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Prostate cancer (PCa) is a clinically heterogeneous disease. Predicting clinically significant PCa with low–intermediate prostate-specific antigen (PSA), which often includes aggressive cancers, is imperative. This study evaluated the predictive accuracy of deep learning analysis using multimodal medical data focused on clinically significant PCa in patients with PSA ≤ 20 ng/mL. Our cohort study included 178 consecutive patients who underwent ultrasound-guided prostate biopsy. Deep learning analyses were applied to predict clinically significant PCa. We generated receiver operating characteristic curves and calculated the corresponding area under the curve (AUC) to assess the prediction. The AUC of the integrated medical data using our multimodal deep learning approach was 0.878 (95% confidence interval [CI]: 0.772–0.984) in all patients without PSA restriction. Despite the reduced predictive ability of PSA when restricted to PSA ≤ 20 ng/mL (n = 122), the AUC was 0.862 (95% CI: 0.723–1.000), complemented by imaging data. In addition, we assessed clinical presentations and images belonging to representative false-negative and false-positive cases. Our multimodal deep learning approach assists physicians in determining treatment strategies by predicting clinically significant PCa in patients with PSA ≤ 20 ng/mL before biopsy, contributing to personalized medical workflows for PCa management.

Abstract Purpose The COVID-19 pandemic has affected lives and dental treatment. Aerosols and droplets generated during dental treatment present a risk of infection for dental care workers. However, detailed elucidation of the conditions under which those are generated has yet to be presented, and no clear countermeasures for protection have been established. The present study aimed to clarify the process of generation of aerosol and droplets in dental treatment, as well as their dynamics for establishment of effective countermeasures and protection strategies. Methods Images were obtained using a high-speed camera of aerosol and droplets generated during dental treatments performed on a mannequin. The effects of intraoral vacuum and extraoral vacuum to reduce those, as well as splash range with different body position were examined. Quantitative evaluations of aerosol and droplets were also performed using water-sensitive paper. Results Aerosol and droplets quantities were significantly reduced by use of both intraoral and extraoral vacuums as compared to no vacuum in both image analysis and findings obtained with water-sensitive paper (p

Abstract Background The presence of collateral vessels (CVs) on contrast-enhanced computed tomography is a poor prognostic factor in renal cell carcinoma (RCC), but its value in small RCC (sRCC;

Glass pH sensors are unsuitable for in vivo biomedical, clinical, or food applications because of brittleness of glass, the difficulty in measurement of small volumes and complex matrices, and the need for calibration. Herein, a novel enamel reference electrode for pH sensors is developed using a binary xAg2O·(100–x)TeO2 glass/stainless steel (SUS) electrode. Subsequently, the effects of Ag2O content on the precipitated crystalline phases and pH sensitivity of Ag2O–TeO2 glass/SUS electrodes are examined. The results indicate that the xAg2O·(100–x)TeO2 glass/SUS samples exhibit relatively low pH sensitivity. The pH sensitivities of the samples can be classified into two distinct groups. The first group, comprising samples with an Ag2O content of 25 mol% or more, exhibits a pH sensitivity higher than 10 %. The second group, comprising samples with an Ag2O content of 22 mol% or less, exhibits a pH sensitivity of less than 10 %. The pH sensitivity tends to decrease with increase in stutzite fraction in Ag–Te alloys. Results of X-ray diffraction measurements and transmission electron microscopy observations indicate that the hessite and stutzite Ag–Te alloys, which are minor and conductive, are the primary contributors to the low pH sensitivity observed. Moreover, the pH sensitivity of xAg2O·(100–x)TeO2/SUS is observed to be between those of the Ag–Te alloy and the Ag2O–TeO2 glass matrix. We propose occurrence of an electrical shunt should occur between the Ag–Te alloy and the Ag+-ion conductive Ag2O–TeO2 glass matrix. Regarding xAg2O·(100–x)TeO2 glass/SUS, the 20Ag2O·80TeO2 glass/SUS to 21Ag2O·79TeO2 glass/SUS compositions represent promising candidates as for KCl-leakage-free reference electrodes for pH sensors.

Abstract Background Prolonged laparoscopic nephroureterectomy (LNU) for upper tract urothelial cancer (UTUC) can increase the frequency of intravesical recurrence after surgery. Therefore, it is important for urological surgeons to have knowledge on preoperative risk factors for prolonged LNU. However, few studies have investigated the risk factors for prolonged LNU. We hypothesized that the quantity of perirenal fat affects the pneumoretroperitoneum time (PRT) of retroperitoneal LNU (rLNU). This study aimed to investigate the preoperative risk factors for prolonged PRT during rLNU. Methods We reviewed the data of 115 patients who underwent rLNU for UTUC between 2013 and 2021. The perirenal fat thickness (PFT) observed on preoperative computed tomography (CT) images was used to evaluate the perinephric fat quantity. Preoperative risk factors for PRT during rLNU were analyzed using logistic regression models. The cutoff value for PRT was determined based on the median time.The cutoff values for fat-related factors influencing PRT were defined according to receiver operating characteristic curve analysis. Results The median PRT for rLNU was 182 min (interquartile range, 155–230 min). The cutoff values of posterior, lateral, and anterior PFTs were 15 mm, 24 mm, and 6 mm, respectively. Multivariate analysis revealed that a posterior PFT ≥ 15 mm (odds ratio [OR], 2.72; 95% confidence interval, 1.04–7.08; p = 0.0410) was an independent risk factor for prolonged PRT. Conclusions Thick posterior PFT is a preoperative risk factor for prolonged PRT during rLNU. For patients with UTUC and thick posterior PFT, surgeons should develop optimal surgical strategies, including the selecting an expert surgeon as a primary surgeon and the selecting transperitoneal approach to surgery or open surgery.

