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SummaryThe maintenance of genome integrity requires that telomerase action be limited to telomeres and not convert DSBs into neotelomeres. Using the breakpoint sequence from an apparent germline neotelomere formation event, we developed an assay to detect and quantify telomeric repeat addition at Cas9-programmed DSBs in human cells. The data show that telomerase can add telomeric repeats to DSBs and that this process can generate functional neotelomeres. Neotelomere formation is increased when telomerase is overexpressed, suggesting that in most human cells, low (or absent) telomerase activity limits the deleterious effects of de novo telomere addition. We show that neotelomere formation at DSBs is inhibited by long-range resection and the accompanying activation of ATR signaling. Our findings reveal that telomerase can cause genome instability by generating neotelomeres at DSBs. We propose that neotelomere formation can promote tumorigenesis by ending detrimental breakage-fusion-bridge cycles in cancer cells whose genome alterations engender dicentric chromosomes.

TRF1 facilitates the replication of telomeric DNA in part by recruiting the BLM helicase, which can resolve G-quadruplexes on the lagging-strand template. Lagging-strand telomeres lacking TRF1 or BLM form fragile telomeres—structures that resemble common fragile sites (CFSs)—but how they are formed is not known. We report that analogous to CFSs, fragile telomeres in BLM-deficient cells involved double-strand break (DSB) formation, in this case by the SLX4/SLX1 nuclease. The DSBs were repaired by POLD3/POLD4-dependent break-induced replication (BIR), resulting in fragile telomeres containing conservatively replicated DNA. BIR also promoted fragile telomere formation in cells with FokI-induced telomeric DSBs and in alternative lengthening of telomeres (ALT) cells, which have spontaneous telomeric damage. BIR of telomeric DSBs competed with PARP1-, LIG3-, and XPF-dependent alternative nonhomologous end joining (alt-NHEJ), which did not generate fragile telomeres. Collectively, these findings indicate that fragile telomeres can arise from BIR-mediated repair of telomeric DSBs.

Background Several studies showed that first-line bevacizumab plus chemotherapy for HER2-negative metastatic breast cancer improves progression-free survival and tumor response rate but not overall survival. MERiDiAN trial evaluated plasma vascular endothelial growth factor-A (VEGF-A) prospectively as a predictive biomarker for bevacizumab efficacy in metastatic breast cancer. However, results of this trial do not support using baseline plasma VEGF-A to identify patients benefitting most from bevacizumab. We measured baseline serum VEGF-A level from stored blood samples of metastatic breast cancer patient with treated bevacizumab plus paclitaxel as fist-line and later line therapy, and evaluated a correlation between serum VEGF-A level and efficacy of bevacizumab and prognosis of breast cancer patients tread with bevacizumab, retrospectively. Patients and methods We examined blood samples from 57 metastatic breast cancer patients treated with bevacizumab and paclitaxel, after obtaining written informed consent. And, we evaluated a correlation between baseline serum VEGF-A level and time to treatment failure (TTF) and overall survival (OS). We also compared the serum VEGF-A level of response group (CR and PR) and that of non-response group (SD and PD). Results Baseline serum level of VEGF-A ranged from 80 to 2079 pg/ml. Cases of treatment line were as follows: first-line, 22 cases (38.6%); second line, 11 cases (19.3%) and third-line and the later line, 24 cases (42.1%). The cutoff identified by ROC curve analysis that was able to differentiate response group and non-response group in first-line setting was 360pg/ml for serum VEGF-A. And, we separated high serum VGEF-A group and low serum VEGF-A group of patients treated with bevacizumab plus paclitaxel. In patients treated as first line therapy, median TTF was 4.0 months with high serum VGEF-A group versus 5.0 months with low serum VEGF-A group, and median OS was 12 months with high serum VGEF-A group versus 11months with low serum VEGF-A group. There were no significant differences in both TTF and OS in first line setting. In patients treated as second line and later line therapy, median TTF was 2.8 months with high serum VGEF-A group versus 7.1 months with low serum VEGF-A group, and median OS was 6.4 months with high serum VGEF-A group versus 12.7 months with low serum VEGF-A group. The prognosis of high serum VEGF-A group was significantly worse than that of low serum group in both TTF and OS. The serum VEGF-A level of response group was tend to be higher than that of non-response group in first line setting, and was lower in second and later line setting. However, there were no significant differences. Conclusion In this study, serum VEGF-A cannot be a predictor for efficacy of bevacizumab plus paclitaxel as first line therapy for metastatic breast cancer patients. On the other hand, there was a possibility that high serum level of VEGF-A can be a poor prognostic factor in late line therapy setting of bevacizumab. Citation Format: Tozuka K, Nagai SE, Matsumoto H, Hayashi Y, Kubo K, Tsuboi M, Sato A, Takai K, Wang X, Yamada Y, Inoue K. Prognostic and predictive value of serum level of vascular endothelial growth factor-A in metastatic breast cancer patients treated with bevacizumab plus paclitaxel [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 P5-12-17.

