical cancer was not associated with worse DFS compared to ultrastaging. Incidence of lymph node recurrence in the two groups was not different.

Evaluation of the one-step nucleic acid amplification method for rapid detection of lymph node metastases in endometrial cancer: prospective, multicenter, comparative study

ObjectiveTo evaluate the diagnostic performance of the one-step nucleic acid amplification (OSNA) method for the detection of sentinel lymph node (SLN) metastases in women with apparent early-stage endometrial cancer compared...

Clinical Study of Sentinel Lymph Node Detection to Evaluate Pelvic Lymph Node Metastasis to Determine the Prognosis of Patients with Early Cervical Cancer.

Objective Lymph node status is one of the most important prognostic factors for uterine cervical cancer. Sentinel lymph node (SLN) biopsy has emerged as a potential alternative to systematic lymphadenectomy for the lymph node mapping in such patients. However, the SLN metastasis detection via SLN biopsy in early-stage cervical cancer remains controversial. The current study is aimed at investigating the feasibility and accuracy of combined tracer method for localization of SLN in initial stages of cervical cancer and to evaluate the clinical value of SLN biopsy in replacing pelvic lymph node resection. Methods We retrospectively reviewed 348 cases who were admitted to the Department of Gynecologic Oncology, Shandong Provincial Cancer Hospital, China, between February 2003 and June 2018 with FIGO stage IA2 to IIA2 cervical cancer and undergone through SLN biopsy. Methylthioninium chloride was injected in combination with 99mtechnetium-labeled sulfur colloid prior to surgery to these patients. SLNs were identified intraoperatively, excised, and subsequently submitted to fast frozen section. The detection rates, accuracy, sensitivity, coincidence rate, false negative rate, and negative predictive values of these cases were estimated, and the follow-up outcomes were carefully observed. Chi squared test or Fisher's exact test was employed for a comparison of the categorical variables. Univariate and multivariate Cox proportional hazard models were used for estimation of relationships between overall survival (OS) and disease-free survival (DFS) and prognostic factors. Results The total detection rate of SLN was 97.1% (338/348), and identification of bilateral SLN was successful in 237 patients (70.1%). The patient's tumor size, FIGO stage, lymph node metastasis, and depth of invasion had statistically significant differences in SLN detection rates. The detection rate had inverse relation with tumors size (>4 cm), invasive depth > 2/3, lymph node positive, late staging, and preoperative radiotherapy. 117 positive SLNs were detected in 73 patients. The negative predictive value, sensitivity, false negative rate, and coincidence rate and were 97.7%, 92.4%, 7.6%, and 95.4%, respectively. In patients whose tumor size were ≦ 4 cm, the false negative rate was 4.55% (2/44), whereas it was 0 in patients with tumor size≦2 cm. The respective 1, 3, and 5-year OS was 100%, 94.8%, and 91.8%, respectively, whereas DFS rate for 1, 3, and 5 years was 96.7%, 92%, and 89.6%, respectively. The lymph node was positive, tumor size, the depth of invasion, and staging were statistically different from the recurrence rate and survival rate of patients (p < 0.05). When tumor metastasis exceeded SLN, the recurrence rate was significantly increased, and survival rate is significantly reduced (p < 0.05, p < 0.01, p < 0.05, respectively). Conclusions The identification of SLN combined with 99mtechnetium-labeled sulfur colloid and methylthioninium chloride has a good accuracy and is safe for the assessment of the status of pelvic nodes in patients with initial stage cervical cancer. Nuclide as a tracer has low dependence on objective conditions and doctors' technology and has a good detection rate. In our study, we believe that SLN biopsy is feasible when the tumor is ≦ 4 cm. Large scale clinical trials are required in China expand the sample size and validate the results of this study.