Abstract Background The Vesical Imaging Reporting and Data System (VI-RADS) is widely used for predicting muscle-invasive bladder cancer (MIBC). This study aimed to determine the clinicopathological significance of the VI-RADS ≧4 (VI≧4) group. Methods Patients who underwent transurethral resections of bladder tumors during the study period and preoperative magnetic resonance imaging were considered. The patients were pathologically diagnosed with urothelial carcinoma (UC). We first compared the results of patients with VI-RADS scores of 3 and 4 to determine the cut-off score for MIBC; thereafter, the patients were divided into the VI≧4 and VI-RADS ≦3 (VI≦3) groups using VI-RADS. The clinicopathological significance of the VI≧4 group was examined retrospectively by comparing the characteristics of each group. Results In total, 121 cases were examined, of which 28 were pathologically diagnosed with MIBC. Of the 28 MIBC cases, three (10.7%) had a VI-RADS score of ≦3, and 25 (89.3%) had a VI-RADS score of ≧4. Of the 93 NMIBC cases, 86 (92.5%) had a VI-RADS score of ≦3, and seven (7.5%) had a VI-RADS score of ≧4. The diagnostic performance of the VI-RADS with a cut-off score of 4 was 89.3% for sensitivity, 92.5% for specificity, and an area under the curve (AUC) of 0.91. Contrastingly, for a cut-off score of 3, the sensitivity was 89.3%, specificity was 62.0%, and AUC was 0.72. A VI-RADS score of ≥ 4 could predict MIBC. In the VI≧4 group, 30 of 32 (93.8%) patients had high-grade tumors. The VI≧4 group had significantly more high-grade bladder cancers than the VI≦3 group (p

Abstract Background To investigate the impact of Perirenal fat stranding (PRFS) on progression after radical nephroureterectomy (RNU) for renal pelvic urothelial carcinoma (RPUC) without hydronephrosis and to reveal the pathological findings of PRFS. Methods Clinicopathological data, including computed tomography (CT) findings of the ipsilateral PRFS, were collected from the medical records of 56 patients treated with RNU for RPUC without hydronephrosis between 2011 and 2021 at our institution. PRFS on CT was classified as either low or high PRFS. The impact of PRFS on progression-free survival (PFS) after RNU was analyzed using the Kaplan–Meier method and log-rank test. In addition, specimens including sufficient perirenal fat from patients with low and with high PRFS were pathologically analyzed. Immunohistochemical analysis of CD68, CD163, CD3, and CD20 was also performed. Results Of the 56 patients, 31(55.4%) and 25 (44.6%) patients were classified as having low and high PRFS, respectively. Within a median follow-up of 40.6 months postoperatively, 11 (19.6%) patients showed disease progression. The Kaplan–Meier method and log-rank test revealed that patients with high PRFS had significantly lower PFS rates than those with low PRFS (3-year PFS 69.8% vs 93.3%; p = 0.0393). Pathological analysis revealed that high PRFS specimens (n = 3 patients) contained more fibrous strictures in perirenal fat than low PRFS specimens (n = 3 patients). In addition, M2 macrophages (CD163 +) infiltrating fibrous tissue in perirenal area were observed in all patients with high PRFS group. Conclusions PRFS of RPUC without hydronephrosis consists of collagenous fibers with M2 macrophages. The presence of ipsilateral high PRFS might be a preoperative risk factor for progression after RNU for RPUC patients without hydronephrosis. Prospective studies with large cohorts are required in the future.

Coronary artery lesions (CALs) after Kawasaki disease present complex coronary hemodynamics. We investigated the relationship between coronary fractional flow reserve (FFR), myocardial flow reserve (MFR), and myocardial blood flow volume fraction (MBF) and their clinical usefulness in CALs after Kawasaki disease. Nineteen patients (18 men, 1 woman) who underwent cardiac catheterization and 13N-ammonia positron emission tomography, with 24 coronary artery branches, were included. Five branches had inconsistent FFR and MFR values, two had normal FFR but abnormal MFR, and three had abnormal FFR and normal MFR. The abnormal MFR group had significantly higher MBF at rest than the normal group (0.86 ± 0.13 vs. 1.08 ± 0.09, p = 0.001). The abnormal FFR group had significantly lower MBF at adenosine loading than the normal group (2.23 ± 0.23 vs. 1.88 ± 0.29, p = 0.021). The three branches with abnormal FFR only had stenotic lesions, but the MFR may have been normal because blood was supplied by collateral vessels. Combining FFR, MFR, and MBF will enable a more accurate assessment of peripheral coronary circulation and stenotic lesions in CALs and help determine treatment strategy and timing of intervention.

Abstract Objective Clozapine may cause serious side effects despite benefits in patients with schizophrenia. Thus, an accurate understanding of the side-effect profile of clozapine is extremely important in the management of its administration to patients with schizophrenia. Our aim was to validate the relationship between clozapine exposure and appendicitis onset in patients with schizophrenia. Methods In this study, we retrospectively compared the incidence and cumulative incidence of appendicitis in patients with schizophrenia with and without a history of clozapine exposure. Among the patients with schizophrenia who visited our hospital between June 2009 and August 2021, we extracted those with a history of clozapine treatment. Patients with a history of taking clozapine were defined as the clozapine exposure group, while the others were defined as the clozapine non-exposure group. Patients with a history of appendectomy before their initial visit to our hospital or with a history of clozapine use at other hospitals were excluded. Results There were 65 patients in the clozapine exposure group and 400 patients in the clozapine non-exposure group who met the inclusion criteria. The exposure group exhibited a remarkably higher incidence of appendicitis during the observation period than the non-exposure group (863 cases vs. 124 cases per 100,000 person-years). In particular, if limited to the period of clozapine exposure, the incidence of appendicitis is extremely high, at 2,086 cases per 100,000 person-years. Moreover, multivariable analysis showed that clozapine exposure was an independent factor contributing to the onset of appendicitis. Conclusions Clozapine exposure is associated with appendicitis onset in patients with schizophrenia.

ABSTRACT. Although hospitalization for dementia is increasing, Japanese doctors often refrain from surgeries considering dementia. A woman in her 80s diagnosed with Alzheimer’s disease was admitted to hospital for cholelithiasis. Due to the avoidance of surgery, the inflammation was prolonged and therefore she was unable to eat. Later, she was discharged with central venous nutrition. The care burden on family resulted in her readmission to another hospital. Eventually, the inflammation was alleviated, and she was able to eat. However, it took a long time. In this study, we not only emphasize the risks but also focus on the benefits to postoperative rehabilitation. We also discuss about the benefits of invasive procedures in patients with dementia.

Abstract Background Active surveillance (AS) is one of the treatment methods for patients with small renal masses (SRMs;

A 56-year-old female patient with left breast cancer presented at our hospital. Preoperative CT scan showed an isolated bilateral pectoralis major muscle defect and abnormal muscle originating from the entire sternum and inserting in the lower ribs and rectus sheath. Total mastectomy and axillary lymph node dissection were performed. We believe that this case is unique and that others like it have never been reported. If there is a defect in the pectoralis major muscle, reconstructive surgery with a tissue expander is contraindicated. Therefore, preoperative evaluation of the chest wall musculature on imaging is recommended.