Telomere crisis contributes to cancer genome evolution, yet only a subset of cancers display breakage-fusion-bridge (BFB) cycles and chromothripsis, hallmarks of experimental telomere crisis identified in previous studies. We examine the spectrum of structural variants (SVs) instigated by natural telomere crisis. Eight spontaneous post-crisis clones did not show prominent patterns of BFB cycles or chromothripsis. Their crisis-induced genome rearrangements varied from infrequent simple SVs to more frequent and complex SVs. In contrast, BFB cycles and chromothripsis occurred in MRC5 fibroblast clones that escaped telomere crisis after CRISPR-controlled telomerase activation. This system revealed convergent evolutionary lineages altering one allele of chromosome 12p, where a short telomere likely predisposed to fusion. Remarkably, the 12p chromothripsis and BFB events were stabilized by independent fusions to chromosome 21. The data establish that telomere crisis can generate a wide spectrum of SVs implying that a lack of BFB patterns and chromothripsis in cancer genomes does not indicate absence of past telomere crisis., Telomere crisis has been shown to induce chromothripsis and breakage fusion bridge (BFB) cycles in vitro. Here, the authors show that telomere crisis generates a much broader spectrum of structural variations, implying that cancers without chromothripsis and BFB cycles could have emerged from telomere crisis.

Background Catheter ablation for atrial fibrillation (AF) with high contact force (CF) has been reported to lead to the gaps after pulmonary vein isolation (PVI), and the appearance of the gaps depends on the site in the left atrium (LA). Purpose The aim of this study is to clarify the relationship between the gaps appearance and high CF. Methods In the clinical study, 74 consecutive patients (39 males, age 71±9, 46 with paroxysmal AF) underwent conventional PVI with a point-by-point radiofrequency (RF) ablation using a CF sensing catheter (TactiCath SETM). RF energy (J), number of RF point, Lesion index (LSI), force-time integral (FTI), average CF (g), and the degree of LA depression with high CF were evaluated. In the experimental study (figure), the relationship between the ablation area and the degree of depression with high CF using bovine left ventricular wall. Results In the clinical study, the gaps were found in 175 of 1182 sites after first path PVI ablation. The CF in the gap sites was higher than without the gap sites in RPV roof, anterior RSPV and anterior RPV carina (18.9±6.8 vs 15.0±5.7g p=0.0262, 26.4±11.6 vs 19.5±6.0g p=0.0029, 22.7±6.4 vs 19.1±5.5g, p=0.0161). Meanwhile, there were no differences in RF energy, number of RF point, LSI, FTI. High CF (≥30g) showed a depression of 3 mm or more in only RPV roof and anterior RPV (figure). In the experimental study, the ablation range narrowed as the indentation deepened with more than 30g CF (r=0.6417, p=0.0625). Conclusion Catheter ablation for AF with high contact force might lead to the gaps in RPV roof and anterior RPV site by the reduction of the ablation area due to depression caused by the pressure. Extension of each contact force Funding Acknowledgement Type of funding source: None

Telomere shortening is a presumed tumor suppressor pathway that imposes a proliferative barrier (the Hayflick limit) during tumorigenesis. This model predicts that excessively long somatic telomeres predispose to cancer. Here, we describe cancer-prone families with two unique TINF2 mutations that truncate TIN2, a shelterin subunit that controls telomere length. Patient lymphocyte telomeres were unusually long. We show that the truncated TIN2 proteins do not localize to telomeres, suggesting that the mutations create loss-of-function alleles. Heterozygous knock-in of the mutations or deletion of one copy of TINF2 resulted in excessive telomere elongation in clonal lines, indicating that TINF2 is haploinsufficient for telomere length control. In contrast, telomere protection and genome stability were maintained in all heterozygous clones. The data establish that the TINF2 truncations predispose to a tumor syndrome. We conclude that TINF2 acts as a haploinsufficient tumor suppressor that limits telomere length to ensure a timely Hayflick limit.

