A pilot randomized clinical trial of biomedical link with mental health in art therapy intervention programs for alcohol use disorder: Changes in NK cells, addiction biomarkers, electroencephalography, and MMPI-2 profiles.

OBJECTIVE: Alcohol intake is a major risk factor for various diseases. Elucidating alcohol use disorder (AUD) is important in preventing diseases and promoting health. We aimed to investigate the effect of art therapy on emotional (Minnesota Multiphasic Personality Inventory-2 [MMPI-2]) and physical (natural killer [NK] cell count, expression of stress-associated proteins [SAP], and electroencephalography) changes in patients with AUD. METHODS: Participants were randomly divided into two groups (n = 35), with the experimental group undergoing art therapy involving weekly 60-min group therapy sessions for 10 weeks. Statistical analysis was performed using Ranked ANCOVA and Wilcoxon's signed rank test. Western blotting was performed to analyze serum SAP levels. RESULTS: We observed an association between psychological mechanisms and stress proteins. There was an increased number of NK cells in the experimental group after the program. Moreover, compared with the control group, the experimental group showed significant changes in SAP expression. Further, the experimental group showed a positive change in the MMPI-2 profile, as well as a decrease in depression, anxiety, impulsivity, and alcohol dependence. CONCLUSIONS: Continuous psychological support could be applied as a stress-control program for preventing stress recurrence and post-discharge relapse. Our findings strengthen the link between biomedical science and mental health in rehabilitation treatment for AUD.

Transcriptome Analysis Reveals Immunomodulatory Effect of Spore-Displayed p75 on Human Intestinal Epithelial Caco-2 Cells.

Lacticaseibacillus rhamnosus GG (LGG) can promote intestinal health by modulating the immune responses of the gastrointestinal tract. However, knowledge about the immunomodulatory action of LGG-derived soluble factors is limited. In our previous study, we have displayed LGG-derived p75 protein on the spore surface of Bacillus subtilis. The objective of this study was to determine the effect of spore-displayed p75 (CotG-p75) on immune system by investigating transcriptional response of Caco-2 cells stimulated by CotG-p75 through RNA-sequencing (RNA-seq). RNA-seq results showed that CotG-p75 mainly stimulated genes involved in biological processes, such as response to stimulus, immune regulation, and chemotaxis. KEGG pathway analysis suggested that many genes activated by CotG-p75 were involved in NF-ĸB signaling and chemokine signaling pathways. CotG-p75 increased cytokines and chemokines such as CXCL1, CXCL2, CXCL3, CXCL8, CXCL10, CCL20, CCL22, and IL1B essential for the immune system. In particular, CotG-p75 increased the expression levels of NF-ĸB-related genes such as NFKBIA, TNFAIP3, BIRC3, NFKB2, and RELB involved in immune and inflammatory responses. This study provides genes and pathways involved in immune responses influenced by CotG-p75. These comprehensive transcriptome profiling could be used to elucidate the immunomodulatory action of CotG-p75.

Transcriptional responses of human intestinal epithelial HT-29 cells to spore-displayed p40 derived from Lacticaseibacillus rhamnosus GG.

BACKGROUNDS: The aims of this study were to construct spore-displayed p40, a Lacticaseibacillus rhamnosus GG-derived soluble protein, using spore surface display technology and to evaluate transcriptional responses in human intestinal epithelial cells. RESULTS: p40 was displayed on the surface of Bacillus subtilis spores using spore coat protein CotG as an anchor protein. Effects of spore-displayed p40 (CotG-p40) on gene expression of intestinal epithelial cell line HT-29 were evaluated by transcriptome analysis using RNA-sequencing. As a result of differentially expressed gene (DEG) analysis, 81 genes were up-regulated and 82 genes were down-regulated in CotG-p40 stimulated cells than in unstimulated cells. Gene ontology enrichment analysis showed that CotG-p40 affected biological processes such as developmental process, metabolic process, cell surface receptor linked signaling pathway, and retinoic acid metabolic process. Gene-gene network analysis suggested that 10 DEGs (EREG, FOXF1, GLI2, PTGS2, SPP1, MMP19, TNFRSF1B, PTGER4, CLDN18, and ALDH1A3) activated by CotG-p40 were associated with probiotic action. CONCLUSIONS: This study demonstrates the regulatory effects of CotG-p40 on proliferation and homeostasis of HT-29 cells. This study provided comprehensive insights into the transcriptional response of human intestinal epithelial cells stimulated by CotG-p40.

