Systematic reduction of gray matter volume in anorexia nervosa, but relative enlargement with clinical symptoms in the prefrontal and posterior insular cortices: a multicenter neuroimaging study.

Although brain morphological abnormalities have been reported in anorexia nervosa (AN), the reliability and reproducibility of previous studies were limited due to insufficient sample sizes, which prevented exploratory analysis of the whole brain as opposed to regions of interest (ROIs). Objective was to identify brain morphological abnormalities in AN and the association with severity of AN by brain structural magnetic resonance imaging (MRI) in a multicenter study, and to conduct exploratory analysis of the whole brain. Here, we conducted a cross-sectional multicenter study using T1-weighted imaging (T1WI) data collected between May 2014 and February 2019 in Japan. We analyzed MRI data from 103 female AN patients (58 anorexia nervosa restricting type [ANR] and 45 anorexia nervosa binge-purging type [ANBP]) and 102 age-matched female healthy controls (HC). MRI data from five centers were preprocessed using the latest harmonization method to correct for intercenter differences. Gray matter volume (GMV) was calculated from T1WI data of all participants. Of the 205 participants, we obtained severity of eating disorder symptom scores from 179 participants, including 87 in the AN group (51 ANR, 36 ANBP) and 92 HC using the Eating Disorder Examination Questionnaire (EDE-Q) 6.0. GMV reduction were observed in the AN brain, including the bilateral cerebellum, middle and posterior cingulate gyrus, supplementary motor cortex, precentral gyrus medial segment, and thalamus. In addition, the orbitofrontal cortex (OFC), ventromedial prefrontal cortex (vmPFC), rostral anterior cingulate cortex (ACC), and posterior insula volumes showed positive correlations with severity of symptoms. This multicenter study was conducted with a large sample size to identify brain morphological abnormalities in AN. The findings provide a better understanding of the pathogenesis of AN and have potential for the development of brain imaging biomarkers of AN. Trial Registration: UMIN000017456. https://center6.umin.ac.jp/cgi-open-bin/icdr/ctr_view.cgi?recptno=R000019303 .

Comprehensive elucidation of resting-state functional connectivity in anorexia nervosa by a multicenter cross-sectional study.

BACKGROUND: Previous research on the changes in resting-state functional connectivity (rsFC) in anorexia nervosa (AN) has been limited by an insufficient sample size, which reduced the reliability of the results and made it difficult to set the whole brain as regions of interest (ROIs). METHODS: We analyzed functional magnetic resonance imaging data from 114 female AN patients and 135 healthy controls (HC) and obtained self-reported psychological scales, including eating disorder examination questionnaire 6.0. One hundred sixty-four cortical, subcortical, cerebellar, and network parcellation regions were considered as ROIs. We calculated the ROI-to-ROI rsFCs and performed group comparisons. RESULTS: Compared to HC, AN patients showed 12 stronger rsFCs mainly in regions containing dorsolateral prefrontal cortex (DLPFC), and 33 weaker rsFCs primarily in regions containing cerebellum, within temporal lobe, between posterior fusiform cortex and lateral part of visual network, and between anterior cingulate cortex (ACC) and thalamus (p < 0.01, false discovery rate [FDR] correction). Comparisons between AN subtypes showed that there were stronger rsFCs between right lingual gyrus and right supracalcarine cortex and between left temporal occipital fusiform cortex and medial part of visual network in the restricting type compared to the binge/purging type (p < 0.01, FDR correction). CONCLUSION: Stronger rsFCs in regions containing mainly DLPFC, and weaker rsFCs in regions containing primarily cerebellum, within temporal lobe, between posterior fusiform cortex and lateral part of visual network, and between ACC and thalamus, may represent categorical diagnostic markers discriminating AN patients from HC.

Association between somatosensory sensitivity and regional gray matter volume in healthy young volunteers: a voxel-based morphometry study.