In DNA repair, the resection of double-strand breaks dictates the choice between homology-directed repair—which requires a 3′ overhang—and classical non-homologous end joining, which can join unresected ends1,2. BRCA1-mutant cancers show minimal resection of double-strand breaks, which renders them deficient in homology-directed repair and sensitive to inhibitors of poly(ADP-ribose) polymerase 1 (PARP1)3–8. When BRCA1 is absent, the resection of double-strand breaks is thought to be prevented by 53BP1, RIF1 and the REV7–SHLD1–SHLD2–SHLD3 (shieldin) complex, and loss of these factors diminishes sensitivity to PARP1 inhibitors4,6–9. Here we address the mechanism by which 53BP1–RIF1–shieldin regulates the generation of recombinogenic 3′ overhangs. We report that CTC1–STN1–TEN1 (CST)10, a complex similar to replication protein A that functions as an accessory factor of polymerase-α (Polα)–primase11, is a downstream effector in the 53BP1 pathway. CST interacts with shieldin and localizes with Polα to sites of DNA damage in a 53BP1- and shieldin-dependent manner. As with loss of 53BP1, RIF1 or shieldin, the depletion of CST leads to increased resection. In BRCA1-deficient cells, CST blocks RAD51 loading and promotes the efficacy of PARP1 inhibitors. In addition, Polα inhibition diminishes the effect of PARP1 inhibitors. These data suggest that CST–Polα-mediated fill-in helps to control the repair of double-strand breaks by 53BP1, RIF1 and shieldin.

Background: To avoid tissue expander exposure following mastectomy flap necrosis, several methods for covering expander by autologous flap in cases of immediate breast reconstruction have been reported. These methods are classified into two groups, complete or partial expander coverage. Two methods have potential risks of postoperative complications following: insufficient lower pole expansion and cranial migration in complete coverage methods, and lateral migration in partial coverage methods. However, the comparisons of complication rates between these two methods have not been reported. This study aims to compare the incidence of expander exposure following mastectomy flap necrosis and expander migration between two methods. Methods: A retrospective review of 93 patients (99 breasts) who underwent immediate expander-based breast reconstruction was performed. Patients were divided into two groups, complete or partial expander coverage by autologous flaps. In both groups, expanders were placed into subpectral position. In partial coverage group, the lateral borders of pectralis major muscles were sutured to the mastectomy skin flaps. If the skin flap was too thin to be sutured, the serratus anterior muscro-fascial flap was dissected and sutured to the lateral border of pectralis major muscle to cover the expander completely. Allograft products were not used in both groups. Demographics, intraoperative findings, and postoperative complications were compared between two groups. Results: Of the 99 breasts, 56 underwent complete expander coverage and 43 underwent partial coverage. Mastectomy flap necrosis rate was higher in the complete coverage group (Complete 14.3% versus Partial 0%; p=0.0091), however, there was no incidence of expander exposure in both groups. Lateral migration rate was higher in the partial coverage group (Complete 0% versus Partial 9.3%; p=0.033). There was no difference in cranial migration rate between two groups (Complete 12.5% versus Partial 2.3%; p=0.133). Conclusions: The thinness of the mastectomy flaps was considered to provide the higher incidence of mastectomy flap necrosis in the complete coverage group. The complete expander coverage reduced lateral migration rate and prevented expander exposure in cases of mastectomy flap necrosis. Citation Format: Kubo K, Hamahata A, Tozuka K, Tsuboi M, Hayashi Y, Takai K, Saito T, Sakurai H, Matsumoto H. A complication analysis between complete and partial tissue expander coverage using autologous flaps in cases of immediate breast reconstruction [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P4-13-11.

Purpose TAFRO syndrome is a novel disorder manifesting as fever, anasarca, thrombocytopenia, renal insufficiency and organomegaly, and its etiology has not been clarified. The aim of this study was to elucidate similarities and differences between systemic lupus erythematosus (SLE) and TAFRO syndrome. Methods We examined 46 consecutive patients diagnosed with SLE and determined whether they meet the proposed diagnostic criteria for TAFRO syndrome (2015 version). Results Of the 46 patients with SLE, four (8.7%) also met the TAFRO syndrome criteria (TAFRO-like group). All patients in the TAFRO-like group were males, and their mean age was significantly higher than that of the non-TAFRO group (67.5 ± 8.7 vs. 39.3 ± 18.1 years, p = 0.004). C-reactive protein and γ-glutamyl transpeptidase levels were significantly higher, and frequencies of anti-dsDNA and anti-Sm antibodies were significantly lower in the TAFRO-like than non-TAFRO group. Elder cases (onset age ≥ 50 years) met significantly more categories of the diagnostic criteria for TAFRO syndrome than did those with younger cases. Conclusions Several patients with SLE, especially elder cases, showed features similar to those of TAFRO syndrome. Although exclusion of SLE is needed in the diagnostic criteria for TAFRO syndrome, TAFRO syndrome-like SLE should be considered.

