TREK-1 and Best1 channels mediate fast and slow glutamate release in astrocytes upon GPCR activation.

Astrocytes release glutamate upon activation of various GPCRs to exert important roles in synaptic functions. However, the molecular mechanism of release has been controversial. Here, we report two kinetically distinct modes of nonvesicular, channel-mediated glutamate release. The fast mode requires activation of G(αi), dissociation of G(ßγ), and subsequent opening of glutamate-permeable, two-pore domain potassium channel TREK-1 through direct interaction between G(ßγ) and N terminus of TREK-1. The slow mode is Ca(2+) dependent and requires G(αq) activation and opening of glutamate-permeable, Ca(2+)-activated anion channel Best1. Ultrastructural analyses demonstrate that TREK-1 is preferentially localized at cell body and processes, whereas Best1 is mostly found in microdomains of astrocytes near synapses. Diffusion modeling predicts that the fast mode can target neuronal mGluR with peak glutamate concentration of 100 µM, whereas slow mode targets neuronal NMDA receptors at around 1 µM. Our results reveal two distinct sources of astrocytic glutamate that can differentially influence neighboring neurons.

MST1 Negatively Regulates TNFα-Induced NF-κB Signaling through Modulating LUBAC Activity.

The nuclear factor (NF)-κB pathway plays a central role in inflammatory and immune responses, with aberrant activation of NF-κB signaling being implicated in various human disorders. Here, we show that mammalian ste20-like kinase 1 (MST1) is a previously unrecognized component of the tumor necrosis factor α (TNFα) receptor 1 signaling complex (TNF-RSC) and attenuates TNFα-induced NF-κB signaling. Genetic ablation of MST1 in mouse embryonic fibroblasts and bone marrow-derived macrophages potentiated the TNFα-induced increase in IκB kinase (IKK) activity, as well as the expression of NF-κB target genes. TNFα induced the recruitment of MST1 to TNF-RSC and its interaction with HOIP, the catalytic component of the E3 ligase linear ubiquitin assembly complex (LUBAC). Furthermore, MST1 activated in response to TNFα stimulation mediates the phosphorylation of HOIP and thereby inhibited LUBAC-dependent linear ubiquitination of NEMO/IKKγ. Together, our findings suggest that MST1 negatively regulates TNFα-induced NF-κB signaling by targeting LUBAC.

Prefoldin 6 mediates longevity response from heat shock factor 1 to FOXO in C. elegans.

Heat shock factor 1 (HSF-1) and forkhead box O (FOXO) are key transcription factors that protect cells from various stresses. In Caenorhabditis elegans, HSF-1 and FOXO together promote a long life span when insulin/IGF-1 signaling (IIS) is reduced. However, it remains poorly understood how HSF-1 and FOXO cooperate to confer IIS-mediated longevity. Here, we show that prefoldin 6 (PFD-6), a component of the molecular chaperone prefoldin-like complex, relays longevity response from HSF-1 to FOXO under reduced IIS. We found that PFD-6 was specifically required for reduced IIS-mediated longevity by acting in the intestine and hypodermis. We showed that HSF-1 increased the levels of PFD-6 proteins, which in turn directly bound FOXO and enhanced its transcriptional activity. Our work suggests that the prefoldin-like chaperone complex mediates longevity response from HSF-1 to FOXO to increase the life span in animals with reduced IIS.

The circadian E3 ligase FBXL21 regulates myoblast differentiation and sarcomere architecture via MYOZ1 ubiquitination and NFAT signaling.