Two-point discrimination (2PD) test reflects somatosensory spatial discrimination ability, but evidence on the relationship between 2PD and cortical gray matter (GM) volume is limited. This study aimed to analyze the relationship between cortical GM volume and 2PD threshold in young healthy individuals and to clarify the characteristics of brain structure reflecting the individual differences in somatosensory function. 2PD was measured in 42 healthy (20 females) volunteers aged 20-32 years using a custom-made test system that can be controlled by a personal computer. The 2PD of the right index finger measured with this device has been confirmed to show good reproducibility. T1-weighted images were acquired using a 3-T magnetic resonance imaging scanner for voxel-based morphometry analysis. The mean 2PD threshold was 2.58 ± 0.54 mm. Whole-brain multiple regression analysis of the relationship between 2PD and GM volume showed that a lower 2PD threshold (i.e. better somatosensory function) significantly correlated with decreased GM volume from the middle temporal gyrus to the inferior parietal lobule (IPL) in the contralateral hemisphere. In conclusion, a lower GM volume in the middle temporal gyrus and IPL correlates with better somatosensory function. Thus, cortical GM volume may be a biomarker of somatosensory function.

Gray Matter Volume Variability in Young Healthy Adults: Influence of Gender Difference and Brain-Derived Neurotrophic Factor Genotype.

Although brain gray matter (GM) plastically changes during short-term training, it is still unclear whether brain structures are stable for short periods (several months). Therefore, this study aimed to re-test the short-term variability of GM volumes and to clarify the effect of factors (gender and BDNF-genotype) expected to contribute to such variability. The subjects comprised 41 young healthy adults. T1-weighted images were acquired twice with an interval of approximately 4 months using a 3 T-MRI scanner. Voxel-based morphometry (VBM) was used to calculate GM volumes in 47 regions. The intraclass correlation coefficient (ICC) and Test-retest variability (%TRV) were used as indices of variability. As a result, the ICCs in 43 regions were excellent (ICC > 0.90) and those in 3 regions were good (ICC > 0.80), whereas the ICC in the thalamus was moderate (ICC = 0.694). Women had a higher %TRV than men in 5 regions, and %TRV of the Val66Val group was higher than that of the Met carrier group in 2 regions. Moreover, the Female-Val66Val group had a higher %TRV than the Male-Met carrier group in 3 regions. These results indicate that although the short-term variability of GM volumes is small, it is affected by within-subject factors.

Parafunctional masseter muscle activity during waking is related to periodontitis progression: A pilot prospective cohort study.

AIM: The purpose of this pilot prospective cohort study was to investigate the effects of parafunctional masseter muscle activity on periodontitis progression among patients receiving supporting periodontal therapy (SPT). MATERIALS AND METHODS: We collected data of patients treated at Okayama University Hospital from August 2014 to September 2018. The progression group was defined as patients with ≥2 teeth demonstrating a longitudinal loss of proximal attachment of ≥3 mm during the 3-year study period and/or at least one tooth extraction due to periodontitis progression. Surface electromyography of masseter muscles at baseline was continuously recorded while patients were awake and asleep. RESULTS: We analysed 48 patients (36 females) aged 66.8 ± 9.1 years (mean ± SD). The rate of parafunctional masseter muscle activity during waking hours and sleeping hours at baseline was 60.4% and 52.1%, respectively. Cox's proportional hazards regression model showed that the incidence of periodontitis progression was significantly associated with number of teeth present (p = 0.001) and parafunctional masseter muscle activity during waking hours (p = 0.041). CONCLUSION: Our results suggest that parafunctional masseter muscle activity during waking hours is a risk factor for periodontitis progression among patients receiving SPT.

Coordination of surface electromyography activity in the posterior tongue region during mastication of differently textured foods.