Studies to date have not completely determined the factors influencing hydrogen embrittlement of ferrite/bainite X80 pipeline steel. Hydrogen embrittlement susceptibility was evaluated based on fracture strain in tensile testing. We conducted a thermal desorption analysis to measure the amount of tracer hydrogen corresponding to that of lattice defects. Hydrogen embrittlement susceptibility and the amount of tracer hydrogen significantly increased with decreasing crosshead speed. Additionally, a significant increase in the formation of hydrogen-enhanced strain-induced lattice defects was observed immediately before the final fracture. In contrast to hydrogen-free specimens, the fracture surface of the hydrogen-charged specimens exhibited shallower dimples without nuclei, such as secondary phase particles. These findings indicate that the presence of hydrogen enhanced the formation of lattice defects, particularly just prior to the occurrence of final fracture. This in turn enhanced the formation of shallower dimples, thereby potentially causing premature fracture of X80 pipeline steel at lower crosshead speeds.

Background: Neoadjuvant chemotherapy (NAC) with taxanes, followed by fluorouracil, epirubicin, and cyclophosphamide (FEC), with concurrent trastuzumab is known to achieve a high pCR rate of more than 60% for HER2-positive breast cancer (BC) as well as good prognoses in those obtaining pCR. On the other hand, the prognostic significance of tumor-infiltrating lymphocytes (TILs) has recently been described in triple-negative BC. However, the prognostic and predictive values of TILs in HER2-positive BC remain unclear. In the present study, we examined the grades of TILs in pre-treatment cancer tissues and residual tumors after NAC with trastuzumab, and also investigated its predictive utility for pCR and prognostic power for HER2-positive BC. Patients and Methods: A total of 128 Japanese women with HER2-positive BC received either paclitaxel or docetaxel followed by FEC, with concomitant trastuzumab. The proportional grades of stromal (Str)-TILs in pre-treatment biopsy specimens and residual tumors after NAC with trastuzumab were determined as follows: low grade (0-10%), intermediate grade (10-40%), and high grade (40-90%), using the criteria of the International Working Group for TILs in BC. Analysis 1: The relationship between the grades of Str-TILs in pre-treatment tumors and pCR rates was investigated. Relapse-free survival (RFS) and cancer-specific survival (CSS) were analyzed for a correlation with pre-treatment Str-TILs. Analysis 2: Alterations in the grade of Str-TILs were examined in the residual tumors of non-pCR patients, and RFS and CSS were analyzed for a correlation with residual Str-TILs. Results: pCR was achieved in 83 out of the 128 patients (pCR rate, 64.8%) who received NAC with trastuzumab, and RFS was significantly better in the pCR group than in the non-pCR group (p = 0.0071). Analysis 1: The patient distribution of the Str-TILs grade in pre-treatment tumors was as follows: high: 24 (18.8%); intermediate: 38 (29.7%); and low: 66 (51.6%). pCR rates correlated with the Str-TILs grade in pre-treatment tumors: 83.3% in the high group, 71.1% in the intermediate group, and 54.5% in the low group (p = 0.026); however, the Str-TILs grade in pre-treatment tumors did not correlate with survival. Analysis 2: In 45 non-pCR patients, the distribution of the Str-TILs grade in residual tumors was as follows: high: 9 (20.0%); intermediate: 8 (17.8%); and low: 28 (62.2%), respectively. In non-pCR patients, the rate of a high Str-TILs grade was greater in residual tumors than in pre-treatment tumors (residual, 20.0%, pre-treatment, 8.9%). RFS was significantly better with a high grade than with a low grade of residual Str-TILs (p = 0.033). Conclusions: The status of TILs in pre-treatment tumors predicted responses to NAC concomitant with trastuzumab in HER2-positive BC. The grade of TILs was higher in residual tumors than in pre-treatment tumors, and, among non-pCR patients, the prognosis of patients with a high residual-TILs grade was better prognosis than that of patients with a low residual-TILs grade. We speculate that an examination of TILs in residual tumors after NAC with trastuzumab may be necessary for selecting patients with a good prognosis from non-pCR patients. Citation Format: Kurozumi S, Inoue K, Matsumoto H, Hayashi Y, Tozuka K, Kubo K, Komatsu K, Takai K, Nagai SE, Oba H, Horiguchi J, Takeyoshi I, Kurosumi M. Prognostic value of tumor-infiltrating lymphocytes in residual tumors after neoadjuvant chemotherapy concomitant with trastuzumab for HER2-positive breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P4-14-17.

