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We evaluated the prognostic implications of the circulating tumor cell (CTC) count in non-metastatic, HER2-negative breast cancer patients who failed to achieve pathologic complete response (pCR) after neoadjuvant chemotherapy (NCT). A total of 173, non-metastatic breast cancer patients treated with NCT were prospectively enrolled. CTCs were obtained from blood drawn pre-NCT and post-NCT using a SMART BIOPSY SYSTEM isolation kit (Cytogen Inc., Seoul, Korea) with immunofluorescence staining. Excluding 26 HER2-positive patients, Relapse-free survival (RFS) and overall survival (OS) related to the CTC count and the association of the CTC count with the treatment response to given therapy were analyzed in 147 HER2-negative patients. Among 147 HER2-negative patients, 28 relapses (19.0%) and 13 deaths (8.8%, all breast cancer-specific) were observed during a median follow-up of 37.3 months. One hundred and seven patients (72.8%) were hormone receptor-positive, and 40 patients (27.2%) had triple-negative breast cancer (TNBC). One or more CTCs were identified in 88 of the 147 patients (59.9%) before NCT and 77 of the 134 patients (52.4%) after NCT. In the entire HER2-negative patient cohort, the initial nodal status was the most significant factor influencing RFS and OS. In TNBC, 11 patients (27.5%) achieved pCR and patients that failed to achieve pCR with ≥ 5 CTCs after NCT, showed worse RFS (HR, 10.66; 95% CI, 1.80–63.07; p  = 0.009) and OS (HR, 14.00; 95% CI, 1.26–155.53; p  = 0.032). The patients with residual tumor and a high number of the CTCs after NCT displayed the worse outcome. These findings could provide justification to launch a future, well designed trial with longer follow-up data to obtain regulatory approval for clinical use of the assay, especially for the ER-positive, HER2-negative breast cancer subset.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Photosynthetic organisms shift the dynamics of surface pCO 2 driven by the sea surface temperature change (thermodynamic driver) by assimilating inorganic C from seawater. Here we measured net C uptake in a macroalgal habitat of coastal Korea for two years (2019-2020) and found that the macroalgal habitat contributed 5.8 g C m −2 month −1 of the net C uptake during the growing period (the cooling period, September−May). This massive C uptake changed the thermodynamics-driven seasonal dynamics such that the air−sea equilibrium of pCO 2 was pushed into disequilibrium. The surface pCO 2 dynamics during the cooling period was mostly influenced by the seasonal decrease in temperature and the proliferation of macroalgae, while the dynamics during the warming period (the stagnant period, June−August) closely followed that predicted based solely on the change in sea surface temperature (thermodynamic driver). In contrast to the phytoplankton-dominated off-shore waters (where phytoplankton populations are large in spring and summer), the impact of coastal macroalgae on surface pCO 2 dynamics was most pronounced during the cooling period, when the magnitude of pCO 2 change was as much as twice that resulting from temperature change. Our study shows that the distinctive features of the macroalgal habitat—in particular the seasonal temperature extremes (~18°C difference), the active macroalgal metabolism, and anthropogenic nutrient inputs—collectively influenced the seasonal decoupling of seawater and air pCO 2 dynamics.

Urchin barrens have been a major issue of rocky coastal ecosystems in temperate regions. In South Korea, the east coast and Jeju Island have especially been a focus because the area of barren ground increases in spite of continual efforts to install artificial reefs. This study approached the urchin barrens issue in South Korea, by focusing on a correlational analysis of urchin and macroalgal abundance. Urchin density and algal species coverage were obtained using a quadrat image analysis. Subtidal sites were then classified into three groups according to the average densities of urchins to evaluate the characterization of the macroalgal community: no urchin zone; transition (TR) zone, 4 inds. m-^sup 2^; and urchin zone, ≥8 inds. m-2. The significant correlations indicate that the abundance of some macroalgal species is proportionally regulated by sea urchin density. This study also shows how macroalgal vegetation changes in response to an urchin's density gradient in a natural condition; and there is a TR zone that existed with respect to an intermediate level of algal abundance.

Background/Aim: Although it has been suggested that circulating tumor cells (CTCs) and circulating tumor DNA (ctDNA) might be used in a complementary manner in lung cancer diagnosis, limited confirmatory data are available. In this prospective study, we evaluated the diagnostic performance of each assay separately and in combination. Patients and Methods: From March 2018 to January 2019, patients with suspected primary lung cancer, who underwent routine lung cancer work-up and peripheral blood sampling, were prospectively enrolled in the study. Epithelial cell adhesion molecule and cytokeratin served as markers of CTCs. In terms of ctDNA analysis, single-nucleotide variants were evaluated via next-generation sequencing. Results: We analyzed 111 patients, including 99 with primary lung cancer and 12 with benign pulmonary disease. The median number of CTCs in 10 ml of blood was 3. The most frequently detected single nucleotide variants of ctDNA were TP53, CDKN2A, and EGFR. The diagnostic sensitivity of conventional tumor marker (combination of carcinoembryonic antigen/CYFRA 21-1/neuron-specific enolase) was 66.7%, while those of the ctDNA and CTC assays were 72.7% and 65.7%, respectively. The sensitivity of the CTC/ctDNA combination (95.0%) was significantly greater than those of the CTC (p<0.001), ctDNA (p<0.001), or conventional tumor marker (p<0.001) alone. Subgroup analysis revealed that the sensitivity of the combination assay was greater than those of the CTC or ctDNA assays alone, regardless of tumor stage or histopathology type. Conclusion: The CTC/ctDNA combination assay enhanced the sensitivity of primary lung cancer diagnosis. The combination assay strategy may be clinically useful and could enhance the early detection of lung cancer (ClinicalTrials.gov number: NCT03479099).