High-resolution three-dimensional imaging for precise staging in melanoma

IntroductionMany cancer guidelines include sentinel lymph node (SLN) staging to identify microscopic metastatic disease. Current SLN analysis of melanoma patients is effective but has the substantial drawback that only a small representative portion of the node is sampled, whereas most of the tissue is discarded. This might explain the high clinical false-negative rate of current SLN diagnosis in melanoma. Furthermore, the quantitative assessment of metastatic load and microanatomical localisation might yield prognosis with higher precision. Thus, methods to analyse entire SLNs with cellular resolution apart from tedious sequential physical sectioning are required. Patients and methodsEleven melanoma patients eligible to undergo SLN biopsy were included in this prospective study. SLNs were fixed, optically cleared, whole-mount stained and imaged using light sheet fluorescence microscopy (LSFM). Subsequently, compatible and unbiased gold standard histopathological assessment allowed regular patient staging. This enabled intrasample comparison of LSFM and histological findings. In addition, the development of an algorithm, RAYhance, enabled easy-to-handle display of LSFM data in a browsable histologic slide-like fashion. ResultsWe comprehensively quantify total tumour volume while simultaneously visualising cellular and anatomical hallmarks of the associated SLN architecture. In a first-in-human study of 21 SLN of melanoma patients, LSFM not only confirmed all metastases identified by routine histopathological assessment but also additionally revealed metastases not detected by routine histology alone. This already led to additional therapeutic options for one patient. ConclusionOur three-dimensional digital pathology approach can increase sensitivity and accuracy of SLN metastasis detection and potentially alleviate the need for conventional histopathological assessment in the future. German clinical trials register(DRKS00015737).

One-Step Nucleic Acid Amplification (OSNA) of Sentinel Lymph Node in Early-Stage Endometrial Cancer: Spanish Multicenter Study (ENDO-OSNA).

Simple SummaryOne-step nucleic acid amplification (OSNA) is an automated molecular diagnostic assay used to detect metastases by analyzing the levels of cytokeratin 19 mRNA in whole lymph nodes. It has been validated as an accurate and reliable tool for staging in several types of cancers and is included in the National Institute for Health and Care Excellence guidelines for the management of breast cancer. ENDO-OSNA is a large, observational, multicenter study designed to evaluate the efficacy of OSNA for the detection of sentinel lymph node (SLN) metastasis in patients with early-stage endometrial cancer. We found that the OSNA assay shows higher sensitivity, specificity, and diagnostic accuracy in the detection of SLN metastasis, including low-volume metastasis, compared to standard pathological ultrastaging. Moreover, OSNA could aid in the identification of patients with intermediate or high-risk endometrial cancer, and lead to treatment decisions that could improve their prognosis.The objective of this study was to evaluate the efficacy of one-step nucleic acid amplification (OSNA) for the detection of sentinel lymph node (SLN) metastasis compared to standard pathological ultrastaging in patients with early-stage endometrial cancer (EC). A total of 526 SLNs from 191 patients with EC were included in the study, and 379 SLNs (147 patients) were evaluated by both methods, OSNA and standard pathological ultrastaging. The central 1 mm portion of each lymph node was subjected to semi-serial sectioning at 200 μm intervals and examined by hematoxylin–eosin and immunohistochemistry with CK19; the remaining tissue was analyzed by OSNA for CK19 mRNA. The OSNA assay detected metastases in 19.7% of patients (14.9% micrometastasis and 4.8% macrometastasis), whereas pathological ultrastaging detected metastasis in 8.8% of patients (3.4% micrometastasis and 5.4% macrometastasis). Using the established cut-off value for detecting SLN metastasis by OSNA in EC (250 copies/μL), the sensitivity of the OSNA assay was 92%, specificity was 82%, diagnostic accuracy was 83%, and the negative predictive value was 99%. Discordant results between both methods were recorded in 20 patients (13.6%). OSNA resulted in an upstaging in 12 patients (8.2%). OSNA could aid in the identification of patients requiring adjuvant treatment at the time of diagnosis.

Abstract PS1-04: Utility of rapid- immunohistochemistry using an alternating current electric field for intraoperative diagnosis of sentinel lymph nodes in breast cancer