BACKGROUND: Masticatory movement occurs complicatedly and bilaterally. Although the tongue plays an important role in mastication, bilateral tongue function during mastication has not been clarified yet. OBJECTIVE: To investigate the effect of food properties on posterior tongue activity and coordination of muscles bilaterally by electromyography (EMG). METHODS: Twenty healthy adults (10 males and 10 females; mean age 28 years; range: 22-33 years) participated in this study. Three test foods, gummy jelly (hard food), sponge cake (soft food requiring crushing), and mashed potatoes (soft food not requiring crushing), were used. Bilateral masseter N-EMG (surface electromyography for measuring the muscle activity of posterior tongue) and submental EMG were carried out while the participants chewed three test foods. The participants were instructed to masticate three test foods only on the right side and only on the left side unilaterally. RESULTS: In the case of gummy jelly, N-EMG activity on the mastication side was significantly larger than that on the non-mastication side (P < .01). Regarding temporal relationship between the masseter and N-EMG activity, in the case of gummy jelly, the percentage of cases where the N-EMG peak was observed during masseter muscle EMG bursts was significantly higher than those for sponge cake and mashed potatoes (P < .01). CONCLUSION: N-EMG activity on the mastication side was significantly larger than that on the non-mastication side in the mastication of hard foods. Tongue showed activity pattern changes and coordinated with the masseter muscle depending on food texture.

Detection of muscle fatigue caused by repeated posterior tongue lift movement from neck surface EMG: a pilot study.

BACKGROUND: Fatigue is an important factor for muscle strengthening in rehabilitation medicine. Frequency analysis has been regarded as the gold standard for muscle fatigue assessment in surface electromyography (EMG). However, there are no experiments quantifying fatigue with grouped discharge (GD), which is one of the historical phenomena observed in patients having fatigue, by using high sampling rate recording of EMG. OBJECTIVE: To investigate the effect of fatigue, which is induced by repeated posterior tongue lift movement (TLM), on the occurrence of GD peaks, thus, to provide possible basis as a parameter for future fatigue evaluation. METHODS: Nineteen healthy adults (9 men and 10 women) participated in this study. The muscle fatigue protocol consisted of repetitive posterior TLM and maximum voluntary contractions (MVC). Bilateral N-EMG (Neck surface EMG for measuring the muscle activity of the posterior tongue) was recorded. RESULTS: Subjective tongue fatigue at the end of the protocol was significantly higher than at beginning throughout the muscle fatigue protocol (p < .01). The frequency of occurrence of GD peaks was 0.9 ± 1.2 (per second) at the baseline in 33 of 38 subject sides, and significantly increased depending on fatigue progression (p < .01). CONCLUSION: It was shown that fatigue due to repetitive posterior tongue lifting resulted in significant increase in the frequency of occurrence of GD peaks. Especially, the nature of GD, showing very few occurrences at non-fatigue condition, might work as an advantageous property for its use in future absolute evaluation of fatigue phenomena.

Effects of stage II transport and food bolus sampling methods on the properties of food particles.

During mastication, some portion of the food bolus is gradually transported to the oropharynx before deglutition, which is known as stage II transport (STII). Although the importance of STII in mastication and deglutition has been widely acknowledged, food particle properties that are transported into the oropharynx by STII have not yet been fully specified. To reveal the food particle properties that are transported into the oropharynx by STII and to assess the usefulness of new food bolus sampling methods for the evaluation of masticatory efficiency. Twenty-two healthy volunteers participated in this study. Four different bolus sampling conditions were adopted: (a) the whole food particles were expectorated when the subject was aware of the first deglutition, (b) the last bolus to be swallowed at the end of unrestrained food intake was expectorated (aftermost bolus sample), (3) the whole food particles were expectorated when the subject felt ready to swallow after swallow-inhibited mastication (swallow-inhibited sample), (4) the particles were regurgitated from the oropharynx after the first STII (stage II-transported sample). Food particles were analysed using the homogeneity index and particle size index. There was no significant difference between food particles in the aftermost bolus sample and swallow-inhibited sample. The particles in the stage II-transported sample showed significantly more homogeneous and smaller sizes than other sampling conditions (P < .05). The food particles transferred to the oropharynx in the stage II-transported sample were smaller than those broken down by natural mastication.

A within-subject clinical trial on the conversion of mandibular two-implant to three-implant overdenture: Patient-centered outcomes and willingness to pay.