Metal nanoparticles (NPs), such as gold NPs (AuNPs), are particularly sensitive to X-rays, and thus specific accumulation of AuNPs in a tumor would allow radiotherapy with low energy X-rays and reduced side effects. AuNPs can be generated using HAuCl4 and the natural polyphenol epigallocatechin-3-gallate (EGCG) in the presence of citrate. Here, we generated EGCG-AuNPs in the presence of several additives and examined the accumulation of these NPs in mouse tumors following intravenous administration. EGCG-AuNPs 15 ​nm in diameter in the presence of sodium alginate accumulated more in tumors compared to 40-nm-diameter EGCG-AuNPs. Furthermore, the results of in vitro cellular uptake and serum protein absorption studies suggest that adsorption of 15–16 ​kDa serum proteins to EGCG-AuNPs suppresses accumulation in tumors. Thus, tendency to adsorb specific proteins on EGCG-AuNPs surface should be tailored for enhancing their accumulation in tumors.

Glass pH sensors are unsuitable for in vivo biomedical, clinical, or food applications because of the brittleness of glass and the difficulty in measuring small volumes. Enamel structures such as glass/stainless steel are candidates for glass-based pH electrodes. In this study, new enamel electrodes for pH sensors using Fe2O3–TeO2-based glass/stainless steel were developed. The effect of NiO addition to Fe2O3–TeO2 glass on the pH sensitivity and the three-point bending strength of enamels were investigated. The effect of NiO addition to Fe2O3–TeO2 glass/stainless steel on the pH sensitivity was negligible. Fe2O3–TeO2-based glass/stainless steel showed pH sensitivity appropriate to a working electrode. Enameling at a lower temperature under an air atmosphere was desirable for narrowing the gap between pH 4–7 and pH 7–9 sensitivities. The NiO addition to Fe2O3–TeO2 glass/stainless steel decreased the three-point bending strength. Therefore, NiO did not serve as an adhesion oxide in the Fe2O3–TeO2 glass. Fe2O3–TeO2 glass/stainless steel possessed the highest three-point bending strength among all samples when prepared at 670 °C under an air atmosphere. Therefore, no NiO addition and enameling at a lower temperature under an air atmosphere are desirable for obtaining more robust Fe2O3–TeO2 glass/stainless steel than Li2O–SiO2-based glass electrodes for pH sensors.

We report a case of spontaneous coronary artery dissection located next to a myocardial bridge in a patient with concomitant takotsubo cardiomyopathy. A fusion image with multidetector-row computed tomography and single-photon emission computed tomography played an important role in the diagnosis of these lesions. (Level of Difficulty: Advanced.)

The current work assessed the inclusion impacts of oilseed camelina meal (CM) as a protein source in red sea bream (Pagrus major) diets. A 45 day assessment period with 180 juveniles (6.47 ± 0.17 g) were allocated in triplicates to 4 experimental groups and fed formulated diets in which fish meal (FM) was subrogated at graded series of 0% (T1), 20.5% soybean meal (T2), 20.5% camelina meal (T3), and 33% camelina meal (T4). No noticeable alterations were observed in specific growth rate, feed intake, survival rate, hepatosomatic index, and Fulton's condition factor among the experimental groups. Fish fed the T4 diet showed considerably reduced (P 0.05) in glucose, total bilirubin, total protein, total cholesterol, triglyceride, glutamyl oxaloacetic transaminase, and glutamic pyruvate transaminase. Hematocrit levels decreased noticeably (P

Abstract Background Hair follicle (HF) formation and growth are sustained by epithelial-mesenchymal interaction via growth factors and cytokines. Pivotal roles of FGFs on HF regeneration and neogenesis have been reported mainly in rodent models. FGF expression is regulated by upstream pathways, represented by canonical WNT signaling; however, how FGFs influence on human folliculogenesis remains elusive. The aim of this study is to assess if human scalp-derived fibroblasts (sFBs) are able to modulate their FGF expression profile in response to WNT activation and to evaluate the influence of WNT-activated or suppressed FGFs on folliculogenesis. Methods Dermal papilla cells (DPCs), dermal sheath cells (DSCs), and sFBs were isolated from the human scalp and cultured independently. The gene expression profile of FGFs in DPCs, DSCs, and sFBs and the influence of WNT activator, CHIR99021, on FGF expression pattern in sFBs were evaluated by reverse transcription polymerase chain reaction, which were confirmed at protein level by western blotting analysis. The changes in the expression of DPC or keratinocyte (KC) biomarkers under the presence of FGF7 or 9 were examined in both single and co-culture assay of DPCs and/or KCs. The influence of FGF 7 and FGF 9 on hair morphogenesis and growth was analyzed in vivo using mouse chamber assay. Results In single culture, sFBs were distinguished from DPCs and DSCs by relatively high expression of FGF5 and FGF 18, potential inducers of hair cycle retardation or catagen phase. In WNT-activated state, sFBs downregulated FGF7 while upregulating FGF9, a positive regulator of HF morphogenesis, FGF16 and FGF 20 belonging to the same FGF subfamily. In addition, CHIR99021, a WNT activator, dose-dependently modulated FGF7 and 9 expression to be folliculogenic. Altered expressions of FGF7 and FGF9 by CHIR99021 were confirmed at protein level. Supplementation of FGF9 to cultured DPCs resulted in upregulation of representative DP biomarkers and this tendency was sustained, when DPCs were co-cultured with KCs. In mouse chamber assay, FGF9 increased both the number and the diameter of newly formed HFs, while FGF7 decreased HF diameter. Conclusion The results implied that sFBs support HF formation by modulating regional FGF expression profile responding to WNT activation.