Abstract Axillary lymph node status and pathological diagnosis of sentinel lymph nodes (SLNs) is an important factor that influences management of postoperative therapy. Recent reports indicate that one-step nucleic acid amplification (OSNA) and hematoxylin and eosin (HE)-stained frozen sections are effective for intraoperative diagnosis of SLNs. In the present study, we report a rapid-immunohistochemical staining (R-IHC) method that enables intraoperative detection of SLN metastases within 16 min using an anti-cytokeratin antibody. With this R-IHC system, we apply a high-voltage, low-frequency AC electric field to lymph node sections while they are incubating with the antibodies. The antibodies are mixed within microdroplets and the opportunity for contact between the antibody and antigen is increased. This greatly reduces the time required for the antigen-antibody reaction. This is the report on SLN diagnosis using R-IHC in patients with breast cancer. We prospectively examined 632 dissected SLNs from 260 breast cancer patients who underwent surgery at our institute between July 2014 and March 2020. The dissected SLNs were sectioned and conventionally stained with HE or immunohistochemically labeled with anti-cytokeratin antibody using R-IHC procedures. Intraoperative R-IHC analyses were completed within 16 min, after which diagnoses were made by two pathologists. The total time required for intraoperative diagnosis was about 20 min. In this study, R-IHC detected four metastatic SLNs that were undetected using conventional HE staining (10/52, 19.2%). Compared with subsequent permanent diagnosis, R-IHC offered 98.0% sensitivity, 100% specificity and 99.8% accuracy while those values for intraoperative HE were 80.4%, 100%, 98.4%, respectively (isolated tumor cells were counted as negative). These findings indicate R-IHC is a clinically applicable technique that enables precise, and quick intraoperative detection of micro- and macrometastases in breast cancer. Moreover, R-IHC is cost-effective method: it’s cost is less than a quarter of OSNA and previous report showed that the concentration of primary antibody could be reduced by more than 90% by using R-IHC because AC mixing activated antigen-antibody reaction. Another benefit is its ability to shorten the time needed for microscopic diagnosis itself. When using R-IHC, pathologists are able to readily find metastatic lesions, even within low-power fields (e.g. 40x). This reduces not only the time needed for intraoperative diagnosis, but also the effort necessary to find unclear lesions like small cancer cells (even though it is macrometastasis), micrometastasis, and artifacts of frozen section. This study was conducted at a single institute. Further investigation in a multi-institutional collaborative prospective study will be needed to confirm the utility of this method. But for now, R-IHC appears to be a cost-effective and clinically applicable method for diagnosis of breast cancer and SLN metastasis. Citation Format: Kaori Terata, Hiroshi Nanjo, Eriko Takahashi, Ryuta Nakamura, Yoichi Akagami, Ayuko Yamaguchi, Misako Yatsuyanagi, Chiaki Kudo, Yoshihiro Minamiya. Utility of rapid- immunohistochemistry using an alternating current electric field for intraoperative diagnosis of sentinel lymph nodes in breast cancer [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS1-04.

Diagnostic criteria of sentinel lymph node micrometastasis or macrometastasis based on tissue rinse liquid-based cytology in gynecological cancer.

This study aimed to establish intraoperative diagnostic criteria of sentinel lymph node (SLN) micro-/macrometastasis on the basis of tissue rinse liquid-based cytology (TRLBC) in gynecological cancer. We enrolled 214 patients with gynecological cancer who underwent rapid diagnosis of SLN metastasis on the basis of TRLBC from a total of 490 SLNs. For slides that were classified as positive for atypical cells on cytological inspection, we counted the number of clusters (an atypical cell mass consisted of three or more cells) and the number of single cells (an atypical cell other than clusters). Receiver operating characteristic (ROC) analysis was applied to determine the efficiency of predicting SLN micro-/macrometastasis. On cytological inspection, 36 slides were classified as positive for atypical cells, while 21 slides (4.3%) were true positive, 15 (3.1%) were false positive, and 454 (92.6%) were true negative. There were no false negative results in this study. The area under the ROC curve for the number of cluster was superior to that for the number of single cells for distinguishing micro-/macrometastasis from negative/isolated tumor cells (0.86 vs. 0.67, P = 0.032). The optimum cut-off value of the number of clusters was 5 for distinguishing these two categories. TRLBC is a highly sensitive alternative for detecting SLN metastasis as a rapid intraoperative diagnosis. Counting the number of atypical cell clusters might be useful for distinguishing micro-/macrometastasis from isolated tumor cells.