OBJECTIVES: To examine the impact of adding a third midline implant with stud attachment to a mandibular two-implant overdenture on patient-oriented outcomes. METHODS: In this pre-post design clinical trial, following the standard procedures, mandibular two-implant overdentures of 17 edentulous individuals (61.9 ± 6.6 years) were converted to three-implant overdentures by adding a stud attachment to an unloaded midline implant. Patient-oriented outcomes included patient expectations and satisfaction with implant overdenture as well as willingness to pay the cost of conversion. Data were collected at baseline and at the 6-week follow-up using visual analog and binary scales as well as open-ended questions. Statistical analysis included descriptive statistics, Spearman's correlation, Fisher exact test, Mann-Whitney U test, and the exact sign test. RESULTS: After connecting the third midline implant to the mandibular two-implant overdenture, there was a statistically significant decrease in the anteroposterior movement (p = 0.005) as evaluated by clinicians. Moreover, study participants reported an increase in perceived stability of the overdenture (95% CI; 0.68-1.00, p = 0.002) and in their ability to speak (95% CI; 0.63-1.00, p = 0.008). The addition of a third implant met the expectations of 94% of patients in regard to lower denture stability, 100% for retention, and 82.4% for comfort. The mandibular three-implant overdenture increased patient general satisfaction over a short period of time, but this improvement was not statistically significant. About 80% of patients would recommend this type of prosthesis to their peers but only 47% of them would agree to pay a large increase in the cost of treatment compared to a two-implant overdenture. CONCLUSIONS: The addition of a midline third implant to an existing mandibular two-implant overdenture resulted in several improved patient-reported outcomes.

Characteristics of Grouped Discharge Waveforms Observed in Long-term Masseter Muscle Electromyographic Recording: A Preliminary Study.

We investigated the characteristics of grouped discharge (GD) waveforms obtained from long-term masseter electromyogram (EMG) recording in 6 female temporomandibular disorder (TMD) patients with myofascial pain and 6 healthy females. The EMG measurement was performed from the morning of the experiment day until the subject woke up the next day. We observed a significantly larger number of GD waveforms in the TMD group compared to the control group (p=0.002). Our results indicate that the existence of GD waveforms in masseter EMGs might be a predictor of future TMD with myofascial pain.

Effect of food properties and chewing condition on the electromyographic activity of the posterior tongue.

BACKGROUND: Retraction of the posterior tongue facilitates bolus formation, food transfer and Stage II transport in chewing and swallowing. Surface electromyography of the neck region (N-EMG) enabled the evaluation of muscle function at the posterior tongue. Although previous studies showed that food hardness increased masticatory muscles EMG activity and volitional chewing altered swallowing, the effect of food properties and chewing condition on N-EMG activity has not yet been elucidated. OBJECTIVE: To investigate the effect of food properties and chewing conditions on N-EMG. METHODS: Twelve healthy adults (six men and six women, mean age 28.1 ± 2.7) participated in this study. Three test foods were used: a cookie (solid food), crushed cookies adjusted using agar and water (semisolid food) and crushed cookies adjusted using thickening agent and water (soft food). Masseter and submental muscle EMG and N-EMG activity were recorded while chewing the three test foods. Participants were instructed to masticate the three test foods in three chewing conditions: (a) chewing freely, (b) chewing test foods with the same number of chewing cycles as when chewing solid food and (c) chewing with a maximum effort. RESULTS: Total N-EMG activity when chewing solid food was higher than that in the other two test foods regardless of the chewing condition. In soft food and semisolid food, the total N-EMG activity was increased by increasing the number of chewing cycles. CONCLUSION: The total N-EMG activity was increased while chewing hard food, which indicates the positive effect of chewing hard food on rehabilitation at the posterior tongue.

A new posture measurement method for measuring intrinsic standing posture.

AIMS: Although body posture in relation to the dental condition has been of great interest in the dental profession, rumination bias has been a substantial obstacle to achieving a reliable objective evaluation of the intrinsic body posture. The aim of this study was to establish a posture control protocol that would minimize the effect of bias. METHODOLOGY: Fifteen healthy male volunteers (23-33 years of age) participated in this study. The posture movement was recorded for 10 seconds by a three-dimensional motion capture system. The experiment was performed on four different days. RESULTS: The posture was most stable at 4-5 seconds after the start of the front bulb gaze (the mean coefficient of variation ranged from 0.1 to 44.1). The intraclass correlation coefficients for four days were 0.871-0.975 (P < or = 0.001). CONCLUSIONS: It was concluded that the use of this measurement method helped in producing a reliable intrinsic standing posture where unbiased evaluation of the effect of any intervention on the body posture is researched.

