Liquid biopsy with circulating tumor DNA (ctDNA) has become widely applied in clinical settings in step with progress in innovative technologies. Since circulating free DNA (cfDNA) was found in human plasma from patients with various cancers as well as auto-immune diseases, clinical application to oncology has spread, especially involving molecular analysis. cfDNA contains ctDNA derived from cancer cells, and it is not possible to isolate ctDNA from cfDNA derived from normal cells. Therefore, we need to recognize ctDNA by detection of somatic mutations corresponding to those observed in cancer tissue. Early on, these mutations were detected by polymerase chain reaction-based methods such as digital PCR. Recently, comprehensive genomic analysis with next generation sequencing (NGS) has been developed for liquid biopsy. Clinical applications include monitoring of tumor progression, analysis of mechanisms of resistance to molecular targeted therapy, and the possibility of detecting minimum residual disease after surgery. However, the sensitivity of ctDNA detection is unsatisfactory, and application is still limited to advanced cancers. To solve these problems, it is necessary to perform basic analysis of ctDNA and establish an efficient system for ctDNA isolation. Recent investigations showed that the cfDNA size distribution differs between ctDNA and cfDNA derived from normal cells, leading to efficient detection of ctDNA. Such new concepts could lead to greater development of liquid biopsy.

Immune resilience is the ability for the human body to adapt and respond to adverse conditions, such as bacterial pathogens, viral infections, and abnormal or cancerous cells. Our ability to mount an appropriate response against such illness relates directly to our level of immune resilience, which in turn is impacted by our lifestyle choices. Nutrition, exercise, sleep and stress management are powerful influencers of mucosal immunity, the first line of defense against pathogens, and the exposome, exogenous and endogenous factors affecting our overall health. Lifestyle medicine offers evidence-based recommendations on how to optimize our immune system to withstand and thwart infections and disease.

Purpose: Helical tomotherapy (HT) is one of the radiotherapy methods that has become widespread in recent years. The aim of this study is to investigate the safety of high-dose HT for medically inoperable esophageal cancer that is unable to tolerate chemoradiotherapy.

Methods: Eight patients with medically inoperable esophageal squamous cell carcinoma who cannot tolerate chemoradiotherapy were irradiated to 60 Gy at 2 Gy/fraction with HT.

Results: The 1-, and 2-year overall survival rate were 75.0%, and 46.9%, respectively. The 1-, and 2-year locoregional recurrence-free survival rate ware 72.9%, and 72.9%, respectively. Three patients developed grade 2 or 3 hematological acute toxicity, and one patient developed grade 2 late pericardial effusion.

Conclusions: Medically inoperable patients with esophageal cancer who cannot tolerate chemoradiotherapy might be treated safely by high-dose HT with minimal adverse events.

Purpose: Two anti-TNF biologics, infliximab and etanercept, are extremely useful for inflammatory diseases such as rheumatoid arthritis (RA). However, approximately 20 to 30% of RA have been described as non-responders. Here, we analyzed the statistical relationships of whole genome single nucleotide polymorphisms (SNPs) with remission or low disease activity (LDA) among RA and developed algorithms using SNPs to predict effectiveness of these biologics.

Methods: The total study subjects (first, second and validation population for each infliximab and etanercept) were 260 RA patients for infliximab and another 251 RA patients for etanercept. Effectiveness of infliximab and etanercept was assessed using DAS28-CRP. In first, second and the combined population, relationships of 277,339 SNPs with remission and LDA were analyzed using case-control analyses by Fisher's exact tests for each infliximab and etanercept. We picked up SNPs (P < 0.05) from each first, second and combined population. Then, 10 SNPs with lower P value in the combined population were selected from common SNPs among the first, second and combined populations. We developed algorithms using the 10 SNPs to predict effectiveness of infliximab and etanercept. In the validation population, availability of the algorithms was evaluated.

Results: Using combined infliximab-remission and LDA algorithms in the validation population, the accuracy was 90.4%. Using combined etanercept-remission and LDA algorithms in validation population, the accuracy was 76.8%.

Conclusions: The combined use of our two algorithms using SNPs may be very useful in the prediction of remission or LDA before treatment with infliximab or etanercept.

Purpose: Autofluorescence (AF) is the fluorescence of naturally occurring substances, such as nicotinamide adenine dinucleotide (NADH) and collagen. Two-photon microscopy (2PM) allows for the evaluation of living organs or tissues by excitation of tissue AF.

Methods: In the present study, we compared AF intensities of pancreatic tissues in different culture mediums [Roswell Park Memorial Institute (RPMI) 1640, 4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid (HEPES), and phosphate buffered saline (PBS) ] to determine the optimal conditions for observing unfixed and unstained tissues with 2PM.

Results: Tissues incubated in RPMI 1640 showed the highest overall fluorescence intensity, followed by those in HEPES and PBS. Comparing the fluorescence intensities of the respective wavelengths, two broad peaks (approximately 760 nm and 860 nm) were recognized. At approximately 760 nm, acinar cells and ductal cells were observed below the surface. This wavelength primarily detects NADH. At approximately 860 nm, dense fibrous tissue was observed on the pancreatic surface, suggesting the presence of a connective tissue surrounding the pancreas.

Conclusion: 2PM imaging using AF of the murine pancreas is a promising technique to provide new insights on structure and morphology.

Purpose: Rhythm exercise, such as walking, is effective in reducing stress associated with mental disorders, including anxiety and depression. Belt electrode skeletal muscle electrical stimulation (B-SES) can promote muscle contraction at a constant rhythm throughout the lower limbs without voluntary effort. B-SES may serve as a therapeutic strategy similar to rhythm exercise. The purpose of this study was to measure the effect of B-SES on stress on the basis of salivary amylase levels, a highly reliable stress index.

