Quantitative monitoring of circulating tumor DNA in patients with advanced pancreatic cancer undergoing chemotherapy.

According to cancer genome sequences, more than 90% of cases of pancreatic ductal adenocarcinoma (PDAC) harbor active KRAS mutations. Digital PCR (dPCR) enables accurate detection and quantification of rare mutations. We assessed the dynamics of circulating tumor DNA (ct-DNA) in patients with advanced PDAC undergoing chemotherapy using dPCR. KRAS G12/13 mutation was assayed by dPCR in 47 paired tissue- and ct-DNA samples. The 21 patients were subjected to quantitative ct-DNA monitoring at 4 to 8-week intervals during chemotherapy. KRAS mutation was detected in 45 of those 47 patients using tissue DNA. In the KRAS mutation-negative cases, next-generation sequencing revealed KRAS Q61K and NRAS Q61R mutations. KRAS mutation was detected in 23/45 cases using ct-DNA (liver or lung metastasis, 18/19; mutation allele frequency [MAF], 0.1%-31.7%; peritoneal metastasis, 3/9 [0.1%], locally advanced, 2/17 [0.1%-0.2%]). In the ct-DNA monitoring, the MAF value changed in concordance with the disease state. In the 6 locally advanced cases, KRAS mutation appeared concurrently with liver metastasis. Among the 6 cases with liver metastasis, KRAS mutation disappeared during the duration of stable disease or a partial response, and reappeared at the time of progressive disease. The median progression-free survival was longer in cases in which KRAS mutation disappeared after an initial course of chemotherapy than in those in which it was continuously detected (248.5 vs 50 days, P < .001). Therefore, ct-DNA monitoring enables continuous assessment of disease state and could have prognostic utility during chemotherapy.

Numerical simulations of partially coherent illumination for multi-scattering phase objects via a beam propagation method.

Many studies have performed imaging under partially coherent illumination. However, to the best of our knowledge, there are no imaging methods for complicated objects such as cell colonies, which are large and diffract light multiple times. In this paper, we propose an image calculation method for the partially coherent illumination of a large-scale three-dimensional multi-diffractive object. The image is calculated via the summation of coherent images of each illumination angle using the beam propagation method. We apply this method to various microscopic observations, including phase contrast and differential interference contrast. Our method shows excellent agreement with experimental images of a bead and a photonic crystal fiber. As with a cell colony, our method reproduced the characteristics of the image through experimentation. Finally, we discuss the accuracy and the restriction conditions of our method.

Spiroborate-Based Double-Stranded Helicates: Meso-to- Racemo Isomerization and Ion-Triggered Springlike Motion of the Racemo-Helicate.

A one-handed double-stranded spiroborate helicate exhibits a unique reversible extension-contraction motion coupled with a twisting motion in one direction triggered by binding and release of a Na+ ion while retaining its handedness. Here we report that an extended meso-helicate was also produced together with the racemo-helicate, and the meso-helicate was readily converted to the racemo-helicate assisted by a Na+ ion as a template in the presence of water. The thermodynamic analyses of the ion-triggered springlike motion of the racemo-helicate using a series of monovalent cations with different sizes revealed that the association constants of the extended racemo-helicate decreased in the following order: Li+ > Na+ > NH4+ > Ag+ ≥ K+ > Cs+ > Rb+, which roughly agrees with the reverse order of their ionic radii except for the NH4+ ion due to the more elongated contracted helicates when complexed with larger cations as supported by the crystal and DFT calculated structures. The one-handed contracted helicates showed characteristic CD spectra depending on the cation species due to the differences in their contracted helical structures, and its absolute handedness of the spiroborate helicate was determined by X-ray crystallography. The kinetic studies of the springlike motions of the racemo-helicate showed that the exchange rate between the extended and contracted helicates tend to increase in the following order: Li+ < Na+ < K+ ≤ NH4+ < Rb+ < Cs+ < Ag+ as anticipated from the association constants, being in good agreement with the order of the cation sizes except for Ag+.