Abstract Background Statins suppress the progression of atherosclerosis by reducing low-density lipoprotein (LDL) cholesterol levels. Pemafibrate (K-877), a novel selective peroxisome proliferator-activated receptor α modulator, is expected to reduce residual risk factors including high triglycerides (TGs) and low high-density lipoprotein (HDL) cholesterol during statin treatment. However, it is not known if statin therapy with add-on pemafibrate improves the progression of atherosclerosis. The aim of this study was to assess the effect of combination therapy with pitavastatin and pemafibrate on lipid profiles and endothelial dysfunction in hypertension and insulin resistance model rats. Methods Seven-week-old male Dahl salt-sensitive (DS) rats were divided into the following five treatment groups (normal diet (ND) plus vehicle, high-salt and high-fat diet (HD) plus vehicle, HD plus pitavastatin (0.3 mg/kg/day), HD plus pemafibrate (K-877) (0.5 mg/kg/day), and HD plus combination of pitavastatin and pemafibrate) and treated for 12 weeks. At 19 weeks, endothelium-dependent relaxation of the thoracic aorta in response to acetylcholine was evaluated. Results After feeding for 12 weeks, systolic blood pressure and plasma levels of total cholesterol were significantly higher in the HD-vehicle group compared with the ND-vehicle group. Combination therapy with pitavastatin and pemafibrate significantly reduced systolic blood pressure, TG levels, including total, chylomicron (CM), very LDL (VLDL), HDL-TG, and cholesterol levels, including total, CM, VLDL, and LDL-cholesterol, compared with vehicle treatment. Acetylcholine caused concentration-dependent relaxation of thoracic aorta rings that were pre-contracted with phenylephrine in all rats. Relaxation rates in the HD-vehicle group were significantly lower compared with the ND-vehicle group. Relaxation rates in the HD-combination of pitavastatin and pemafibrate group significantly increased compared with the HD-vehicle group, although neither medication alone ameliorated relaxation rates significantly. Western blotting experiments showed increased phosphorylated endothelial nitric oxide synthase protein expression in aortas from rats in the HD-pemafibrate group and the HD-combination group compared with the HD-vehicle group. However, the expression levels did not respond significantly to pitavastatin alone. Conclusions Combination therapy with pitavastatin and pemafibrate improved lipid profiles and endothelial dysfunction in hypertension and insulin resistance model rats. Pemafibrate as an add-on strategy to statins may be useful for preventing atherosclerosis progression.

Abstract Background The rabbit squamous cell cancer line, VX2, has been used to generate various tumor models in rabbits. It is notable for its ability to generate nodal metastases. However, the timing and extent of nodal metastases vary by primary inoculation site and methodology. The development of metastases specifically in lung cancer models has not been well-described. We sought to characterize the generation of nodal metastases in rabbit transbronchial VX2 lung tumor models. Methods Rabbit VX2 lung tumor models were created in the right lung via transbronchial injection and serially imaged by computed tomography. Rabbits (n = 15) were sacrificed from between 5 and 24 days post-inoculation for collection of the ipsilateral and contralateral paratracheal lymph nodes. These underwent histopathological evaluation for metastases using hematoxylin and eosin as well as cytokeratin AE1/AE3 immunohistochemical staining. Results Nodal metastases were detectable as early as 1 week after inoculation but were more prevalent with longer inoculation; all rabbits at > 2 weeks post-inoculation had nodal metastases. Contralateral metastases were in general seen later than ipsilateral metastases. Lymph node volume did not predict the likelihood of nodal metastases (p = 0.4 and p = 0.07 for ipsilateral and contralateral nodal metastases, respectively), but primary tumor volume was significantly associated with the likelihood of nodal metastases (p = 0.001 and p = 0.005 for ipsilateral and contralateral nodal metastases, respectively). Ipsilateral metastases were detectable at a tumor diameter of 1 cm; contralateral metastases were more variable but in general required a tumor diameter of 2 cm. Conclusions Rabbit transbronchial VX2 lung tumor models generate nodal metastases relatively early after inoculation. These results suggest such models may be valuable tools in the investigation of novel therapeutic modalities relevant for the treatment of both early-stage and locally advanced lung cancer.

Abstract Background Novel feeding adaptations often facilitate adaptive radiation and diversification. But the evolutionary origins of such feeding adaptations can be puzzling if they require concordant change in multiple component parts. Pelagic, heterorhabdid copepods (Calanoida) exhibit diverse feeding behaviors that range from simple particle feeding to a highly specialized form of carnivory involving piercing mouthparts that likely inject venom. We review the evolutionary history of heterorhabdid copepods and add new high-resolution, 3D anatomical analyses of the muscular system, glands and gland openings associated with this remarkable evolutionary transformation. Results We examined four heterorhabdid copepods with different feeding modes: one primitive particle-feeder (Disseta palumbii), one derived and specialized carnivore (Heterorhabdus subspinifrons), and two intermediate taxa (Mesorhabdus gracilis and Heterostylites longicornis). We used two advanced, high-resolution microscopic techniques — serial block-face scanning electron microscopy and two-photon excitation microscopy — to visualize mouthpart form and internal anatomy at unprecedented nanometer resolution. Interactive 3D graphical visualizations allowed putative homologues of muscles and gland cells to be identified with confidence and traced across the evolutionary transformation from particle feeding to piercing carnivory. Notable changes included: a) addition of new gland cells, b) enlargement of some (venom producing?) glands, c) repositioning of gland openings associated with hollow piercing fangs on the mandibles, d) repurposing of some mandibular-muscle function to include gland-squeezing, and e) addition of new muscles that may aid venom injection exclusively in the most specialized piercing species. In addition, live video recording of all four species revealed mandibular blade movements coupled to cyclic contraction of some muscles connected to the esophagus. These behavioral and 3D morphological observations revealed a novel injection system in H. subspinifrons associated with piercing (envenomating?) carnivory. Conclusions Collectively, these results suggest that subtle changes in mandibular tooth form, and muscle and gland form and location, facilitated the evolution of a novel, piercing mode of feeding that accelerated diversification of the genus Heterorhabdus. They also highlight the value of interactive 3D animations for understanding evolutionary transformations of complex, multicomponent morphological systems.

The euryhaline copepod Pseudodiaptomus inopinus play important roles in coastal waters as vectors of docosahexanoic (DHA) and eicosapentaenoic (EPA) acids for larval fish. While DHA and EPA in polar lipids (PLs) are more effective for fish larval development than non-polar lipid forms (NLs), there is little knowledge how much these lipids are accumulated in copepods from microalgae and are effective for early development of fish larvae. We report PLs fatty acid profiles of P. inopinus fed DHA-poor microalgae and evaluate its significance as a food source for larvae development of Pagrus major, compared with DHA-enriched rotifers. Copepods and rotifers were fed a mixed diet of three algal species (Phaeodactylum tricornutum, Tisochrysis lutea, and Pavlova lutheri), in addition of DHA-supplemented Super Fresh Chlorella (SFC) for rotifers. Compared with SFC, the algal mixture had higher EPA but lower DHA. Copepods had higher DHA and EPA in total lipids than rotifers fed each diet. Copepod PLs were specifically enriched with DHA and their contents were higher than both rotifers. On the other hand, PLs EPA contents were comparable between preys, indicating that copepods selectively fortified the PLs. Fish culture experiment showed that larvae fed copepods had higher growth than those fed SFC-enriched rotifers. Principal component analysis for each organism fatty acid composition emphasized trophic modification of DHA by copepods toward larval fish. This study highlighted that P. inopinus contribute to enhanced growth of coastal larval fish by efficiently transferring DHA via copepod fatty acid metabolism.