Intraoperative imprint cytology of sentinel lymph nodes in breast cancer patients: Comparation with frozen section

Background/Aim. Sentinel lymph node (SLN) biopsy has been established as the standard of care for axillary staging in patients with invasive breast carcinoma and clinically negative lymph nodes. Intraoperative assessment of sentinel lymph nodes might be done by frozen section (FS), touch imprint cytology (TIC) and one step nucleic acid amplification. The aim of this study was to review our institution's results with SLN biopsy using TIC and FS technique as intraoperative diagnostic tool for breast cancer patients. Methods. SLNs from 101 patients were examined intraoperatively by frozen hematoxylin-eosin (H&amp;E) stain and by touch imprint cytology. Results of TIC were compared with FS and permanent histology sections. Results. The total number of dissected SLNs was 163 with a mean of 1.6 (1?4) per patient. The permanent H&amp;E identified 19 (19%) patients with a sentinel lymph node metastasis and 82 (81%) patients with tumor-free sentinel nodes. The sensitivity/ specificity rates were 73.7%/99.3%, respectively for TIC and 84.2%/100%, respectively for FS. Relevant positive/ negative predictive values were 93.3%/96.6%, respectively for TIC and 100%/97.9%, respectively for FS. Conclusion. Our experience with TIC and FS for the intraoperative evaluation of SLNs is similar to the findings from previously reported studies. We detected the high specificity for both methods, but TIC technique appeared to be less sensitive than FS in detecting SLN metastases in breast cancer patients. TIC could be recommended as reasonable alternative to frozen section due to its simplicity and low cost.

Accuracy of One-Step Nucleic Acid Amplification in Detecting Lymph Node Metastases in Endometrial Cancer.

One-step nucleic acid amplification (OSNA) is used to intraoperatively detect sentinel lymph node metastases in breast cancer. OSNA has also been proposed in endometrial cancer, but evidence in this regard is unclear to define the diagnostic accuracy of OSNA in detecting lymph node metastases in endometrial cancer. A systematic review and meta-analysis was performed by searching 8 electronic databases from their inception to March 2019 for studies testing the diagnostic accuracy of OSNA in detecting sentinel lymph node metastasis in endometrial cancer. Pathologic ultrastaging was the reference standard. Sensitivity, specificity, positive and negative likelihood ratio (LR+ and LR-), diagnostic odds ratio (DOR) and area under the curve (AUC) on SROC curve were calculated. Four studies with 237 patients and 691 lymph nodes were included. OSNA showed sensitivity = 0.88, specificity = 0.93, LR + =17.95, LR- = 0.15, DOR = 191.23 and high diagnostic accuracy (AUC = 0.959). OSNA appears as a highly accurate tool for intraoperative assessment of sentinel lymph node in endometrial cancer.

Abstract P3-03-29: Evaluation of a direct reverse transcription loop-mediated isothermal amplification method without RNA extraction (direct RT-LAMP) for the detection of lymph node metastasis in early breast cancer

Abstract Backgroud: The detection of lymph node metastasis by reverse transcription loop-mediated isothermal amplification method (RT-LAMP) had been studied previously. Even though, RT-LAMP method provides improved performance compared to intraoperative histology sentinel lymph node (SLN) evaluation, direct RT-LAMP method without RNA extraction can be more efficient and easily accessible process. Therefore, we evaluated the performance and efficacy of a direct reverse transcription loop-mediated isothermal amplification (direct RT-LAMP) assay for visual detection of CK19, CK20, and CEA mRNAs to identify lymph node metastasis in patients with early breast cancer. Methods: A total of 92 lymph nodes dissected from 40 patients with breast cancer were collected at the breast cancer center of Kyungpook National University Chilgok Hospital between November 2015 and February 2016. All of the samples were analyzed by direct RT-LAMP assay and routine histopathology examination. Cutoff values to distinguish metastasis and nonmetasis were determined by measuring cytokerain 19 (CK19) mRNA in histopathologically positive and negative lymph node using direct RT-LAMP. Results: We set the cutoff value of direct RT-LAMP assay for CK 19 mRNA at 1ng to distinguish status of LN metastasis. The sensitivity and specificity of the RT-LAMP assay were 85.7% and 100%, respectively. The positive predictive value and negative predictive value were 100% and 94.4%. Conclusion: Direct RT-LAMP assay can allow detection of SLN metastasis in breast cancer patients intraoperatively with a good sensitivity through cost-effective and time–saving manner. Citation Format: Lee IH, Jung J, Lee S, Lee J, Lee RK, Park H, Jung J, Kang J, Chae Y. Evaluation of a direct reverse transcription loop-mediated isothermal amplification method without RNA extraction (direct RT-LAMP) for the detection of lymph node metastasis in early breast cancer [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 P3-03-29.