Bioactive Compounds and Quality Evaluation of Red-Pigmented Rice Processed by Germination and Roasting.

Red-pigmented rice was germinated and processed to develop germinated red rice tea, and the changes in physicochemical, bioactive, and microbial properties due to germination and roasting were investigated. The moisture and crude ash contents of red rice decreased after germination and roasting. Crude protein and crude fat contents increased after germination but slightly decreased after roasting. Total phenolics, flavonoids, and antioxidant activities (DPPH and ABTS radical scavenging activities) increased following germination and heat treatment. However, the increased levels of γ-amino butyric acid after germination significantly decreased during the subsequent roasting step. In addition, total bacteria, yeast, and mold counts increased during the germination process but decreased after heat treatment as compared to those in the original grain; Escherichia coli was not detected. Therefore, germination and subsequent roasting could effectively enhance the contents of the most bioactive compounds and maintain microbial stability in red-pigmented rice.

Effects of Spore-Displayed p75 Protein from Lacticaseibacillus rhamnosus GG on the Transcriptional Response of HT-29 Cells.

A Lacticaseibacillus rhamnosus GG-derived protein, p75, is one of the key molecules exhibiting probiotic activity. However, the molecular mechanism and transcriptional response of p75 in human intestinal epithelial cells are not completely understood. To gain a deeper understanding of its potential probiotic action, this study investigated genome-wide responses of HT-29 cells to stimulation by spore-displayed p75 (CotG-p75) through a transcriptome analysis based on RNA sequencing. Analysis of RNA-seq data showed significant changes of gene expression in HT-29 cells stimulated by CotG-p75 compared to the control. A total of 189 up-regulated and 314 down-regulated genes was found as differentially expressed genes. Gene ontology enrichment analysis revealed that a large number of activated genes was involved in biological processes, such as epithelial cell differentiation, development, and regulation of cell proliferation. A gene-gene interaction network analysis showed that several DEGs, including AREG, EREG, HBEGF, EPGN, FASLG, GLI2, CDKN1A, FOSL1, MYC, SERPINE1, TNFSF10, BCL6, FLG, IVL, SPRR1A, SPRR1B, SPRR3, and MUC5AC, might play a critical role in these biological processes. RNA-seq results for selected genes were verified by reverse transcription-quantitative polymerase chain reaction. Overall, these results provide extensive knowledge about the transcriptional responses of HT-29 cells to stimulation by CotG-p75. This study showed that CotG-p75 can contribute to cell survival and epithelial development in human intestinal epithelial cells.

Effects of Nature-Based Group Art Therapy Programs on Stress, Self-Esteem and Changes in Electroencephalogram (EEG) in Non-Disabled Siblings of Children with Disabilities.

The purpose of the present study was to examine changes in brain waves, stress, and self-esteem after a continuous eight-week nature-based art therapy program in the forest in non-disabled siblings of children with disabilities. A total of 29 participants participated in this study (art therapy program group, n = 18; control group, n = 11). The art therapy program group received eight weekly sessions of art therapy lasting 60 min each. Pre- and post-test results showed positive changes in the brain function index and stress levels of the participants in the art therapy program group. On the self-esteem scale, overall and social self-esteem increased significantly. In conclusion, creative activities in the forest can increase resistance to diseases through mechanisms that relieve stress and increase self-esteem. If art therapy that emphasizes somatosensory experience, creative expression, and self-motivation is accompanied by forest activities, this combined intervention can elicit positive physical and psychological changes.