Quantitative phase imaging by optimized asymmetric illumination.

We have presented a simple approach for quantitative phase imaging by optimizing asymmetric illumination of a conventional microscope. With this illumination, the light intensity modulation accompanying refraction at the surface profile of phase objects occurs, and "phase-gradient information" can be derived by detecting it. Two images with phase-gradient information on different axes are converted into the two-dimensional phase distribution of the specimen by introducing the phase-gradient transfer function, which is the intensity change due to refraction by the phase-gradient of a specimen. We experimentally confirm accurate and repeatable performance of our method and demonstrate phase imaging of live cells.

Allosteric Regulation of Unidirectional Spring-like Motion of Double-Stranded Helicates.

We report the unprecedented allosteric regulation of the extension and contraction motions of double-stranded spiroborate helicates composed of 4,4'-linked 2,2'-bipyridine (bpy) and its N,N'-dioxide units in the middle of ortho-linked tetraphenol strands. NMR and circular dichroism measurements and an X-ray crystallographic analysis along with theoretical calculations revealed that enantiomeric helicates contract and extend upon the binding and release of protons and/or metal ions at the covalently linked two binding bpy or N,N'-dioxide moieties without racemization, respectively, regulated by a cooperative anti-syn conformational change of the two bpy or N,N'-dioxide moieties. These anti-syn conformational changes that occurred at the linkages are amplified into a large-scale molecular motion of the helicates leading to reversible spring-like motions coupled with twisting in one direction in a highly homotropic allosteric fashion.

Method for observing phase objects without halos or directional shadows.

A new microscopy method for observing phase objects without halos or directional shadows is proposed. The method is based on transformation of the surface profile of phase objects into a light intensity pattern. The key optical element is an annular aperture at the front focal plane of a condenser. The light flux passing through the annular aperture is changed by the specimen's surface profile and then passes through an objective and contributes to image formation. Theory and experimental results are presented. Phase images of induced pluripotent stem (iPS) cells with clear outlines were obtained with this method.

Activation of STING in pancreatic cancer-associated fibroblasts exerts an antitumor effect by enhancing tumor immunity.

Pancreatic ductal adenocarcinoma (PDAC) has a high mortality rate; therefore, the development of effective treatments is a priority. The stimulator of interferon genes (STING) pathway enhances tumor immunity by inducing the production of type 1 interferon (IFN) and proinflammatory cytokines and chemokines and promoting the infiltration of cytotoxic T cells. To assess the function of STING on pancreatic tumorigenesis, Ptf1aER-Cre/+ LSL-KrasG12D/+ p53loxP/loxP mice (KPC mice) and Ptf1aER-Cre/+ LSL-KrasG12D/+ p53loxP/loxP/STING-/- mice (KPCS mice) were generated. However, STING deletion did not affect pancreatic tumorigenesis in mice. Because STING is expressed not only in immune cells but also in cancer-associated fibroblasts (CAFs), we evaluated the STING function in PDAC CAFs. A mouse STING agonist 5,6-Dimethyl-9-oxo-9H-xanthene-4-acetic acid (DMXAA) was administered to KPC mice and CAFs from KPC mice and the resulting immune response was evaluated. DMXAA activated STING in PDAC CAFs in KPC mice, promoting cytotoxic T cell infiltration by secreting proinflammatory cytokines and enhancing tumor immunity. We next generated STING-deficient PDAC cells and subcutaneous tumors in which STING was expressed only in CAFs by performing bone marrow transplantation and assessed the antitumor effect of STING-activated CAFs. The administration of DMXAA to subcutaneous tumors expressing STING only in CAFs sustained the antitumor effect of DMXAA. About half of human PDACs lacked STING expression in the cancer stroma, suggesting that STING activation in PDAC CAFs exerts an antitumor effect, and STING agonists can be more effective in tumors with high than in those with low STING expression in the stroma.

Clinical Significance of Serglycin Expression in Human Breast Cancer Patients.