Abstract P3-03-19: Tumor cell detection and immune profiling of lymph nodes from breast cancer patients by mass cytometry

Abstract Introduction: A sentinel lymph node (SN) is the primary node draining the tumor and is assumed to be affected early in the metastatic process. Detection of metastases in SN is a standard procedure in breast cancer diagnostics based on microscopic evaluation (morphology and immunohistochemistry), determining the need for removal of all axillary glands for inspection which again is crucial for tailoring adjuvant therapy. The identification by microscopy is time-consuming and has a risk for false negative results. We hypothesize that the immune profile of SN changes with the presence of tumor cells, even at very low frequencies (micrometastases). By using a multi marker approach to characterize millions of cells from sentinel lymph nodes with and without metastases we aimed at identifying both tumor cells but also characterize a tumor specific immune response. This dual approach might provide an opportunity for a more sensitive test for SN diagnostics. Material and Methods: We established a mass cytometry assay containing 38 markers (antibodies) using CyTOF technology to combine immune profiling with identification of breast cancer cells. Cell suspensions from 14 metastatic axillary lymph nodes (ALNmet), 16 metastatic sentinel lymph nodes (Snmet) and 14 non-metastatic sentinel lymph nodes (SN) from breast cancer patients from the clinical observational trial Oslo2 (early, operable breast cancer patients representing all subtypes) were successfully analyzed by the multimarker panel (single cell resolution). Results: By using mass cytometry, we detected tumor cells (gated as PanKeratin+/CD45- cells) in 86% (26/30) metastatic lymph nodes (ALNmet and Snmet) and in 14% (2/14) non-metastatic lymph nodes (SN). Further, the leukocyte population, identified as CD45+ cells, was gated into 15 subpopulations, mainly comprising different subsets of B and T cells, monocytes and NK cells. By comparing the leukocyte composition in the ALNmet with those in SN samples we identified a significant increase in the abundance of CD8+ memory phenotype, TFH and TCRγδ cells and a decrease in the CD4+ subpopulation in ALNmet compared to the SN samples. The Snmet samples had smaller deposits of tumor cells than the ALNmet samples, and we found no significant differences in leukocyte composition between Snmet and SN samples. Interestingly, when looking at the activation marker CD56, we observed a significant higher expression in the CD4RO, CD8RO, TFH and Treg subpopulations of Snmet samples compared to SN samples. Conclusion: In this study we identified a significant difference in immune cell composition in lymph nodes with and without metastases (ALNmet compared to SN samples). We also identified activation markers unique for subpopulations of lymphocytes in Snmet, but not in negative lymph nodes (SN). We were also able to detect and identify micrometastases in most lymph nodes where morphological examination had identified them, but in addition found tumor cells in two samples scored as negative. The results will be validated in a larger sample series. Citation Format: Russnes HG, Rye IH, Huse K, Schlichting E, Garred O, Mykelbust JH. Tumor cell detection and immune profiling of lymph nodes from breast cancer patients by mass cytometry [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 P3-03-19.

One-step nucleic acid amplification assay is an accurate technique for sentinel lymph node biopsy of breast cancer patients: a meta-analysis

Background:To estimate the accuracy of one-step nucleic acid amplification (OSNA) assay as an intra-operative sentinel lymph node biopsy (SLNB) for sentinel lymph node (SLN) metastasis in breast cancer.Methods:PubMed, Cochrane Library and Web of Science databases were searched by two independent reviewers to retrieve literature with per-patient analysis. The deadline was up until December 2016. A meta-analysis was performed using STATA, Meta-Disc, and Revman software. A random-effects model was used and subgroup analysis was carried out to identify possible sources of heterogeneity.Results:According to the inclusion criteria, 2833 patients from 12 studies were included in this meta-analysis. The pooled sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR) and the area under the sROC curve (AUC) for detecting SLN metastasis were 0.87 (95% CI 0.81–0.91), 0.92 (95% CI 0.86–0.95), 10.65 (95% CI 6.18–18.34), 0.14 (95% CI 0.10–0.20), 75.08 (95% CI 37.77–149.22) and 0.94 (95% CI 0.91–0.95), respectively.Conclusions:The present study adds the evidence that OSNA assay is an accurate molecular diagnostic tool for intra-operatively detecting SLN metastasis in breast cancer. One-step nucleic acid amplification assay might be introduced into clinical usage for replacing traditional intro-operative diagnostic methods of SLNB.