Background Sinus tachycardia during exercise attenuates ST‐segment elevation in patients with Brugada syndrome, whereas ST‐segment augmentation after an exercise test is a high‐risk sign. Some patients have premature ventricular contractions (PVCs) related to exercise, but the significance of exercise‐related PVCs in patients with Brugada syndrome is still unknown. The objective of this study was to determine the significance of exercise‐related PVCs for predicting occurrence of ventricular fibrillation (VF) in patients with Brugada syndrome. Methods and Results The subjects were 307 patients with Brugada syndrome who performed a treadmill exercise test. We evaluated the occurrence of PVCs at rest, during exercise and at the peak of exercise, and during recovery after exercise (0–5 minutes). We followed the patients for 92±68 months and evaluated the occurrence of VF. PVCs occurred in 82 patients (27%) at the time of treadmill exercise test: PVCs appeared at rest in 14 patients (4%), during exercise in 60 patients (20%), immediately after exercise (0–1.5 minutes) in 28 patients (9%), early after exercise (1.5–3 minutes) in 18 patients (6%), and late after exercise (3–5 minutes) in 12 patients (4%). Thirty patients experienced VF during follow‐up. Multivariable analysis including symptoms, spontaneous type 1 ECG, and PVCs in the early recovery phase showed that these factors were independently associated with VF events during follow‐up. Conclusions PVCs early after an exercise test are associated with future occurrence of VF events. Rebound of vagal nerve activity at the early recovery phase would promote ST‐segment augmentation and PVCs in high‐risk patients with Brugada syndrome.

Deep learning is being increasingly applied for obtaining digital microscopy image data of cells. Well-defined annotated cell images have contributed to the development of the technology. Cell morphology is an inherent characteristic of each cell type. Moreover, the morphology of a cell changes during its lifetime because of cellular activity. Artificial intelligence (AI) capable of recognizing a mouse-induced pluripotent stem (miPS) cell cultured in a medium containing Lewis lung cancer (LLC) cell culture-conditioned medium (cm), miPS-LLCcm cell, which is a cancer stem cell (CSC) derived from miPS cell, would be suitable for basic and applied science. This study aims to clarify the limitation of AI models constructed using different datasets and the versatility improvement of AI models. The trained AI was used to segment CSC in phase-contrast images using conditional generative adversarial networks (CGAN). The dataset included blank cell images that were used for training the AI but they did not affect the quality of predicting CSC in phase contrast images compared with the dataset without the blank cell images. AI models trained using images of 1-day culture could predict CSC in images of 2-day culture; however, the quality of the CSC prediction was reduced. Convolutional neural network (CNN) classification indicated that miPS-LLCcm cell image classification was done based on cultivation day. By using a dataset that included images of each cell culture day, the prediction of CSC remains to be improved. This is useful because cells do not change the characteristics of stem cells owing to stem cell marker expression, even if the cell morphology changes during culture.

Previously, we have succeeded in converting induced pluripotent stem cells (iPSCs) into cancer stem cells (CSCs) by treating the iPSCs with conditioned medium of Lewis lung carcinoma (LLC) cells. The converted CSCs, named miPS-LLCcm cells, exhibited the self-renewal, differentiation potential, and potential to form malignant tumors with metastasis. In this study, we further characterized miPS-LLCcm cells both in vivo and in vitro. The tumors formed by subcutaneous injection showed the structures with pathophysiological features consisting of undifferentiated and malignant phenotypes generally found in adenocarcinoma. Metastasis in the lung was also observed as nodule structures. Excising from the tumors, primary cultured cells from the tumor and the nodule showed self-renewal, differentiation potential as well as tumor forming ability, which are the essential characters of CSCs. We then characterized the epigenetic regulation occurring in the CSCs. By comparing the DNA methylation level of CG rich regions, the differentially methylated regions (DMRs) were evaluated in all stages of CSCs when compared with the parental iPSCs. In DMRs, hypomethylation was found superior to hypermethylation in the miPS-LLCcm cells and its derivatives. The hypo- and hypermethylated genes were used to nominate KEGG pathways related with CSC. As a result, several categories were defined in the KEGG pathways from which most related with cancers, significant and high expression of components was PI3K-AKT signaling pathway. Simultaneously, the AKT activation was also confirmed in the CSCs. The PI3K-Akt signaling pathway should be an important pathway for the CSCs established by the treatment with conditioned medium of LLC cells.

Abstract Background Aphids display “cyclic parthenogenesis,” in which parthenogenetically and sexually reproducing morphs seasonally alternate in the aphid annual life cycles. There are various characteristics that differ between asexual viviparous and sexual oviparous females. In oviparous females, swollen cuticular structures (~ 10 μm in diameter), called “scent plaques,” are scattered on the surface of hind tibias, and secrete monoterpenoid sex pheromones. However, the developmental processes of the pheromone glands and the biosynthetic pathways of monoterpenoid pheromones have yet to be elucidated. Results Comparisons of the developmental processes that form hind tibias between sexual and parthenogenetic females revealed that, in sexual females, the epithelial tissues in proximal parts of hind tibias become columnar in fourth instar nymphs, and circular pheromone glands with Class 1 gland cells appear in adults, although they do not appear in parthenogenetic females. Furthermore, by comparing the expression levels of genes involved in the mevalonate pathway, which is required for monoterpenoid synthesis, we show that genes that encode the downstream enzymes in the pathway are highly expressed in hind tibias of sexual females. Conclusion Glandular tissues of scent plaque are differentiated from the fourth instar in sexual females, while parthenogenetic females lack the glandular cells. Only the downstream steps of the mevalonate pathway appear to occur in scent plaques on hind tibias of sexual females, although the upstream steps may occur somewhere in other body parts.