The power output of a wind turbine depends on both wind speed and turbine control. There is a fluctuation in the natural wind speed, which changes the turbine power output. In this paper blade pitch control based on detailed monitoring of the inflow wind, including vertical velocity distributions, has been experimentally evaluated in a field test wind turbine. The inflow wind speeds were observed by 11 sets of ultrasonic anemometers. In this first test, the blade pitch is changed to keep optimum angle of attack based on the local inflow wind speed in the rotor plane. The blade pitch is independently controlled as a reference radial position of 80% of rotor radius. As a result of the pitch controls, the wind turbine power output was increased and the power output fluctuation was decreased for the cyclic pitch control based on the local wind speed. With the cyclic pitch control, the fluctuation in rotor thrust and average rotor thrust were also reduced compere to collective pitch control.

Aim To assess detailed computed tomography (CT) findings in patients with the recently described thrombocytopenia, anasarca, fever, reticulin fibrosis, and organomegaly (TAFRO) syndrome, in order to contribute to imaging interpretation in the challenging diagnosis of this disease. Materials and methods The institutional review board approved this retrospective study and waived the need for informed consent. Eleven patients (six men, five women; mean age, 52.5 years) with confirmed TAFRO syndrome were included in this study. Chest-to-pelvis CT images were analysed for the presence of anasarca, organomegaly, bone lesions, and lung lesions. Results Anasarca was present in all patients and involved multiple cavities and tissues; pleural effusion and ascites were found in 100% of patients; pericardial effusion in 64%; periportal collar in 91%; gallbladder wall oedema in 78%; subcutaneous oedema in 91%; retroperitoneal oedema in 100%; and mesenteric oedema in 100%. Organomegaly involved multiple organs: hepatomegaly in 73%, splenomegaly in 82%, lymphadenopathy in 100%, and enlarged anterior mediastinum in 64% (solitary, well-circumscribed mass, 0%; infiltrative mass, 0%; non-mass-forming infiltrative lesion, 64%). Bone lesions were present in 91% patients and all bone lesions had ground-glass density with diffuse distribution. None of the patients had any lesions in their lungs. Conclusion The present study revealed that the findings of anasarca, organomegaly, and diffuse bony ground-glass appearance were observed in detail on CT in patients with TAFRO syndrome. A “matted” appearance of the enlarged anterior mediastinum is the characteristic CT finding of TAFRO syndrome, and it is possible to diagnose TAFRO syndrome from the combination of several CT findings.

Clarifying the states of hydrogen present in iron and steel is important in order to understand hydrogen embrittlement mechanisms and develop materials with high resistance to hydrogen embrittlement. Although it is widely recognized that the fracture strain of iron and steel decreases with increasing amounts of absorbed hydrogen, it is not yet well understood whether hydrogen directly decreases the fracture strain. Therefore, the objective of this study is to clarify the atomic-scale changes in strained α-iron specimens containing hydrogen. Low temperature thermal desorption spectroscopy (L-TDS), which can heat samples from lower temperatures than conventional TDS, was used to identify the peak temperatures and hydrogen states corresponding to various lattice defects in α-iron. The results indicate that new hydrogen trap sites in strained α-iron specimens containing hydrogen are enhanced compared to those without hydrogen. These sites are not dislocations, but hydrogen-enhanced strain-induced vacancies, because they are removed during aging at 30 °C.

The hydrogen-releasing properties from weld metals that mainly consist of martensite and contain some amount of retained austenite were experimentally investigated in order to determine the physical properties such as the activation energies for the simulation of the hydrogen distribution in welded joints and to consider the time period to age a cold cracking test specimen before crack observation. The numerical calculations for single-pass welding show that in the case of the 2.5 % and 4.2 % retained austenite, the hydrogen contents in the martensite become sufficiently low in 2 days. In the case of 16 % retained austenite, about 40 % of the initial hydrogen content still exists in the retained austenite 7 days after the completion of the welding. But the hydrogen content in the martensite becomes less than 1 % of the initial content in 7 days. Hence, the 7-day-ageing time before crack observation is considered to be adequate. The numerical calculations for multi-pass welded joints show that the 7-day ageing is also considered to be appropriate.