Methods: This study was a randomized controlled trial in which 30 healthy men were randomly assigned to three stimulation condition groups (DISUSE B-SES group, METABO B-SES group, and Control group). The salivary amylase levels were measured before, immediately after, and 30 minutes after the intervention. Changes in the salivary amylase levels were analyzed using repeated-measures analysis of variance.

Results: The METABO B-SES group had significantly different outcomes immediately after the intervention and 30 minutes after the end of the intervention.

Conclusion: METABO B-SES may contribute to stress reduction.

Background: Viral vector-mediated gene therapy has been eagerly studied as a new strategy for cancer treatment in recent years. We hypothesized that lentiviral vector-mediated gene therapy could be a promising strategy for the treatment of cancers that are highly refractory to chemotherapy or radiotherapy, such as malignant mesothelioma. In this study, we examined the effects of lentiviral vector-mediated transfection of tumor suppressor genes on the growth of malignant mesothelioma and other cancer cell lines.

Methods: We transfected the tumor suppressor genes p53, p16, and PTENinto various human cancer cell lines with lentiviral vectors.

Results and conclusion: After transfection of the tumor suppressor genes, we observed marked growth inhibition of the cancer cells. Thus, lentivirus-mediated transfection of tumor suppressor genes exerts promising anti-tumor effects on cancer cell lines.

Objectives: Neural tube defects (NTDs) are major congenital anomalies. To establish new and effective strategies to prevent these defects, public-private partnerships (PPPs) were formed between a healthcare company in Japan and numerous local governments to create educational activities regarding the importance of folic acid supplementation.

Methods: Projects for promoting maternal and child health using healthcare products and services created by a healthcare company were proposed to local governments as PPPs.

Results: Nine municipalities distributed folic acid supplements together with a Mother and Child Handbook while seven municipalities conducted surveys to identify people's knowledge and behavior toward folic acid use.

Conclusions: Recent studies have shown that periconceptional folic acid use improves many pregnancy outcomes; therefore, establishing educational activities regarding the importance of folic acid supplementation through PPPs is a rational and effective measure to promote maternal and child health.

Objectives: Measures by Public-Private Partnerships (PPPs) have been considered as possible modalities for local governments to conduct health promotion policies. The low energy meal replacement regimens are used for weight loss in obese patients. The aim of this project is to evaluate the feasibility and efficacy of a weight loss intervention program by a local government through PPPs in Japan.

Methods: Overweight or obese subjects in Sakai-town, Ibaraki pref., participated in the weight loss program, using telephone and/or personalized website to receive individual dietary counseling from medical professionals. Meal replacements were consumed once daily instead of regular meals. A standardized product contains 20.1 g of protein, one-third of the RDA of vitamins/minerals, and other food ingredients in 1 bag gross weight 50 g or 167 kcal. Anthropometric parameters and body composition were assessed before and after the 12-week weight-loss program.

Results: Ninety-two participants (male 47, female 45) completed the program between 1st, August 2018 and 13th, November 2018. Compared with the baseline, the significant decreases in BMI, body weight and waist circumference were observed.

Conclusion: The result demonstrated that a technology-based intervention, combined with meal replacements was feasible and effective for reducing body weight in obese subjects. The issues in implementing health promotion projects as PPPs will be examined, and further efforts to achieve the extension of the healthy lifespan will be taken to realize a healthy longevity society. The scheme of PPPs will play pivotal roles in the implementation of integrative health measure through public health services.

Purpose: Pin1 is a peptidyl prolyl cis/trans-isomerase (PPIase) that regulates phosphorylated protein function by cis-trans-isomerization. Pin1 catalytic activity has been associated with the pathogenesis of cancer, asthma, neurodegenerative diseases, nonalcoholic steatohepatitis, and viral infections. Recently, imidazole dipeptide molecules from natural products have attracted interest as functional food components. Here, we evaluated the effects of natural imidazole dipeptides on the PPIase activity of Pin1 to identify novel Pin1 inhibitors.

Methods: PPIase catalytic activity of Pin1 was measured using chymotrypsin-coupled isomer specific protein degradation. The test substance was preincubated with recombinant Pin1 to form their complex.

Results: Our results showed that among the imidazole dipeptide molecules, L-balenine, and not L-anserine and L-carnosine, reduced the PPIase activity of Pin1. Both β-alanine and 1-methyl-L-histidine obtained by hydrolysis of L-balenine did not affect Pin1 PPIase activity. These results suggested that the structure of L-balenine allows it to occupy the active site of the Pin1 enzyme.

Conclusion: Among imidazole dipeptides from natural products, L-balenine inhibited the PPIase catalytic activity of Pin1.

Cancer has been the leading cause of death in Japan since 1981, and deaths due to cancer continue to rise. The early detection of cancer and determining therapeutic effect using noninvasive techniques are critical in the treatment of cancer. We conducted a study assaying both circulating tumor cells (CTCs) and cell-free DNA (cfDNA) using a small quantity of blood (5 mL) in order to explore the utility of using liquid biopsies to detect and monitor various types of cancers. Our results confirm that not only are CTCs detected in multiple types of cancers, but also that there is a clinical correlation between the number of CTCs and cancer progression and the presence or absence of tumor metastasis. Furthermore, a significant increase in cfDNA concentration levels between healthy volunteers and cancer patients was confirmed. The measurements of CTCs and cfDNA levels have clinical significance, and can be expected to play a larger role in the diagnosis and treatment of cancer in the near future.