[A Study on the Uncertainty of the Average Glandular Dose in Quality Control of Mammographic X-ray Units].

【Purpose】 Accurate control of X-ray units and dosimeters and analysis of the uncertainties associated with the accurate measurement of radiation doses are essential for the effective establishment and application of diagnostic reference levels. In this study, the uncertainty of the average glandular dose (AGD) in the quality control of mammography equipment was evaluated in detail, and recommendations were provided to improve the accuracy and safety of radiological practice. 【Methods】 In the uncertainty analysis of the AGD, the relative standard uncertainties in the measurements of the half-value layer, the incident air kerma, and the conversion factor were considered and finally expressed as expanded uncertainties, the intervals of which were clearly defined. 【Results】 From the AGD measurements using two types of dosimeters, it was found that the primary sources of uncertainty are the uncertainty of the calibration factors of the dosimeters and the uncertainty of the conversion factors.【Conclusion】 To reduce uncertainty, the use of regularly calibrated dosimeters is effective and reliable. Two types of dosimeters are commonly used; the results of this study may serve as a reference value for the uncertainty of AGD in quality control in medical facilities.

Identification of areas of the brain activated by active stimulation in hairless skin.

In the past few decades, neuroscientists have studied the physiological basis of pleasant touch. Unmyelinated low-threshold mechanoreceptors are central to the study of the physiological basis of pleasant touch. Research on pleasant stimuli has mostly focused on passive stimuli, and the brain activation sites for active pleasant stimuli are not clear. Therefore, the purpose of this study was to identify brain activation sites during active pleasant stimulation of hairless skin using functional magnetic resonance imaging. Forty-two healthy subjects aged 19 years or older were asked to actively grasp in five stimulus tasks. The comfort and sensations that occurred during the tasks were investigated using a questionnaire. Significant activation was found in the middle frontal gyrus when the hair ball and slime ball were grasped, while there was significant activation in the amygdala when grasping a squeeze ball compared to the tennis ball. In a questionnaire survey of the subjects, there was a significant difference in the comfort score between the tennis ball and the squeeze ball, but no significant correlation was found between the comfort scores and the brain sites of activation. Therefore, although active stimulation with the squeeze ball significantly activated the amygdala, it was not clear that the amygdala was significantly activated by active pleasant stimulation. In the future, it will be necessary to investigate the texture of the squeeze ball in more detail, and to increase the number of subjects for further study.

Construction and evaluation of a neurofeedback system using finger tapping and near-infrared spectroscopy.

Introduction: Neurofeedback using near-infrared spectroscopy (NIRS) has been used in patients with stroke and other patients, but few studies have included older people or patients with cognitive impairment. Methods: We constructed a NIRS-based neurofeedback system and used finger tapping to investigate whether neurofeedback can be implemented in older adults while finger tapping and whether brain activity improves in older adults and healthy participants. Our simple neurofeedback system was constructed using a portable wearable optical topography (WOT-HS) device. Brain activity was evaluated in 10 older and 31 healthy young individuals by measuring oxygenated hemoglobin concentration during finger tapping and neurofeedback implementation. Results: During neurofeedback, the concentration of oxygenated hemoglobin increased in the prefrontal regions in both the young and older participants. Discussion: The results of this study demonstrate the usefulness of neurofeedback using simple NIRS devices for older adults and its potential to mitigate cognitive decline.

Identifying factors that indicate the possibility of non-visible cases on mammograms using mammary gland content ratio estimated by artificial intelligence.