Lipidomics, the mass spectrometry-based comprehensive analysis of lipids, has attracted attention as an analytical approach to provide novel insight into lipid metabolism and to search for biomarkers. However, an ideal method for both comprehensive and quantitative analysis of lipids has not been fully developed. Here, we have proposed a practical methodology for widely targeted quantitative lipidome analysis using supercritical fluid chromatography fast-scanning triple-quadrupole mass spectrometry (SFC/QqQMS) and theoretically calculated a comprehensive lipid multiple reaction monitoring (MRM) library. Lipid classes can be separated by SFC with a normal-phase diethylamine-bonded silica column with high resolution, high throughput, and good repeatability. Structural isomers of phospholipids can be monitored by mass spectrometric separation with fatty acyl-based MRM transitions. SFC/QqQMS analysis with an internal standard-dilution method offers quantitative information for both lipid class and individual lipid molecular species in the same lipid class. Additionally, data acquired using this method has advantages, including reduction of misidentification and acceleration of data analysis. Using the SFC/QqQMS system, alteration of plasma lipid levels in myocardial infarction-prone rabbits to the supplementation of EPA was first observed. Our developed SFC/QqQMS method represents a potentially useful tool for in-depth studies focused on complex lipid metabolism and biomarker discovery.—Takeda, H., Y. Izumi, M. Takahashi, T. Paxton, S. Tamura, T. Koike, Y. Yu, N. Kato, K. Nagase, M. Shiomi, and T. Bamba.

A 60-day experiment was designed to assess the effect of different ratios of fish meal (FM): camelina meal plant protein (CM) on growth response and relative gene expression of growth-promoting factors, feed utilization potency, digestive enzymes activities, apparent digestibility (ADC), stress response, non-specific immunity of Pagrus major. Four isonitrogenous (490.7 g/kg of crude protein) and isolipidic (91.5 g/kg total lipid) experimental diets were formulated and designated as camelina meal (CM0), soyabean meal (SBM20.5), CM20.5, and CM33 based on protein contents. At the end of the feed trial, significantly higher (p < 0.05) weight gain, specific growth rate, and feed intake but lower feed conversion ratio were recorded in fish fed CM0, SBM20.5, and CM20.5 than fish fed CM33. The lowest growth, feed utilization, enzyme activity, and digestibility were recorded in fish fed CM33. Significantly higher pepsin, amylase, and protease activities were observed in fish fed CM0, SBM20.5, and CM20.5 diets than fish fed CM33. The highest ADC of protein was recorded in fish fed CM0, SBM20.5, and CM20.5 diets. Hematocrit levels were depressed CM33 while total serum protein, total cholesterol, triglyceride, blood urea nitrogen, total bilirubin, aspartate aminotransferase, and alanine aminotransferase were not significantly changed by the inclusion of CM. Non-specific immune variables (lysozyme activity, peroxidase activity in serum and nitro blue tetrazolium) in fish fed CM0, SBM20.5, and CM20.5 were significantly higher than in fish fed CM33 diet. The superoxide dismutase of fish fed CM20.5 was not significantly different from CM0 and SBM20.5 (p > 0.05). Catalase and low salinity stress test show that CM0, SBM20.5, and CM20.5 were not significantly (p > 0.05) different, while CM33 was significantly lower than the rest of the diets. TBARs show that CM20.5 and CM33 diets were significantly different (p < 0.05), but CM20.5 was not significantly different from SBM20.5. Significantly higher hepatic IGF-1 and IGF-2 mRNA expression was found in fish-fed diet groups CM0, SBM20.5, and CM20.5 than fish fed CM33. The present study indicated that the addition of CM up 205 kg/kg to diet maintains growth, digestive enzymes, nutrient digestibility, immunity, stress resistance, and feed utilization efficiency of red sea bream.

The three major conventional treatments: surgery, chemotherapy, and radiation therapy, have been commonly performed for lung cancer. However, lung cancer is still the leading cause of cancer-related mortality. Immunotherapy has recently emerged as a very effective new treatment modality, and there is now growing enthusiasm for cancer immunotherapy worldwide. However, the results of clinical studies using immunotherapy are not always favorable. Understanding the steps involved in the recognition and eradication of cancer cells by the immune system seems essential to understanding why past immunotherapies have failed and how current therapies can be optimally utilized. In addition, the combination of immunotherapies, such as cancer vaccines and immune checkpoint inhibitors, as well as the combination of these therapies with three conventional therapies, may pave the way for personalized immunotherapy. In this review, we summarize the results of immunotherapies used in phase III clinical trials, including immune checkpoint inhibitors, and discuss the future prospects of immunotherapies in lung cancer treatment.

Abstract Cancer-associated fibroblasts (CAFs) are one of the most prominent cell types in the stromal compartment of the tumor microenvironment. CAFs support multiple aspects of cancer progression, including tumor initiation, invasion, and metastasis. The heterogeneous nature of the stromal microenvironment is attributed to the multiple sources from which the cells in this compartment originate. The present study provides the first evidence that cancer stem cells (CSCs) are one of the key sources of CAFs in the tumor niche. We generated CSC-like cells by treating mouse induced pluripotent stem cells with conditioned medium from breast cancer cell lines. The resulting cell population expressed both CSC and pluripotency markers, and the sphere-forming CSC-like cells formed subcutaneous tumors in nude mice. Intriguingly, these CSC-like cells always formed heterogeneous populations surrounded by myofibroblast-like cells. Based on this observation, we hypothesized that CSCs could be the source of the CAFs that support tumor maintenance and survival. To address this hypothesis, we induced the differentiation of spheres and purified the myofibroblast-like cells. The resulting cells exhibited a CAF-like phenotype, suggesting that they had differentiated into the subpopulations of cells that support CSC self-renewal. These findings provide novel insights into the dynamic interplay between various microenvironmental factors and CAFs in the CSC niche.

An amendment to this paper has been published and can be accessed via the original article.