Numerous molecular and physiological processes in the skeletal muscle undergo circadian time-dependent oscillations in accordance with daily activity/rest cycles. The circadian regulatory mechanisms underlying these cyclic processes, especially at the post-transcriptional level, are not well defined. Previously, we reported that the circadian E3 ligase FBXL21 mediates rhythmic degradation of the sarcomere protein TCAP in conjunction with GSK-3ß, and Psttm mice harboring an Fbxl21 hypomorph allele show reduced muscle fiber diameter and impaired muscle function. To further elucidate the regulatory function of FBXL21 in skeletal muscle, we investigated another sarcomere protein, Myozenin1 (MYOZ1), that we identified as an FBXL21-binding protein from yeast 2-hybrid screening. We show that FBXL21 binding to MYOZ1 led to ubiquitination-mediated proteasomal degradation. GSK-3ß co-expression and inhibition were found to accelerate and decelerate FBXL21-mediated MYOZ1 degradation, respectively. Previously, MYOZ1 has been shown to inhibit calcineurin/NFAT signaling important for muscle differentiation. In accordance, Fbxl21 KO and MyoZ1 KO in C2C12 cells impaired and enhanced myogenic differentiation respectively compared with control C2C12 cells, concomitant with distinct effects on NFAT nuclear localization and NFAT target gene expression. Importantly, in Psttm mice, both the levels and diurnal rhythm of NFAT2 nuclear localization were significantly diminished relative to wild-type mice, and circadian expression of NFAT target genes associated with muscle differentiation was also markedly dampened. Furthermore, Psttm mice exhibited significant disruption of sarcomere structure with a considerable excess of MYOZ1 accumulation in the Z-line. Taken together, our study illustrates a pivotal role of FBXL21 in sarcomere structure and muscle differentiation by regulating MYOZ1 degradation and NFAT2 signaling.

Regulation of headache response and transcriptomic network by the trigeminal ganglion clock.

OBJECTIVE: To characterize the circadian features of the trigeminal ganglion in a mouse model of headache. BACKGROUND: Several headache disorders, such as migraine and cluster headache, are known to exhibit distinct circadian rhythms of attacks. The circadian basis for these rhythmic pain responses, however, remains poorly understood. METHODS: We examined trigeminal ganglion ex vivo and single-cell cultures from Per2::LucSV reporter mice and performed immunohistochemistry. Circadian behavior and transcriptomics were investigated using a novel combination of trigeminovascular and circadian models: a nitroglycerin mouse headache model with mechanical thresholds measured every 6 h, and trigeminal ganglion RNA sequencing measured every 4 h for 24 h. Finally, we performed pharmacogenomic analysis of gene targets for migraine, cluster headache, and trigeminal neuralgia treatments as well as trigeminal ganglion neuropeptides; this information was cross-referenced with our cycling genes from RNA sequencing data to identify potential targets for chronotherapy. RESULTS: The trigeminal ganglion demonstrates strong circadian rhythms in both ex vivo and single-cell cultures, with core circadian proteins found in both neuronal and non-neuronal cells. Using our novel behavioral model, we showed that nitroglycerin-treated mice display circadian rhythms of pain sensitivity which were abolished in arrhythmic Per1/2 double knockout mice. Furthermore, RNA-sequencing analysis of the trigeminal ganglion revealed 466 genes that displayed circadian oscillations in the control group, including core clock genes and clock-regulated pain neurotransmitters. In the nitroglycerin group, we observed a profound circadian reprogramming of gene expression, as 331 of circadian genes in the control group lost rhythm and another 584 genes gained rhythm. Finally, pharmacogenetics analysis identified 10 genes in our trigeminal ganglion circadian transcriptome that encode target proteins of current medications used to treat migraine, cluster headache, or trigeminal neuralgia. CONCLUSION: Our study unveiled robust circadian rhythms in the trigeminal ganglion at the behavioral, transcriptomic, and pharmacogenetic levels. These results support a fundamental role of the clock in pain pathophysiology. PLAIN LANGUAGE SUMMARY: Several headache diseases, such as migraine and cluster headache, have headaches that occur at the same time each day. We learned that the trigeminal ganglion, an important pain structure in several headache diseases, has a 24-hour cycle that might be related to this daily cycle of headaches. Our genetic analysis suggests that some medications may be more effective in treating migraine and cluster headache when taken at specific times of the day.

Characterization of the strain-rate-dependent mechanical response of single cell-cell junctions.