Background: Mammography is the modality of choice for breast cancer screening. However, some cases of breast cancer have been diagnosed through ultrasonography alone with no or benign findings on mammography (hereby referred to as non-visibles). Therefore, this study aimed to identify factors that indicate the possibility of non-visibles based on the mammary gland content ratio estimated using artificial intelligence (AI) by patient age and compressed breast thickness (CBT). Methods: We used AI previously developed by us to estimate the mammary gland content ratio and quantitatively analyze 26,232 controls and 150 non-visibles. First, we evaluated divergence trends between controls and non-visibles based on the average estimated mammary gland content ratio to ensure the importance of analysis by age and CBT. Next, we evaluated the possibility that mammary gland content ratio ≥50% groups affect the divergence between controls and non-visibles to specifically identify factors that indicate the possibility of non-visibles. The images were classified into two groups for the estimated mammary gland content ratios with a threshold of 50%, and logistic regression analysis was performed between controls and non-visibles. Results: The average estimated mammary gland content ratio was significantly higher in non-visibles than in controls when the overall sample, the patient age was ≥40 years and the CBT was ≥40 mm (p < 0.05). The differences in the average estimated mammary gland content ratios in the controls and non-visibles for the overall sample was 7.54%, the differences in patients aged 40-49, 50-59, and ≥60 years were 6.20%, 7.48%, and 4.78%, respectively, and the differences in those with a CBT of 40-49, 50-59, and ≥60 mm were 6.67%, 9.71%, and 16.13%, respectively. In evaluating mammary gland content ratio ≥50% groups, we also found positive correlations for non-visibles when controls were used as the baseline for the overall sample, in patients aged 40-59 years, and in those with a CBT ≥40 mm (p < 0.05). The corresponding odds ratios were ≥2.20, with a maximum value of 4.36. Conclusion: The study findings highlight an estimated mammary gland content ratio of ≥50% in patients aged 40-59 years or in those with ≥40 mm CBT could be indicative factors for non-visibles.

Development of a Subtraction Processing Technology for Assistance in the Comparative Interpretation of Mammograms.

A comparative interpretation of mammograms has become increasingly important, and it is crucial to develop subtraction processing and registration methods for mammograms. However, nonrigid image registration has seldom been applied to subjects constructed with soft tissue only, such as mammograms. We examined whether subtraction processing for the comparative interpretation of mammograms can be performed using nonrigid image registration. As a preliminary study, we evaluated the results of subtraction processing by applying nonrigid image registration to normal mammograms, assuming a comparative interpretation between the left and right breasts. Mediolateral-oblique-view mammograms were taken from noncancer patients and divided into 1000 cases for training, 100 cases for validation, and 500 cases for testing. Nonrigid image registration was applied to align the horizontally flipped left-breast mammogram with the right one. We compared the sum of absolute differences (SAD) of the difference of bilateral images (Difference Image) with and without the application of nonrigid image registration. Statistically, the average SAD was significantly lower with the application of nonrigid image registration than without it (without: 0.0692; with: 0.0549 (p < 0.001)). In four subgroups using the breast area, breast density, compressed breast thickness, and Difference Image without nonrigid image registration, the average SAD of the Difference Image was also significantly lower with nonrigid image registration than without it (p < 0.001). Nonrigid image registration was found to be sufficiently useful in aligning bilateral mammograms, and it is expected to be an important tool in the development of a support system for the comparative interpretation of mammograms.

PD-L1 expression in keratinocyte and infiltration of CD4 + T lymphocyte can predict a severe type of erythema multiforme major induced by the anti-PD-1 antibody, pembrolizumab.

Skin toxicity is the most common adverse event of treatment with immune check point inhibitors. Among them, erythema multiforme is a rare occurrence with a frequency of 4%, with most of the cases developing grade 1/2 disease. We experienced high grade erythema multiforme major developing with pembrolizumab treatment for anal canal cancer with extensive skin metastases. Steroid ointment was ineffective, and the skin lesions with blisters expanded to > 45% of the body surface area. The patient was at risk for symptom aggravation, and a pulse therapy with methylprednisolone and increasing the dose of oral prednisolone (1 mg/kg) were started. The skin lesions improved in 1.8 months. Unless urgent and appropriate treatments such as high dose steroid administration were conducted, the skin toxicities could not be controlled. The presence of CD4+ T cells and PD-L1+ keratinocytes in the skin biopsy might be a predictive marker of erythema multiforme major resistant to standard steroid treatment. Supplementary Information: The online version contains supplementary material available at 10.1007/s13691-024-00676-4.