Exhaustive pathologic work-up in sentinel lymph node biopsy for melanoma: is it necessary?

The objective of this study is to determine whether a less exhaustive pathologic work-up to detect melanoma metastasis is clinically useful and does not affect patient prognosis. The success and evolution of the sentinel lymph node (SLN) depends on histological techniques. Several exhaustive protocols of SLN analysis have been published, but are time and cost consuming, with slight increases in the rates of metastasis detection. From 281 patients with SLN biopsy, each SLN was sectioned every 2 mm and from each paraffin block, 2-3 histological sections were evaluated. The patients were divided as follows: the first group (n=185) was subjected to extensive SLN examination (eSLNe) and the second group (n=96) was not subjected to an extensive SLN examination (wSLNe). The average SLN resected was 2 (range: 1-7), evaluating one in 50.9%. The SLN metastasis detection rate was 28.5%, whereas eSLNe increased by 3.2%. During follow-up, 4/26 (17.4%) cases in the wSLNe group showed recurrence in the SLN basin. Factors associated with decreased survival in univariant analysis were recurrence, Breslow thickness, advanced clinical stage, ulceration, and SLN metastasis. eSLNe did not affect disease-specific survival. Multivariate analysis showed recurrence (hazard ratio 23.475, 95% confidence interval: 1903-4559, P<0.001) and Breslow thickness of more than 3.5 mm (hazard ratio 15.222, 95% confidence interval: 1448-3059, P<0.001) as independent risk factors for decreased survival. Our routine for SLN examination enabled an adequate rate of SLN metastasis detection and the eSLNe increased the rate of detection in 3.2%, but did not affect the survival. We did not find any benefit from performing the eSLNe in patients with Breslow thickness less than 3.5 mm.

ASSESSMENT OF SENTINEL LYMPH NODE METASTASIS IN CANINE MAMMARY GLAND TUMORS USING COMPUTED TOMOGRAPHIC INDIRECT LYMPHOGRAPHY.

In the staging process of the breast cancer, demonstrating metastasis of the sentinel lymph node (SLN) has an important prognostic value, in both humans and animals. The aim of this prospective case-control study was to determine the diagnostic value of computed tomographic indirect lymphography (CT-LG) for detecting SLN metastasis in dogs with mammary cancer. Thirty-three female dogs with tumors in the abdominal and inguinal mammary glands were prospectively selected and subjected to CT-LG, 1 and 5 min after injection of 1 ml of contrast agent (iopamidol) in the subareolar tissue of the neoplastic and the contralateral normal mammary glands. The pattern of postcontrast opacification, degree of postcontrast enhancement, and size and shape were assessed in 65 SLNs in total and were correlated with histopathological findings. The absence of opacification or heterogeneous opacification 1 min after contrast medium injection showed the highest sensitivity, specificity, and accuracy (93%, 100%, and 98.4%, respectively). In images taken 1 min after injection, an absolute density value lower than 444 Hounsfield units (HU) in the center of the SLN also provided significant sensitivity and specificity (93.8% and 75%, respectively). The size and shape of the SLN (maximum and minimum diameter, maximum/minimum diameter ratio, maximum diameter/height of fifth thoracic vertebral body ratio) showed the lowest sensitivity and specificity. Results of this study support the hypothesis that CT-LG could help in the assessment of SLN metastasis in cases of mammary gland tumors in dogs.