BACKGROUND/AIM: Serglycin plays a crucial role in the aggressiveness of several types of malignancies, including breast cancer. In this study, we aimed to investigate the prognostic impact of serglycin expression in breast cancer patients, which has not been previously reported. PATIENTS AND METHODS: Immunohistochemical analyses were performed on 348 resected specimens of invasive carcinomas, using antibodies against serglycin. RESULTS: Low serglycin expression was observed in 23% of specimens (80/348) and significantly correlated with high histological grade (p=0.001) and negative ER (p=0.013). The log-rank test showed that low serglycin expression correlated with shorter distant metastasis-free survival (DMFS) (p=0.016) and disease-specific survival (DSS) (p=0.037) in node-positive breast cancer patients. Cox's multivariate analysis revealed that low serglycin expression was an independent factor for shorter DMFS (p=0.017) and DSS (p=0.020) in node-positive breast cancer patients. CONCLUSION: Low serglycin expression is an independent predictor of unfavorable prognosis in node-positive breast cancer patients.

EGFR inhibition reverses resistance to lenvatinib in hepatocellular carcinoma cells.

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related death worldwide. Lenvatinib is approved as a first-line treatment for unresectable HCC. The therapeutic duration of lenvatinib is limited by resistance, but the underlying mechanism is unclear. To establish lenvatinib-resistant cells, Hep3B cells were initially treated with 3 µM lenvatinib. The concentration was gradually increased by 1 µM or 0.5 µM per week and it reached to 7.5 µM 2 months after the initial exposure to lenvatinib. The biological characteristics of these cells were analyzed by ERK activation in the MAPK signaling pathway and a human phospho-receptor tyrosine kinase (RTK) antibody array. Factors possibly related to lenvatinib resistance were analyzed using inhibitors, and cell proliferation was analyzed. We established lenvatinib-resistant HCC cells (LR cells) by long-term exposure to lenvatinib. Lenvatinib reduced ERK activation in the parent cells, but not in the LR cells. RTK array analysis showed that the activities of EGFR and insulin-like growth factor 1 receptor (IGF1R)/insulin receptor (INSR) were significantly increased in LR cells, whereas the activities of other RTKs were unchanged. Erlotinib, a widely used EGFR inhibitor, downregulated ERK activation in LR cells. The proliferation of LR cells will also be affected when lenvatinib is combined with erlotinib to treat LR cells. In contrast, inhibition of IGFR/INSR did not affect ERK activation or cell proliferation. Scavenging of reactive oxygen species (ROS) ameliorated the enhanced EGFR activation in LR cells. Lenvatinib resistance was induced by enhanced EGFR activation, possibly via ROS accumulation, in lenvatinib- resistant cells. These findings may enable the development of lenvatinib combination therapies for HCC.

Podoplanin expression in cancer-associated fibroblasts predicts unfavorable prognosis in node-negative breast cancer patients with hormone receptor-positive/HER2 - negative subtype.

BACKGROUND: Podoplanin expression in cancer-associated fibroblasts (CAFs) has been proposed as an indicator for poor prognosis in patients with invasive breast carcinomas, but little is known about its clinical significance in node-negative breast cancer patients with hormone receptor (HR) + /HER2 - subtype, who are expected to have a favorable prognosis. METHODS: Immunohistochemical analyses were performed on 169 resected specimens of node-negative invasive carcinoma of no special type with HR + /HER2 - subtype using antibodies for podoplanin. When more than 10% of CAFs showed immunoreactivity with podoplanin as strong as that of internal positive controls, the specimens were judged as podoplanin-positive. RESULTS: Podoplanin-positive status in CAFs was observed in 16.0% (27 of 169 cases) and it associated with high Ki67 labeling index (LI) (> 30%) (p = 0.03), higher stromal tumor-infiltrating lymphocytes (p < 0.001) and progesterone receptor-negative status (p = 0.045). Log-rank test showed that podoplanin-positive status in CAFs correlated with shorter disease-free survival (DFS) (p = 0.007) and disease-specific survival (DSS) (p < 0.001). Univariate analysis showed a significant correlation between shorter DFS and podoplanin-positive status in CAFs (hazard ratio [HR] = 3.380; p = 0.012), the presence of lymphatic invasion (HR = 5.621; p < 0.001), high Ki67 LI (HR = 5.217; p < 0.001), and histological grade III (HR = 3.748; p = 0.008). According to Cox multivariate analysis, podoplanin-positive status in CAFs had the most significant effect on shorter DSS (HR = 37.759; p = 0.003) followed by high Ki67LI (HR = 27.664; p = 0.007). CONCLUSION: Podoplanin expression in CAFs could be an independent predictor for poor prognosis in node-negative breast cancer patients with HR + /HER2 - subtype.