The sea urchin, Mesocentrotus nudus, is widely distributed in North West Pacific regions. It has a substantial impact on macroalgal communities as a generalist herbivore. This study examined various aspects of its feeding ecology, including algal preference, foraging behaviors, and possible effects of past feeding history on its algal preference. We used six common algal species (Ulva australis, Undaria pinnatifida, Sargassum confusum, Dictyopteris divaricata, Grateloupia elliptica, and Grateloupia angusta) from the east coast of Korea as food choice in a series of indoor aquarium experiments. The first choice of starved M. nudus was exclusively U. pinnatifida, followed by G. elliptica and S. confusum. Unlike large urchins, small urchins equally preferred U. pinnatifida and G. elliptica. On the other hand, Undaria-fed urchins preferred to feed only G. elliptica, although its preference slightly differed over time. We then grouped sea urchins into three categories (starved, Undaria-fed, mixed species-fed) to observe 12-days feeding preference as well as early foraging movements. Foraging behaviors of the three groups were distinctively different, although they could not completely reflect the actual consumption. For example, U. australis was highly attractive, but rarely eaten. Undaria-fed urchins seemed to stay with only S. confusum and U. australis. This study demonstrates that M. nudus shows high flexibility in food preference depending on past feeding history and body size. Its foraging behaviors are also affected by past feeding conditions, exhibiting active chemoreceptive movements.

To evaluate the clinical efficacy of a circulating tumor cell (CTC) test by comparison between healthy volunteers and patients with localized prostate cancer including those under active surveillance. CTC counts in peripheral blood were compared between patients with prostate cancer (n=45) and healthy volunteers (n=17). CTCs were identified based on the expression of epithelial cell adhesion molecule (EpCAM) and counted using a SMART BIOPSY™ SYSTEM. The number of EpCAM+ cells was significantly higher in patients with cancer than in healthy volunteers. Among the low-risk patients (n=9), two had up-staging and six had up-grading. Among those up-staged, there was one case which was EpCAM Among those cases up-graded, three were EpCAM In those with stage T2 tumors, the presence of Gleason pattern 5 was positively correlated with EpCAM positivity (rho=0.59, p<0.001). CTC counts in localized prostate cancer were associated with Gleason pattern 5. Active treatment should be considered for patients with low-risk disease during active surveillance who are found to have EpCAM+ CTCs because of a risk of up-staging and up-grading.

[This retracts the article DOI: 10.3892/ol.2018.8480.].

Analysis of anaplastic lymphoma kinase (ALK) rearrangement in non-small cell lung cancer (NSCLC) is considered to be a useful tool when considering predictive biomarker detection for evaluating eligibility for targeted therapy. It is not always possible to perform a tumor biopsy in patients. Isolation and culturing of circulating tumor cells (CTCs) may be an alternative to tumor biopsies for the diagnosis of ALK rearrangement. Blood was collected from 22 patients with NSCLC harboring ALK rearrangement and was divided into two groups: One for immunofluorescence staining and the other for culture. Samples were filtered by size and cultured CTCs were analyzed for echinoderm microtubule-associated protein-like 4-ALK translocation using fluorescence in situ hybridization. CTCs positive for epithelial cell adhesion molecule and CTCs exhibiting ALK rearrangement were detected. Therefore, CTCs may be used as a potential alternative method to tissue biopsy for diagnosing ALK rearrangement. Additionally, this method may have clinical applications including serial blood sampling for the development of personalized cancer therapy based on individual genomic information.

Liquid biopsy using circulating tumor cells (CTCs) is a noninvasive and repeatable procedure, and is therefore useful for molecular assays. However, the rarity of CTCs remains a challenge. To overcome this issue, our group developed a novel technology for the isolation of CTCs on the basis of cell size difference. The present study isolated CTCs from patients with breast cancer using this method, and then used these cells for cancer gene panel analysis. Blood samples from eight patients with breast cancer were collected, and CTCs were enriched using size-based filtration. Enriched CTCs were counted using immunofluorescent staining with an epithelial cell adhesion molecule (EpCAM) and CD45 antibodies. CTC genomic DNA was extracted, amplified, and screened for mutations in 400 genes using the Ion AmpliSeq Comprehensive Cancer Panel. White blood cells (WBCs) from the same patient served as a negative control, and mutations in CTCs and WBCs were compared. EpCAM+ cells were detected in seven out of eight patients, and the average number of EpCAM+ cells was 8.6. The average amount of amplified DNA was 32.7 µg, and the percentage of reads mapped to any targeted region relative to all reads mapped to the reference was 98.6%. The detection rate of CTC-specific mutations was 62.5%. The CTC-specific mutations were enhancer of zeste polycomb repressive complex 2 subunit, notch 1, AT-rich interaction domain 1A, serine/threonine kinase 11, fms related tyrosine kinase 3, MYCN proto-oncogene, bHLH transcription factor, APC, WNT signaling pathway regulator, and phosphatase and tensin homolog. The technique used by the present study was demonstrated to be effective at isolating CTCs at a sufficiently high purity for genomic analysis, and supported the use of comprehensive cancer panel analysis as a potential application for precision medicine.