Physicochemical and bioactive properties of a high ß-glucan barley variety 'Betaone' affected by germination processing.

This study was conducted to investigate the effects of germination on the bioactive phytochemicals in the barley variety 'Betaone', which has high ß-glucan content. Betaone barley seeds were germinated for 24, 48, 72, and 96 h at 25 °C in a growth chamber. As the germination period progressed, crude protein, crude fat, and crude ash levels increased, while starch content decreased. ß-Glucan content significantly decreased during the germination period. However, after 96 h of germination, it was still 4.31%, which is similar to the amount found in ordinary barley varieties. The α- and ß-amylase activities in Betaone barley increased, thereby significantly decreasing pasting viscosities (p ≤ 0.05), as germination progressed. The water absorption index increased in the early germination stage, subsequently decreasing as germination progressed. While the water solubility index fell in the first stage of germination, it subsequently increased as germination progressed. Oil absorption decreased as germination progressed. Bioactive compounds, in particular the total phenolic (122.84-322.67 µg/g), total flavonoid (32.20-124.09 µg/g), and GABA content (176.94-212.64 µg/g), increased as germination progressed. The antioxidative properties mainly DPPH and ABTS radical scavenging activity also increased during the germination period. Therefore, these findings could be a useful method for improving new products with enhanced bioactive phytochemicals.

Comparison of Seven Commercial TaqMan Master Mixes and Two Real-Time PCR Platforms Regarding the Rapid Detection of Porcine DNA.

A pig-specific real-time PCR assay based on the mitochondrial ND5 gene was developed to detect porcine material in food and other products. To optimize the performance of assay, seven commercial TaqMan master mixes and two real-time PCR platforms (Applied Biosystems StepOnePlus and Bio-rad CFX Connect) were used to evaluate the limit of detection (LOD) as well as the PCR efficiency and specificity. The LODs and PCR efficiencies for the seven master mixes on two platforms were 0.5-5 pg/reaction and 84.96%-108.80%, respectively. Additionally, non-specific amplifications of DNA from other animal samples (human, dog, cow, and chicken) were observed for four master mixes. These results imply that the sensitivity and specificity of a real-time PCR assay may vary depending on master mix and platform used. The best combination of master mix and real-time PCR platform can accurately detect 0.5 pg porcine DNA, with a PCR efficiency of 100.49%.

Surface display of p75, a Lactobacillus rhamnosus GG derived protein, on Bacillus subtilis spores and its antibacterial activity against Listeria monocytogenes.

Lactobacillus rhamnosus p75 protein with peptidoglycan hydrolase (PGH) activity is one of the key molecules exhibiting anti-apoptotic and cell-protective activity for human intestinal epithelial cells. In this study, with the goal of developing new probiotics, the p75 protein was displayed on the surface of Bacillus subtilis spores using spore coat protein CotG as an anchoring motif. The PGH activity, stability, and the antibacterial activity of the spore-displayed p75 (CotG-p75) protein were also investigated. The PGH activity of the CotG-p75 against peptidoglycan extracted from B. subtilis was confirmed by the ninhydrin test. Under various harsh conditions, compared to the control groups, the PGH activities of CotG-p75 were very stable in the range of pH 3-7 and maintained at 70% at 50 °C. In addition, the antibacterial activity of CotG-p75 against Listeria monocytogenes was evaluated by a time-kill assay. After 6 h incubation in phosphate-buffered saline, CotG-p75 reduced the number of viable cells of L. monocytogenes by up to 2.0 log. Scanning electron microscopy analysis showed that the cell wall of L. monocytogenes was partially damaged by the treatment with CotG-p75. Our preliminary results show that CotG-p75 could be a good candidate for further research to develop new genetically engineered probiotics.