Cell-cell adhesions are often subjected to mechanical strains of different rates and magnitudes in normal tissue function. However, the rate-dependent mechanical behavior of individual cell-cell adhesions has not been fully characterized due to the lack of proper experimental techniques and therefore remains elusive. This is particularly true under large strain conditions, which may potentially lead to cell-cell adhesion dissociation and ultimately tissue fracture. In this study, we designed and fabricated a single-cell adhesion micro tensile tester (SCAµTT) using two-photon polymerization and performed displacement-controlled tensile tests of individual pairs of adherent epithelial cells with a mature cell-cell adhesion. Straining the cytoskeleton-cell adhesion complex system reveals a passive shear-thinning viscoelastic behavior and a rate-dependent active stress-relaxation mechanism mediated by cytoskeleton growth. Under low strain rates, stress relaxation mediated by the cytoskeleton can effectively relax junctional stress buildup and prevent adhesion bond rupture. Cadherin bond dissociation also exhibits rate-dependent strengthening, in which increased strain rate results in elevated stress levels at which cadherin bonds fail. This bond dissociation becomes a synchronized catastrophic event that leads to junction fracture at high strain rates. Even at high strain rates, a single cell-cell junction displays a remarkable tensile strength to sustain a strain as much as 200% before complete junction rupture. Collectively, the platform and the biophysical understandings in this study are expected to build a foundation for the mechanistic investigation of the adaptive viscoelasticity of the cell-cell junction.

Differentiation of salivary gland tumours using diffusion-weighted image-based virtual MR elastography: a pilot study.

OBJECTIVES: Differentiation among benign salivary gland tumours, Warthin tumours (WTs), and malignant salivary gland tumours is crucial to treatment planning and predicting patient prognosis. However, differentiation of those tumours using imaging findings remains difficult. This study evaluated the usefulness of elasticity determined from diffusion-weighted image (DWI)-based virtual MR elastography (MRE) compared with conventional magnetic resonance imaging (MRI) findings in differentiating the tumours. METHODS: This study included 17 benign salivary gland tumours, 6 WTs, and 11 malignant salivary gland tumours scanned on neck MRI. The long and short diameters, T1 and T2 signal intensities, tumour margins, apparent diffusion coefficient (ADC) values, and elasticity from DWI-based virtual MRE of the tumours were evaluated. The interobserver agreement in measuring tumour elasticity and the receiver operating characteristic (ROC) curves were also assessed. RESULTS: The long and short diameters and the T1 and T2 signal intensities showed no significant difference among the 3 tumour groups. Tumour margins and the mean ADC values showed significant differences among some tumour groups. The elasticity from virtual MRE showed significant differences among all 3 tumour groups and the interobserver agreement was excellent. The area under the ROC curves of the elasticity were higher than those of tumour margins and mean ADC values. CONCLUSION: Elasticity values based on DWI-based virtual MRE of benign salivary gland tumours, WTs, and malignant salivary gland tumours were significantly different. The elasticity of WTs was the highest and that of benign tumours was the lowest. The elasticity from DWI-based virtual MRE may aid in the differential diagnosis of salivary gland tumours.

The clock modulator Nobiletin mitigates astrogliosis-associated neuroinflammation and disease hallmarks in an Alzheimer's disease model.

Alzheimer's disease (AD) is a devastating neurodegenerative disorder, and there is a pressing need to identify disease-modifying factors and devise interventional strategies. The circadian clock, our intrinsic biological timer, orchestrates various cellular and physiological processes including gene expression, sleep, and neuroinflammation; conversely, circadian dysfunctions are closely associated with and/or contribute to AD hallmarks. We previously reported that the natural compound Nobiletin (NOB) is a clock-enhancing modulator that promotes physiological health and healthy aging. In the current study, we treated the double transgenic AD model mice, APP/PS1, with NOB-containing diets. NOB significantly alleviated ß-amyloid burden in both the hippocampus and the cortex, and exhibited a trend to improve cognitive function in these mice. While several systemic parameters for circadian wheel-running activity, sleep, and metabolism were unchanged, NOB treatment showed a marked effect on the expression of clock and clock-controlled AD gene expression in the cortex. In accordance, cortical proteomic profiling demonstrated circadian time-dependent restoration of the protein landscape in APP/PS1 mice treated with NOB. More importantly, we found a potent efficacy of NOB to inhibit proinflammatory cytokine gene expression and inflammasome formation in the cortex, and immunostaining further revealed a specific effect to diminish astrogliosis, but not microgliosis, by NOB in APP/PS1 mice. Together, these results underscore beneficial effects of a clock modulator to mitigate pathological and cognitive hallmarks of AD, and suggest a possible mechanism via suppressing astrogliosis-associated neuroinflammation.