To prevent ATR activation, telomeres deploy the single-stranded DNA binding activity of TPP1/POT1a. POT1a blocks the binding of RPA to telomeres, suggesting that ATR is repressed through RPA exclusion. However, comparison of the DNA binding affinities and abundance of TPP1/POT1a and RPA indicates that TPP1/POT1a by itself is unlikely to exclude RPA. We therefore analyzed the central shelterin protein TIN2, which links TPP1/POT1a (and POT1b) to TRF1 and TRF2 on the double-stranded telomeric DNA. Upon TIN2 deletion, telomeres lost TPP1/POT1a, accumulated RPA, elicited an ATR signal, and showed all other phenotypes of POT1a/b deletion. TIN2 also affected the TRF2-dependent repression of ATM kinase signaling but not to TRF2-mediated inhibition of telomere fusions. Thus, while TIN2 has a minor contribution to the repression of ATM by TRF2, its major role is to stabilize TPP1/POT1a on the ss telomeric DNA, thereby allowing effective exclusion of RPA and repression of ATR signaling.

Background The detection of circulating tumor cells (CTCs) in peripheral blood is an independent predictor of the efficacy of systemic therapy, and also a prognostic marker for patients with metastatic breast cancer. One of the main methods to detect CTCs is CellSearch system, which uses immune-magnetic separation followed by immunocytochemistry. A microdevice (CTChip from ClearCell system) can capture and enumerate CTCs based on distinctive physiological differences (size and deformability) between cancer cells and blood cells. CTChip thus obtains a larger CTC yield than affinity-based separation, which enriches a particular subgroup of cells expressing EpCAM. In this study, we enumerate CTCs in peripheral blood from early and metastatic breast cancer patients using a size-based method. Patients and methods We examined blood samples from a total of 18 early and metastatic breast cancer patients, after obtaining written informed consent. Blood samples were taken in sodium EDTA tubes after discarding the first 1ml of blood from the syringe. Two ml blood samples were applied to CTChip (ClearCell system), and CTCs were eventually trapped in the microwells of the CTChip. Trapped cells were analyzed by immunocytochemistry with monoclonal antibodies specific for leukocytes (CD45) and epithelial cells (CK8/18), along with 4',6-diamidino-2-phenylindole (DAPI) for nuclei: CK8/18-positive, DAPI-positive and CD45-negative cells more than 10 μm in diameter were defined as CTCs. Eight patients were examined using both the CTChip and CellSearch system to compare the yield of CTCs. Results Of 18 patients, 6 were de novo stage IV, 6 were recurrent and 6 were early stage breast cancer patients. Of primary tumors, 8 were HER2- and ER and/or PR +, 6 were HER2-and ER- and PR-, 3 were HER2+ and ER and/or PR +, and one was HER2+ and ER- and PR-. Using CTChip, detected CTCs ranged from 3 - 107 cells/2 ml in all cases: 3 - 83 for early stage, 19 - 156 for stage IV and 21 - 146 for recurrent. The number of CTCs found in recurrent patients tended to be higher than in early stage patients. Size-based method using CTChip clearly showed high sensitivity compared with the CellSearch system, which detected CTCs in only 2 cases out of 8. In analysis by immunochemistry, we found CK-negative, CD45-negative and DAPI positive cells with larger diameter (>16 μm) than CK-positive CTCs in most patients, and the numbers were higher in stage IV (8.5 cells of median value) and recurrent (13 cells) patients than in early stage patients (1.5 cells). Our study suggested that CK-negative large cells might be CTCs with epithelial–mesenchymal transition (EMT). Conclusion This size-based technology enables us to capture CTCs regardless of EpCAM expression. Enumerated CTCs varied in size and positivity of CK8/18, suggesting the heterogeneity of CTCs. Further research, especially focusing on EMT will be crucial to understand the key mechanism of metastasis and drug resistance. Citation Format: Tozuka K, Nagai SE, Kubo K, Komatsu K, Takai K, Inoue K, Matsumoto H, Hayashi Y, Tsuboi M, Yamada Y, Wang X, Suganuma M. Enumeration of heterogeneous circulating tumor cells (CTCs) using size-based method in early, and metastatic, breast cancer patients [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P2-01-08.

The magnetoresistance (MR) properties in both the sputter-deposited Co/Si multilayers and the system consisting of Co evaporated on the anodized Si have been examined. In the Co/Si multilayers, at room temperature both the sharp ordinary magnetoresistance (OMR) and the negative granular-type giant magnetoresistance (GMR) appear, while at low temperatures only the large OMR of about 3.5% is observed for in-plane field. In the Co/anodized-Si system, at room temperature the MR is negligibly small, while it increases steeply with decreasing temperature and very large OMR of about 22% is obtained at 110 K for perpendicular field.