Multiple-line Chemotherapy for a Patient with Unresectable Mucinous Cystic Neoplasm of the Pancreas.

A 74-year-old woman with a cyst in her pancreatic tail was referred to our hospital. Computed tomography confirmed a large cystic lesion with irregular wall thickening, abdominal lymph node swelling, and ascites. We diagnosed her with an unresectable mucinous cystic neoplasm, since ascites cytology revealed adenocarcinoma. The patient received chemotherapy up to the fifth line for 55.2 months. Gemcitabine plus nab-paclitaxel and modified FOLFIRINOX achieved a partial response with a progression-free survival time of 12.1 and 20.4 months, respectively. The overall survival time from the beginning of first-line chemotherapy was 69.4 months.

[Paroxetine treatment of 3 cases of cholestatic pruritus due to gastrointestinal malignancy].

Recently, the antipruritic activity of the selective serotonin reuptake inhibitor paroxetine has been reported. We report 3 patients with severe cholestatic pruritus due to gastrointestinal malignancy. All of them were successfully treated with 10 mg oral paroxetine. The onset of the antipruritic action of paroxetine was observed at from several hours-two days. Paroxetine seems to be an effective, well-tolerated treatment for cholestatic pruritus due to gastrointestinal malignancy.

Influence of irrigation water intake on local increase of radiocesium activity concentration in rice plants near a water inlet.

To identify the cause of the phenomenon that rice plants close to the water inlet contain relatively higher radiocesium within a paddy field plot, we conducted a field experiment by establishing experimental channel using polypropylene corrugated sheets, and sampling surface water, paddy soil and rice plants according to the distance from the water inlet in 2014 and 2015. It was found that the 137Cs activity concentrations in both dissolved and particulate forms in paddy surface water presented a declining trend from the water inlet towards the outlet. The 137Cs activity concentration in paddy soil in the harvesting season and those of brown rice and rice straws were highest at 1-2 m from the water inlet. Balance calculation suggests that destination of the lost 137Cs from the surface water was likely to be adsorption of the dissolved form and sedimentation of particulate form onto the soil. The concentration of exchangeable potassium ion in paddy soil was below the recommended standard of 250 mg kg-1 (as K2O in dry soil) near the water inlet at the harvesting period both years．These findings suggested that the possible crucial factors to induce rice plant uptake of radiocesium near the water inlet were either (1) direct absorption of dissolved 137Cs in surface water by rice plants, (2) absorption of 137Cs, which was originally retained in particulate matter and released by ion exchange and/or by organic matter decomposition in combination with (3) loss of soil exchangeable potassium caused due partly to transportation of soil particles with exchangeable potassium by the rapid water flow near the water inlet and/or leaching by ion exchange onto the soil of other cations such as calcium ion flowing into the paddy field. These findings will contribute to providing possible measures for producing safe rice in highly contaminated areas in which agricultural production will resume in the near future. We propose providing a non-planting zone for the area closer than about 5 m from the water intake to avoid the occurrence of high 137Cs concentrations in rice crops.

Fluorescent molecular spring that visualizes the extension and contraction motions of a double-stranded helicate bearing terminal pyrene units triggered by release and binding of alkali metal ions.

A novel double-stranded spiroborate helicate bearing terminal pyrene residues exhibited reversible fluorescence color changes (green and blue colors) due to the unique unidirectional springlike motions of the helicate triggered by catch and release of alkali metal ions.