One step nucleic acid amplification (OSNA) study in Indonesia

ABSTRACT Sentinel lymph node (SLN) is defined as the first of a few selected lymphatic nodes, into which lymphatic fluid from a primary tumor drains. Streamlined processing of sentinel lymph nodes (SLN) for detection of lymph node metastasis involves the able command over methodical blocks of SLN identification, surgical removal of SLN and SLN analysis. One Step Nucleic Acid Amplification (OSNA) method, which relies on CK19 mRNA expression to detect intraoperatively lymph node metastases in breast cancer cases, emerged as a plausible alternative to the current gold standard that uses histopathological node analysis. Sixty selected axillary sentinel lymph nodes from thirty breast cancer patients. Sentinel lymph nodes were directly bi-halved after collection using customized lymph node cutting device (Sysmex), or scalpel. The first halves were subjected to histopathological examination and were stored in specimen containers containing fresh formaldehyde prior to processing. The adjacent halves were weighed to comply with the required mass by OSNA detection in the range of 50 – 600 mg and wrapped in clean foils for storage in -80°C prior to OSNA analysis. 60 SLNs were same diagnosis using both methods. 25 SLNs were negative and 25 SLNs were positive using both methods. 3 SLNs were positive on OSNA but negative on histology. Other 7 SLNs were negative on OSNA but positive on histology, and these 1 nodes contained only micrometastasis lesion. These results suggest that OSNA is a useful for detecting SLNs metastasis, but a copy number of CK19 might be an indepedent factor from prediction and prognosis of breast cancer.

The Prognostic Significance of Sentinel Lymph Node Status for Patients with Thick Melanoma.

Sentinel lymph node biopsy (SLNB) is recommended for patients with intermediate-thickness melanoma, but the use of SLNB for patients with thick melanoma is debated. This report presents a single-institution study investigating factors predictive of sentinel lymph node (SLN) metastasis and outcome for thick-melanoma patients . A retrospective review of a single-institution database from 1997 to 2012 identified 147 patients with thick primary cutaneous melanoma (≥4mm) who had an SLNB. Clinicopathologic characteristics were correlated with nodal status and outcome. The median age of the patients was 67years, and 61.9% of the patients were men. The median tumor thickness was 5.5mm, and 54 patients (36.7%) had a positive SLN. Multivariable analysis showed that only tumor thickness significantly predicted SLN metastasis (odds ratio 1.14; 95% confidence interval (CI) 1.02-1.28; P=0.02). The overall median follow-up period was 34.6months. Overall survival (OS) and melanoma-specific survival (MSS) were significantly worse for the positive versus negative-SLN patients. Multivariable analysis showed that age [hazard ratio (HR) 1.04; 95% CI 1.01-1.07; P=0.02] and SLN status (HR 2.24; 95% CI 1.03-4.88; P=0.04) significantly predicted OS, whereas only SLN status (HR 3.85; 95% CI 2.13-6.97; P<0.01) significantly predicted MSS. Tumor thickness predicts SLN status in thick melanomas. Furthermore, SLN status is prognostic for OS and MSS in thick-melanoma patients, with positive-SLN patients having significantly worse OS and MSS. These findings show that SLNB should be recommended for thick-melanoma patients, particularly because detection of SLN metastasis can identify patients for potential systemic therapy and treatment of nodal disease at a microscopic stage.

One-step nucleic acid amplification (OSNA) for the detection of sentinel lymph node metastasis in endometrial cancer

ObjectiveTo evaluate the efficacy of one-step nucleic acid amplification (OSNA) for the diagnosis of sentinel lymph node (SLN) metastasis compared with histopathological examination in patients with endometrial carcinoma. MethodsA total of 94 SLNs from 34 patients with endometrial carcinoma were enrolled. The central 1-mm portion of each node was subjected to semi-serial sectioning, sliced at 200-μm intervals and examined by hematoxylin and eosin and cytokeratin 19 (CK19) immunohistochemical staining, and the remaining tissue was analysed by OSNA using CK19 mRNA. The accuracy of the OSNA assay was evaluated based on histopathological diagnosis. ResultsHistologically, 89 SLNs were determined to be metastasis negative, and the remaining five SLNs were metastasis positive. Using the breast cancer cutoff value for detecting lymph node metastasis (OSNA criteria for breast cancer, >250copies/μl) the sensitivity of the OSNA assay was 100%; specificity was 87.6%; diagnostic accuracy was 88.3%. Discordant results were recorded for 11 of 94 SLNs. In all 11 cases, a positive result was given by the OSNA assay but not by histopathological examination. In two SLNs from the same patient, histopathological examination revealed the presence of benign epithelial inclusions that were CK19 positive; both SLNs yielded a positive result in the OSNA assay (true–false positive). All remaining nine histologically-negative/OSNA-positive SLNs were classified as micrometastasis (+) by the OSNA assay. ConclusionThe OSNA assay shows high sensitivity and specificity, which suggests its utility as a novel tool for the molecular detection of SLN metastasis in patients with endometrial carcinoma.