High-sensitive differential phase imaging has been performed with hard x-ray scanning transmission microscope optics and an imaging detector. A microbeam with the size of 60 nm at 8 keV x ray is generated by highly-coherent illuminated beam at x-ray undulalor beamline BL20XU of SPring-8. The differential phase image is reconstructed by data set consists of diffraction images of each scan position. Advantages for absorption imaging are shown with some objects.

Thermal desorption spectroscopy (TDS) has been used to reveal the nature of defects acting as trapping sites of hydrogen. Hydrogen was charged to ferritic and eutectoid steels deformed to various degrees and then given annealing treatment. Desorption with a single peak appeared between room temperature and 600 K from ferritic steels. Under constant hydrogen charging conditions, the amount of desorption increased with strain. However, when the deformed samples were subjected to annealing at temperatures as low as 500 K, the increase of desorbed hydrogen no longer appeared. Vacancy clusters, which themselves annihilate in the course of TDS measurement, are the probable source of hydrogen desorption. When heavy deformation was given to ferritic steels, a two-step decrease of hydrogen desorption took place with increasing annealing temperature, corresponding to annihilation of vacancy clusters and decrease of dislocation density, respectively. The desorption with a single peak has two origins, one due to the annihilation of the trapping sites themselves and the other to desorption from stable sites. For heavily deformed eutectoid steel, an additional desorption peak centered at around 640 K appeared. The peak likely results from deformation-induced defects within the cementite phase or supersaturated carbon in ferrite. Two types of desorption, one due to the annihilation of trapping sites in the course of measurement and the other due to desorption from stable sites, should be discriminated. TDS using hydrogen as a tracer can be applied as a tool to investigate the various defects induced by plastic deformation.

We report a method of endoscopic retrograde sphincterotomy in patients in whom the optimal viewing of the papilla of Vater is hardly obtained. Percutaneous transhepatic cholangiodrainage (PTCD)-tube is placed under ultrasonographic guidance. PTCD-tube coming out of the papilla of Vater is observed by the endoscope and the guide wire inside the shpincterotome is inserted into the PTCD-tube. Sphincterotome is advanced into the common bile duct by the guidance of the guide wire and PTCD-tube. Sphincterotomy is performed in a usual fashion. Two patients with previous history of gastrectomy underwent this procedure with successful results. This method should be tried when usual method of EST is difficult and unsuccessful.

The simultaneous measurement of both the relative electrical resistance and the equilibrium hydrogen and deuterium pressure as a function of composition of Pd - H and Pd - D systems have been carried out at temperatures between 273 and 323 K at pressures up to about 3.3 MPa. The relative resistance, , in the two-phase region for the absorption processes shows a very small and almost linear increase with increasing H(D) content, especially for the Pd - H system, compared to the larger changes previously observed by the electrolysis method. The resistance behaviour is quite similar to the shape of p - c isotherm relationships. The relative resistance increments per unit change of H(D)/Pd content at 298 K, , in the two-phase region are about 1.5 and 2.1 times larger for the Pd - H and Pd - D systems, respectively, compared to the changes in the relative lattice parameters with H(D)/Pd content, , within the two-phase region, where is the lattice parameter of H(D)-free Pd and r is the atom ratio. On the other hand, the resistance increment in the single solid solution phase and single phase, except for the higher-H(D)-content region, is significantly larger compared to the changes of the lattice expansion due to dissolved hydrogen and deuterium. Thus, the variation in resistance with hydrogen and deuterium content in the two-phase region may be mainly associated with an incoherent formation of hydride within the phase. The relative resistance for the subsequent desorption processes from the absorption up to about 3.3 MPa at 298 K in both Pd - H and Pd - D systems exhibits almost the same maximum as that of the absorption processes, i.e. at about and at about , and then the values decrease gradually with decreasing H(D) content up to the phase boundary composition; on entering the two-phase region, the values remain almost constant, i.e. for the Pd - H system and for the Pd - D system. This large hysteresis of resistance can be attributed to the creation of `lattice strain deformations' accompanied by dislocation formation from hydride (deuteride) formation and by further highly dissolved hydrogen and deuterium in the phase region.