Bronchoplastic lobectomy with wide wedge resection for lung cancer with long-term steroid medication.

A 57-year-old man with erythrodermia, who was given 5-10 mg/day of prednisolone for 2.5 years, was admitted to our hospital for squamous cell lung carcinoma of the right upper lobe. A bronchoscopy revealed a tumor nearly obstructing the right upper lobe bronchus. A bronchoplastic lobectomy was performed with wide wedge resection of the main bronchus and truncus intermedius. A postoperative bronchoscopy revealed good healing of the anastomosis and a 3-dimensional construction of the bronchus with chest computed tomography demonstrated no stenotic change and no kinking change in the anastomosis. One year and 6 months after surgery, no local recurrence was seen in the region of bronchoplasty. Bronchoplastic lobectomy with wide wedge resection is a useful procedure in cases with risk factors of anastomotic dehiscence, such as after induction therapy or during long-term administration of adrenal cortical steroids.

Prognostic significance of T helper 1 and 2 and T cytotoxic 1 and 2 cells in patients with non-small cell lung cancer.

BACKGROUND: We report the prognostic significance of peripheral and tumor-infiltrating Th1, Th2, Tc1 and Tc2 cells in lung cancer patients. MATERIALS AND METHODS: We evaluated the rates of interferon (IFN)-gamma+/CD4+ cells (Th1), interleukin (IL)-4+ /CD4+ cells (Th2), IFN-gamma+/CD8+ cells (Tc1), IL-4+ /CD8+ cells (Tc2), and the ratio of Th1 to Th2 and that of Tc1 to Tc2 among peripheral blood lymphocytes (PBL) and tumor-infiltrating lymphocytes (TIL), in 51 consecutive patients with non-small cell lung cancer, by detecting the intracellular cytokine production using three-color flow cytometry. RESULTS: Patients with a low Th1/Th2 ratio in peripheral blood lymphocytes had a significantly better prognosis than those with a high Th1/Th2 ratio (5-year survival rate: low: 74.7% vs. high: 50.3%; p=0.038). Patients with a low Th1/Th2 ratio in peripheral blood had a significantly better prognosis than those with a high Th1/Th2 ratio in pathological Stage II or III (5-year survival rate: low: 66.6% vs. high: 18.2%; p=0.018). CONCLUSION: A high Th1/Th2 ratio in peripheral blood is a negative prognostic factor, especially in pathological Stage II or III non-small cell lung cancer patients.

[Pathogenesis, diagnosis, and treatment of postoperative deep vein thrombosis].

Most postoperative deep vein thrombosis (DVT) arises from the venous systems of the pelvis and lower extremities, especially the soleal veins. Embolization of venous thrombi is related to the size and location of thrombi and movement of the lower limbs and commonly occurs within 1 week from the onset of formation. There are three steps in the final diagnosis of DVT: probable diagnosis by anamnesis and physical examination; screening diagnosis using quantitative tests; and definitive diagnosis using imaging tests. To determine embolic sources, venous echography, which is noninvasive and convenient, is the first choice. Therapeutic methods are selected based on thrombi extent and time after formation. Anticoagulant therapy is indicated in all cases except in patients with possible bleeding tendency and continues for 3 months or more. Among the endovascular therapies, catheter-directed thrombolysis is a more effective approach when combined with a temporary vena cava filter than operative thrombectomy. Although the prevention of DVT is mandatory for surgeons, it is difficult to avoid venous thromboembolism completely. Systemic early diagnosis and emergent therapeutic strategies for venous thromboembolism are clinically effective and promising.

Mucosa-associated Lymphoid Tissue Lymphoma of the Trachea in a Patient with Breast Cancer.

We herein report the case of a 93-year-old woman with breast cancer on the left side. Preoperative computed tomography of the chest showed irregularities and narrowing of the mid-trachea. Bronchoscopy was performed, and the results of a biopsy supported a diagnosis of mucosa-associated lymphoid tissue (MALT) lymphoma. The patient responded to treatment with prednisone alone, with a reduction in the size of the lesion. MALT lymphoma of the trachea is extremely rare, and there are only a few case reports of double cancer, i.e., MALT lymphoma of the trachea and breast cancer.