Intraoperative detection of sentinel lymph node metastases inbreast carcinoma by Fourier transform infrared spectroscopy.

Sentinel lymph node (SLN) biopsy is a routine surgical staging procedure in clinically lymph node-negative breast cancer. Fourier transform infrared (FTIR) spectroscopy, a technique based on the biochemical composition of the tissue, has previously been found to be capable of differentiating between normal and malignant tissue. The aim of the present study was to explore the intraoperative use of FTIR spectroscopy for rapidly identifying metastatic SLNs, and distinguishing between metastatic and non-metastatic tissue. Freshly removed SLNs from patients with breast cancer were analysed. Samples were measured by FTIR spectroscopy before histopathological diagnosis. The FTIR spectrum of each sample identified ten bands from 2000 to 900 cm(-1) . The peak position, intensity and full width at half maximum of each absorbent band were measured, and the relative intensity ratios calculated. Canonical discriminant analysis was performed to discriminate between metastatic and non-metastatic samples. A total of 149 SLNs were removed from 49 patients. Histopathological examination confirmed 38 metastatic and 111 non-metastatic SLNs. Eighteen of 29 parameters were significantly different between the metastatic and non-metastatic SLNs. Five parameters were selected as independent factors to form discriminant functions. The sensitivity, specificity and accuracy of this method were 94·7, 90·1 and 91·3 per cent respectively. The accuracy of histological analysis of frozen sections was 100 per cent. FTIR spectroscopy is a promising technique for the real-time diagnosis of SLN metastasis during breast cancer surgery. Surgical relevance Sentinel lymph node (SLN) biopsy is a highly accurate predictor of overall axillary status and has become the standard in disease staging in clinically node-negative breast cancer. A rapid and accurate intraoperative assessment of metastatic spread to the SLN provides the necessary information for the surgeon to proceed with immediate axillary dissection. The results of this research indicate that Fourier transform infrared (FTIR) spectroscopy is a rapid, accurate, non-destructive and cost-effective molecular method that can be used to detect SLN metastasis during surgery. FTIR analysis could be useful for the intraoperative diagnosis of lymph node metastases at large institutions, thereby reducing the workload of pathologists, as well as in regions lacking pathologists such as in developing countries.

Efficacy and feasibility of the immunomagnetic separation based diagnosis for detecting sentinel lymph node metastasis from breast cancer.

A purpose of this study was to establish a novel molecular diagnostic model and provide new insight into the intraoperative evaluation of the sentinel lymph node (SLN) metastasis in breast cancer. A total of 124 breast cancer patients who met the criteria of sentinel lymph node biopsy (SLNB) and underwent intraoperative biopsy were consecutively enrolled in this study. After the SLNs obtained from each patient were labeled, MOC-31 monoclonal antibody-mediated immunomagnetic separation (IMS) and flow cytometry were used to determine the expressions of breast cancer metastasis-related markers, including Mucin 1 (MUC1), CD44v6, and HER2. Alternatively, conventional intraoperative hematoxylin and eosin (HE) staining and cytokeratin immunohistochemistry (CK-IHC) were performed to detect potential SLN metastasis. The sensitivity, specificity, and false-negative rate of the three intraoperative diagnostic methods were compared and analyzed. A total of 55 positive-SLNs were found in 38 breast cancer patients using IMS, yielding a sensitivity of 86.4% (38/44), specificity of 94.7% (36/38), accuracy of 93.5% (116/124), false-positive rate of 2.5% (2/80), false-negative rate of 13.6% (6/44), positive predictive value of 95.5% (42/44), and negative predictive value of 93.0% (80/86). Patients with high expressions of CD44v6, MUC1, and HER2 in SLNs tended to have higher number of positive lymph nodes, among which the MUC1 and HER2 showed significant differences (P<0.05). Therefore, compared with conventional HE staining and CK-IHC, IMS technology has remarkably higher sensitivity and specificity and relative lower false-negative rate, thus making it an effective and feasible intraoperative detection method of SLN for breast cancer diagnosis to some extent.