PurposeDiagnosis and resection of indeterminate pulmonary nodules (IPNs) is a growing challenge with increased utilization of chest computed tomography. Photoacoustic (PA) -guided surgical resection with local injection of indocyanine green (ICG) may have utility for IPNs that are suspicious for lung cancer. This preclinical study explores the potential of PA imaging (PAI) to detect ICG-labeled tumors.Materials and methodsICG uptake by H460 lung cancer cells was evaluated in vitro. A phantom study was performed to analyze PA signal intensity according to ICG concentration and tissue thickness/depth using chicken breast. PA signals were measured up to 48 hours after injection of ICG (mixed with 5% agar) into healthy subcutaneous tissue, subcutaneous H460 tumors and right healthy lung in nude mice.ResultsIntracellular ICG fluorescence was detected in H460 cells co-incubated with ICG in vitro. The concentration dependence of the PA signal was logarithmic, and PA signal decline was exponential with increasing tissue depth. The PA signal of 2 mg/mL ICG was still detectable at a depth of 22 mm in chicken breast. The PA signal from ICG mixed with agar was detectable 48 hours post injection into subcutaneous tissue and subcutaneous H460 tumors in nude mice. Similar features of PA signals from ICG-agar in mice lung were obtained.ConclusionThe results from this preclinical study suggests that PAI of injected ICG-agar may be beneficial for identifying deeply located tumors. These features may be valuable for IPNs.

Investigation of magnetic properties and nanostructure of stacked media with a CoPt-B2O3 granular recording layer (GRL) and granular cap layers (CL) with CoPt and a typical ferromagnetic oxide of rare earth (RE) oxide (RE oxide: Gd2O3, Nd2O3, Sm2O3, and CeO2) are reported. CoPt-RE oxide granular layer shows higher saturation magnetization and magnetic anisotropy than that of CoPtCrB continuous layer. In stacked media with CoPt-Gd2O3 granular CL (9 nm), nucleation field increases from 0.7 to 0.8 kOe and saturation field decreases from 21.5 to 12.5 kOe with comparable values to those of CoPtCrB continuous CL. This indicates that CoPt-RE oxide granular CL is effective to control intergranular exchange coupling. Reduction of coercivity distribution ratio (∆Hc/Hc) from 0.15 to 0.09 which is comparable to that of CoPtCrB continuous CL when CoPt-Gd2O3 granular CL thickness is changed from 4 to 9 nm is observed. This means CoPt-Gd2O3 granular CL is effective to reduce switching field distribution. Furthermore, significant reduction of surface roughness (Ra) for media with CoPt-Gd2O3 granular CL (9 nm) from 0.32 to 0.25 nm is observed. This reveals that CoPt-RE oxide granular CL has a great impact to induce stacked media with flat surface. Based on the cross-section observation by TEM, the media with CoPt-Gd2O3 granular CL shows continuous structure of grain boundaries from the GRL up to the CL and which results in one-on-one growth of magnetic grain with homogeneous columnar structure. Focusing on the surface morphology, media with CoPt-Gd2O3 granular CL shows smoother surface than that of media with CoPtCrB continuous CL which supports the Ra evaluation result. These results indicate that the employment of granular CoPt-RE oxide for CL is not only to reduce switching field distribution but also to flatten surface of the stacked media.

Gastrointestinal stromal tumors (GISTs) of the stomach presenting as an intra-abdominal abscess are extremely rare. We herein report a case that underwent successful laparoscopic resection of gastric GIST presenting with an intra-abdominal abscess. A 70-year-old man presented with a 3-day history of acute upper abdominal pain with a fever. Laboratory data showed an elevated white blood cell count and C-reactive protein level. A CT scan revealed a mass of mixed solid and fluid components, measuring 5.5 cm, in the adjacent lesser curvature of the stomach. He was diagnosed as having an intra-abdominal abscess, and antibiotic therapy was administered. Consequently, his symptoms were relieved and a CT scan on the 7th day showed that the mass reduced and became homogenous. According to the results of a fine-needle aspiration biopsy, the tumor was suspected to be a GIST. Laparoscopic surgery with the 5-port approach was performed. The tumor existed in the lesser curvature of the stomach, and was excised en bloc with the omentum and gastric wall by wedge resection. The tumor measured 3.0 cm in diameter and originated from the serosal layer of the stomach. Histological findings showed spindle cells with a stromal growth pattern that was positive for c-kit (CD 117) and CD 34. The postoperative course was uneventful, and no recurrence has been noted in the 2 years since the operation. This is the first report of a case with gastric GIST with an intra-abdominal abscess that was successfully treated by laparoscopic surgery.

Hyperlipidemia or dyslipidemia is a major risk factor for atherosclerotic diseases. Experimental animals play an important role in elucidating the molecular mechanisms of the pathophysiology of hyperlipidemia as well as in drug development. Rabbits are one of the most suitable models to study human hyperlipidemia because many features of the lipoprotein metabolism of rabbits are similar to those of humans such as LDL-rich lipoproteins in plasma, apolipoprotein B mRNA editing, and cholesteryl ester transfer protein. Currently, three types of rabbit models are commonly used for studying hyperlipidemia: (1) diet-induced hyperlipidemic rabbits, (2) spontaneous hyperlipidemic rabbits, and (3) gene-manipulated rabbits (transgenic and knockout rabbits). In this review, we give an overview of the features of hyperlipidemic rabbits and discuss the usefulness of rabbits for the development of anti-atherogenic drugs.

Deep-learning workflows of microscopic image analysis are sufficient for handling the contextual variations because they employ biological samples and have numerous tasks. The use of well-defined annotated images is important for the workflow. Cancer stem cells (CSCs) are identified by specific cell markers. These CSCs were extensively characterized by the stem cell (SC)-like gene expression and proliferation mechanisms for the development of tumors. In contrast, the morphological characterization remains elusive. This study aims to investigate the segmentation of CSCs in phase contrast imaging using conditional generative adversarial networks (CGAN). Artificial intelligence (AI) was trained using fluorescence images of the Nanog-Green fluorescence protein, the expression of which was maintained in CSCs, and the phase contrast images. The AI model segmented the CSC region in the phase contrast image of the CSC cultures and tumor model. By selecting images for training, several values for measuring segmentation quality increased. Moreover, nucleus fluorescence overlaid-phase contrast was effective for increasing the values. We show the possibility of mapping CSC morphology to the condition of undifferentiation using deep-learning CGAN workflows.

Investigation of magnetic properties and microstructure of granular media with various multiple oxides as the grain boundary material is reported. Saturation magnetization (Ms), uniaxial magnetocrystalline anisotropy (Ku), and magnetic grain diameter (GD) of the granular media show linear correlation with volume weighted average for melting point (Tm) of each oxides (Tmave). Ku of magnetic grains (Kugrain) shows a trade-off relation with GD that it is a big challenge to satisfy both high Kugrain and small GD by only controlling Tmave. To obtain a granular medium with appropriate Kugrain, GD, and low degree of intergranular exchange coupling, the combination of Tmave control of grain boundary material by mixing oxides and employment of a buffer layer are required. Here the degree of intergranular exchange coupling is estimated from the slope of M-H loop at around coercivity (α). By applying this technique, a typical granular medium with Kugrain of 1.0×107 erg/cm3, GD of 5.1 nm, and α of 1.2 is realized.