Hemoperfusion with polymyxin B-immobilized fiber in septic patients with methicillin-resistant Staphylococcus aureus-associated glomerulonephritis.

BACKGROUND/AIMS: We investigated whether urinary podocytes are present in septic patients with methicillin-resistant Staphylococcus aureus (MRSA)-associated glomerulonephritis and whether polymyxin B-immobilized fiber (PMX-F) treatment affects proteinuria and urinary podocyte excretion in these patients. METHODS: Twenty septic patients with MRSA-associated glomerulonephritis (mean age: 63.7 years) and 80 septic patients whose MRSA infection was not followed by glomerulonephritis (mean age: 60.5 years) were included in this study. All septic patients were treated with fosfomycin sodium, beta-lactams, arbekacin sulfate, and teicoplanin, or a combination of these. Twenty septic patients with MRSA-associated glomerulonephritis were randomly assigned to one of two treatments: PMX-F treatment (group A, n = 10) and conventional treatment (group B, n = 10). PMX-F treatment was repeated twice. RESULTS: Urinary podocytes and urinary protein excretion were not detected in MRSA septic patients without glomerulonephritis. However, urinary podocytes (1.7 +/- 0.6 cells/ml) and proteinuria (2.6 +/- 0.6 g/d) were detected in the 20 septic patients with MRSA-associated glomerulonephritis. Plasma endotoxin levels were decreased from 13.6 +/- 4.6 pg/ml to 6.6 +/- 2.2 pg/ml (p < 0.05) in group A. Levels in group B, however, showed little difference after treatment. Urinary podocytes were reduced in group A (from 1.8 +/- 0.6 cells/ml to 0.4 +/- 0.2 cells/ml, p < 0.01) as was urinary protein excretion (from 3.0 +/- 0.5 g/d to 0.8 +/- 0.4 g/d, p < 0.01) but urinary podocytes and protein excretion levels showed little difference after treatment in group B. CONCLUSION: PMX-F treatment may be effective in reducing urinary protein and urinary podocyte excretion in septic patients with MRSA-associated glomerulonephritis.

Hemoperfusion with polymyxin B immobilized fibers for urinary albumin excretion in septic patients with trauma.

We investigated whether microalbuminuria/urinary creatinine ratio (MACR) is increased in septic patients with trauma and whether polymyxin B immobilized fiber (PMX-F) treatment decreases MACR. Twelve trauma patients without sepsis, 18 trauma patients with sepsis, and 10 healthy controls were included in this study. The 18 trauma patients with sepsis were randomly assigned to one of two groups, PMX-F treatment or conventional treatment. Urinary microalbumin and creatinine were measured before and after treatment. Plasma endotoxin levels were determined by endospecy test. Hemoperfusion with PMX-F was carried out twice, for 2 hours, at a flow rate of 100 ml/min. MACR increased in the 30 trauma patients (5.2+/-2.2 mg/mmol) in comparison to that in the healthy controls (1.0+/-0.6 mg/mmol, p < 0.01). In the 18 trauma patients with sepsis, MACR after sepsis (16.6+/-4.8 mg/mmol) was significantly greater than that before sepsis (5.5+/-2.3 mg/mmol, p < 0.01). There was a significant correlation between plasma endotoxin levels and MACR in septic trauma patients (p < 0.001). MACR was reduced from 17.0+/-5.0 mg/mmol to 4.2+/-1.5 mg/mmol (p < 0.01) with PMX-F, and plasma endotoxin levels were also reduced from 34.5+/-18.5 pg/ml to 10.8+/-6.6 pg/ml (p < 0.01). Neither MACR nor plasma endotoxin levels were affected by conventional treatment, however. In summary, trauma patients with sepsis appear to show increased MACR, and PMX-F therapy may be effective for attenuating the increase in MACR.