The enthalpies for the reaction of gaseous hydrogen and deuterium with palladium have been measured as a function of [H]/[Pd] and [D]/[Pd] atomic ratios up to 0.865 and 0.85, respectively, in the temperature range 298 - 194.5 K using a differential-heat-flow low-temperature calorimeter; pressure - concentration isotherms have been measured simultaneously. The calorimetric two-phase enthalpies for the Pd - H and Pd - D systems at 298 K are and , and the corresponding entropies determined from the calorimetric enthalpies and the absorption plateau pressures are and . These - and -values are in agreement with the previously reported values and also with values determined in the literature from van't Hoff plots of the plateau pressures. The calorimetrically determined enthalpies for solution in the -phase regions of both systems become less exothermic in an almost linear fashion with increasing H(D) content, independently of temperature. At the same H(D) content, the -values for the Pd - H system are slightly more exothermic than the -values for the Pd - D system. Enthalpies of almost the same magnitude are obtained from the desorption data. The corresponding entropies, for solution in the -phase region of both systems have a tendency to decrease gradually with increase in H(D) content independently of temperature; however, there is no marked difference between the magnitudes of the -values of the two systems. The dependences of the calorimetric enthalpies for hydrogen (deuterium) solution on H and D contents in the ranges and can be expressed by the temperature-independent relations , and . The variations in -values with H(D) content are in agreement with that calculated from van't Hoff plots of the previously reported relative chemical potentials of hydrogen and deuterium at low temperatures. The present calorimetric data determined by gas-phase measurements do not offer support for the generation of any `excess heat' up to [D]/[Pd] = 0.85, beyond that which is expected from the chemical reaction because the values determined calorimetrically are almost the same as those derived from van't Hoff plots.

The enhancement of the ordinary magnetoresistance (OMR) in Si has been attempted by constructing two type of Co∕Si systems; one is the sputter-deposited current perpendicular to plane-type Co∕Si multilayers and the other is the sputtered-Co∕anodized-porous-Si system. The Co∕Si multilayers show the sharp OMR at a low field and the negative granular-type giant magnetoresistance at higher fields. At 50K, however, only the OMR appears with the ratio of about 3.0% for an in-plane field of 10kOe. In the Co∕anodized-Si system, a very large OMR of about 60% is observed at 50K for perpendicular field of 10kOe.

We report a 61-year-old woman with definite diagnosis of isolated neurosarcoidosis in the medulla oblongata involving the fourth ventricle. We could not recognize neurosarcoidosis as one of the differential diagnoses of the lesion before biopsy because the brainstem lesion location and periventricular lesion configuration were quite unusual.

Behçet's disease (BD) is a multi-systemic inflammatory disease, characterised by recurrent oral aphthosis, genital ulcers, skin lesions and uveitis. We have reported excessive Th1 cell activity in patients with BD. More recently, Th17 cells were suggested to associate with several autoimmune diseases. This study was designed to investigate the role of Th17 related cytokines and signalling molecules in patients with BD.We examined mRNA expressions of Th1 and Th17 related cytokines and related signalling molecules in PBMC of 12 patients with BD and 14 normal controls (NC) using quantitative RT-PCR. We studied expressions of the Th17 related cytokines in other four BD patients' skin lesions by immunofluorescence.Major Th17 related cytokines were not detected in unstimulated PBMC in patients with BD. After stimulation, mRNA expressions of TGFβ receptor type 1, IL-12 receptor β2 and suppressor of cytokine signalling protein (SOCS) 1 on PBMC were significantly enhanced in patients with BD, as compared with NC (p0.05). mRNA expression of RORC, a key transcription factor for Th17 cell differentiation, was comparable between BD and NC. CD4+ T cells infiltrating into BD skin lesion expressed TGFβ1 much more than those infiltrating into non-Behçet's disease erythema nodosum.These findings suggest that TGFβ/Smad signalling pathway of T cells is overactive in patients with BD.

Human telomeres bind shelterin, the six-subunit protein complex that protects chromosome ends from the DNA damage response and regulates telomere length maintenance by telomerase. We used quantitative immunoblotting to determine the abundance and stoichiometry of the shelterin proteins in the chromatin-bound protein fraction of human cells. The abundance of shelterin components was similar in primary and transformed cells and was not correlated with telomere length. The duplex telomeric DNA binding factors in shelterin, TRF1 and TRF2, were sufficiently abundant to cover all telomeric DNA in cells with short telomeres. The TPP1.POT1 heterodimer was present 50-100 copies/telomere, which is in excess of its single-stranded telomeric DNA binding sites, indicating that some of the TPP1.POT1 in shelterin is not associated with the single-stranded telomeric DNA. TRF2 and Rap1 were present at 1:1 stoichiometry as were TPP1 and POT1. The abundance of TIN2 was sufficient to allow each TRF1 and TRF2 to bind to TIN2. Remarkably, TPP1 and POT1 were approximately 10-fold less abundant than their TIN2 partner in shelterin, raising the question of what limits the accumulation of TPP1 x POT1 at telomeres. Finally, we report that a 10-fold reduction in TRF2 affects the regulation of telomere length but not the protection of telomeres in tumor cell lines.