We performed gene expression microarray analysis coupled with spherical self-organizing map (sSOM) for artificially developed cancer stem cells (CSCs). The CSCs were developed from human induced pluripotent stem cells (hiPSCs) with the conditioned media of cancer cell lines, whereas the CSCs were induced from primary cell culture of human cancer tissues with defined factors ( OCT3/4, SOX2 , and KLF4 ). These cells commonly expressed human embryonic stem cell (hESC)/hiPSC-specific genes ( POU5F1, SOX2, NANOG, LIN28 , and SALL4 ) at a level equivalent to those of control hiPSC 201B7. The sSOM with unsupervised method demonstrated that the CSCs could be divided into three groups based on their culture conditions and original cancer tissues. Furthermore, with supervised method, sSOM nominated TMED9, RNASE1, NGFR, ST3GAL1, TNS4, BTG2, SLC16A3, CD177, CES1, GDF15, STMN2, FAM20A, NPPB, CD99, MYL7, PRSS23, AHNAK , and LOC152573 genes commonly upregulating among the CSCs compared to hiPSC, suggesting the gene signature of the CSCs.

This study reports on a 70-year-old man with recurrent cervical myelopathy 20 years after anterior decompression and fusion of C4–7 using a free vascularised strut graft. The recurrent myelopathy was secondary to a kyphotic deformity of a fractured graft and residual ossification of the posterior longitudinal ligament with stenosis at C3/4. Intra-operative spinal cord–evoked potentials indicated that spinal cord traction secondary to progressive kyphosis of the cervical spine after the graft fracture was the cause. The patient underwent laminoplasty at C3 and laminectomy at C4 to decompress the stenosis at C3/4 as well as posterior cervical spinal fusion at C3–7 with pedicle screws and a lateral mass screw and a bone graft to prevent further progression of the kyphosis. At postoperative 18 months, the patient's Japanese Orthopaedic Association score had improved to 14 from 8, and he could walk without support.

Taxanes including paclitaxel and docetaxel are effective anticancer agents preferably sufficient for liposomal drug delivery. However, the encapsulation of these drugs with effective amounts into conventional liposomes is difficult due to their high hydrophobicity. Therefore, an effective encapsulation strategy for liposomal taxanes has been eagerly anticipated. In this study, the mixture of polyethoxylated castor oil (Cremophor EL) and ethanol containing phosphate buffered saline termed as CEP was employed as a solvent of the inner hydrophilic core of liposomes where taxanes should be incorporated. Docetaxel-, paclitaxel-, or 7-oxacetylglycosylated paclitaxel-encapsulating liposomes were successfully prepared with almost 100% of encapsulation efficiency and 29.9, 15.4, or 29.1 mol% of loading efficiency, respectively. We then applied the docetaxel-encapsulating liposomes for targeted drug delivery. Docetaxel-encapsulating liposomes were successfully developed HER2-targeted drug delivery by coupling HER2-specific binding peptide on liposome surface. The HER2-targeting liposomes exhibited HER2-specific internalization and enhanced anticancer activity in vitro. Therefore, we propose the sophisticated preparation of liposomal taxanes using CEP as a promising formulation for effective cancer therapies.

Although the encapsulation of paclitaxel into liposomes has been extensively studied, its significant hydrophobic and uncharged character has generated substantial difficulties concerning its efficient encapsulation into the inner water core of liposomes. We found that a more hydrophilic paclitaxel molecule, 7-glucosyloxyacetylpaclitaxel, retained tubulin polymerization stabilization activity. The hydrophilic nature of 7-glucosyloxyacetylpaclitaxel allowed its efficient encapsulation into the inner water core of liposomes, which was successfully accomplished using a remote loading method with a solubility gradient between 40% ethylene glycol and Cremophor EL/ethanol in PBS. Trastuzumab was then conjugated onto the surface of liposomes as immunoliposomes to selectively target human epidermal growth factor receptor-2 (HER2)-overexpressing cancer cells. In vitro cytotoxicity assays revealed that the immunoliposomes enhanced the toxicity of 7-glucosyloxyacetylpaclitaxel in HER2-overexpressing cancer cells and showed more rapid suppression of cell growth. The immunoliposomes strongly inhibited the tumor growth of HT-29 cells xenografted in nude mice. Notably, mice survived when treated with the immunoliposomes formulation, even when administered at a lethal dose of 7-glucosyloxyacetylpaclitaxel in vivo. This data successfully demonstrates immunoliposomes as a promising candidate for the efficient delivery of paclitaxel glycoside.

Cancer stem cells (CSCs) are capable of continuous proliferation and self-renewal and are proposed to play significant roles in oncogenesis, tumor growth, metastasis and cancer recurrence. CSCs are considered derived from normal stem cells affected by the tumor microenvironment although the mechanism of development is not clear yet. In 2007, Yamanaka's group succeeded in generating Nanog mouse induced pluripotent stem (miPS) cells, in which green fluorescent protein (GFP) has been inserted into the 5'-untranslated region of the Nanog gene. Usually, iPS cells, just like embryonic stem cells, are considered to be induced into progenitor cells, which differentiate into various normal phenotypes depending on the normal niche. We hypothesized that CSCs could be derived from Nanog miPS cells in the conditioned culture medium of cancer cell lines, which is a mimic of carcinoma microenvironment. As a result, the Nanog miPS cells treated with the conditioned medium of mouse Lewis lung carcinoma acquired characteristics of CSCs, in that they formed spheroids expressing GFP in suspension culture, and had a high tumorigenicity in Balb/c nude mice exhibiting angiogenesis in vivo. In addition, these iPS-derived CSCs had a capacity of self-renewal and expressed the marker genes, Nanog, Rex1, Eras, Esg1 and Cripto, associated with stem cell properties and an undifferentiated state. Thus we concluded that a model of CSCs was originally developed from miPS cells and proposed the conditioned culture medium of cancer cell lines might perform as niche for producing CSCs. The model of CSCs and the procedure of their establishment will help study the genetic alterations and the secreted factors in the tumor microenvironment which convert miPS cells to CSCs. Furthermore, the identification of potentially bona fide markers of CSCs, which will help the development of novel anti-cancer therapies, might be possible though the CSC model.