The gut lactic acid bacteria metabolite, 10-oxo-cis-6,trans-11-octadecadienoic acid, suppresses inflammatory bowel disease in mice by modulating the NRF2 pathway and GPCR-signaling.

Various gut bacteria, including Lactobacillus plantarum, possess several enzymes that produce hydroxy fatty acids (FAs), oxo FAs, conjugated FAs, and partially saturated FAs from polyunsaturated FAs as secondary metabolites. Among these derivatives, we identified 10-oxo-cis-6,trans-11-octadecadienoic acid (γKetoC), a γ-linolenic acid (GLA)-derived enon FA, as the most effective immunomodulator, which inhibited the antigen-induced immunoactivation and LPS-induced production of inflammatory cytokines. The treatment with γKetoC significantly suppressed proliferation of CD4+ T cells, LPS-induced activation of bone marrow-derived dendritic cells (BMDCs), and LPS-induced IL-6 release from peritoneal cells, splenocytes, and CD11c+ cells isolated from the spleen. γKetoC also inhibited the release of inflammatory cytokines from BMDCs stimulated with poly-I:C, R-848, or CpG. Further in vitro experiments using an agonist of GPR40/120 suggested the involvement of these GPCRs in the effects of γKetoC on DCs. We also found that γKetoC stimulated the NRF2 pathway in DCs, and the suppressive effects of γKetoC and agonist of GPR40/120 on the release of IL-6 and IL-12 were reduced in Nrf2-/- BMDCs. We evaluated the role of NRF2 in the anti-inflammatory effects of γKetoC in a dextran sodium sulfate-induced colitis model. The oral administration of γKetoC significantly reduced body weight loss, improved stool scores, and attenuated atrophy of the colon, in wild-type C57BL/6 and Nrf2+/- mice with colitis. In contrast, the pathology of colitis was deteriorated in Nrf2-/- mice even with the administration of γKetoC. Collectively, the present results demonstrated the involvement of the NRF2 pathway and GPCRs in γKetoC-mediated anti-inflammatory responses.

ITGA3 and ITGB4 expression biomarkers estimate the risks of locoregional and hematogenous dissemination of oral squamous cell carcinoma.

BACKGROUND: Molecular biomarkers are essential for monitoring treatment effects, predicting prognosis, and improving survival rate in oral squamous cell carcinoma. This study sought to verify the effectiveness of two integrin gene expression ratios as biomarkers. METHODS: Gene expression analyses of integrin α3 (ITGA3), integrin ß4 (ITGB4), CD9 antigen (CD9), and plakoglobin (JUP) by quantitative real-time PCR were conducted on total RNA from 270 OSCC cases. The logrank test, Cox proportional hazards model, and Kaplan-Meier estimates were performed on the gene expression ratios of ITGA3/CD9 and ITGB4/JUP and on the clinicopathological parameters for major clinical events. RESULTS: A high rate (around 80%) of lymph node metastasis was found in cases with a high ITGA3/CD9 ratio (high-ITGA3/CD9) and invasive histopathology (YK4). Primary site recurrence (PSR) was associated with high-ITGA3/CD9, T3-4 (TNM class), and positive margin, indicating that PSR is synergistically influenced by treatment failure and biological malignancy. A high ITGB4/JUP ratio (high-ITGB4/JUP) was revealed to be a primary contributor to distant metastasis without the involvement of clinicopathological factors, suggesting intervention of a critical step dependent on the function of the integrin ß4 subunit. Kaplan-Meier curves revealed positive margin as a lethal treatment consequence in high-ITGA3/CD9 and YK4 double-positive cases. CONCLUSION: Two types of metastatic trait were found in OSCC: locoregional dissemination, which was reflected by high-ITGA3/CD9, and distant metastasis through hematogenous dissemination, uniquely distinguished by high-ITGB4/JUP. The clinical significance of the integrin biomarkers implies that biological mechanisms such as cancer cell motility and anchorage-independent survival are vital for OSCC recurrence and metastasis.