Rho/ROCK mechanosensor in adipocyte stiffness and traction force generation.

It is increasingly recognized that interaction of adipose cells with extracellular mechanophysical milieus may play a role in regulating adipogenesis and differentiated adipocyte function and such interaction can be mediated by the mechanics of adipose cells. We measured the stiffness and traction force of adipose cells and examined the role of Rho/ROCK, the upstream effector of actin cytoskeletal contractility, in affecting these mechanical properties. Cellular Young's modulus obtained from atomic force microscopy (AFM) was significantly reduced by ROCK inhibitor (Y-27632) but elevated by Rho activator (CN01), for both preadipocytes and differentiated adipocytes. Immunofluorescent imaging suggested this could be attributed to the changes in Rho/ROCK-induced stressed actin filament formation. AFM also confirmed that differentiated adipocytes had higher stiffness than preadipocytes. On the other hand, traction force microscopy (TFM) revealed differentiated adipocytes exerted lower traction forces than preadipocytes. Traction forces of both preadipocytes and adipocytes were decreased by ROCK inhibition, but not significantly altered by Rho activation. Notably, an increasing trend of traction force with respect to cell spreading area was detected, and this trend was substantially amplified by Rho activation. Such traction force-cell area correlation was an order-of-magnitude smaller for differentiated adipocytes relative to preadipocytes, potentially due to disrupted force transmission through cytoskeleton-focal adhesion linkage by lipid droplets. Our work provides new data evidencing the Rho/ROCK control in adipose cell mechanics, laying the groundwork for adipocyte mechanotransduction studies on adipogenesis and adipose tissue remodeling.

Introducing a New Biomarker Named R2*-BOLD-MRI Parameter to Assess Treatment Response in Osteosarcoma.

BACKGROUND: While histologic response to neoadjuvant chemotherapy (NChT) is the major prognostic factor for osteosarcoma treatment, evaluating that response is difficult. PURPOSE: To evaluate the feasibility of the blood oxygen level-dependent (BOLD) technique to assess the response to NChT. STUDY TYPE: Prospective. POPULATION: Twelve patients with osteosarcoma undergoing NChT. FIELD STRENGTH/SEQUENCE: 3 T; T2*-weighted BOLD, dynamic contrast-enhanced (DCE) and diffusion-weighted (DW) (b values of 0, 400, and 1400 seconds/mm2 ) sequences. ASSESSMENT: Examination was performed before treatment (first), after each cycle of treatment (second and third). At each time point, spin dephasing rates (R2*) from BOLD magnetic resonance imaging (MRI), parameters from DCE-MRI (volume transfer constant [Ktrans ], reflux rate [kep ], volume fraction of the extravascular extracellular matrix [ve ], and blood plasma volume [vp ]), and the apparent diffusion coefficient (ADC) from DW-MRI were measured. STATISTICAL TESTS: Wilcoxon's signed rank test, Spearman's correlation coefficient (ρ) were used. A P-value of <0.05 was considered statistically significant. RESULTS: The difference and relative difference of the R2* values between the first/third MRIs in the extraosseous portion were statistically significant. Only the differences in the kep values between the first/second and between the first/third MRIs in the extraosseous portion were significant. The differences in the ADCs in the extraosseous and osseous portions were not statistically significant (P = 0.151, P = 0.733 each in extraosseous portion and P = 0.569, P = 0.129 each in osseous portion). The relative difference in R2* values in the extraosseous portion between the first/third MRI (ρ = 0.706) was significantly better correlated with the pathologic grade than those of kep and ADC over the same period (ρ = 0.286 and ρ = -0.091, respectively). DATA CONCLUSION: The R2* from the BOLD MRI technique could be a useful biomarker for evaluating treatment response in osteosarcoma treated with NchT. LEVEL OF EVIDENCE: 5 TECHNICAL EFFICACY: Stage 2.

Olsenella intestinalis sp. nov., isolated from cow feces.

A Gram-stain-negative, anaerobic, non-motile, rod-shaped bacterium, designated as BGYT1T, was isolated from the feces of a cow in Andong, Republic of Korea. It was studied using a polyphasic method to determine its taxonomic position. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain BGYT1T formed a lineage within the genus Olsenella and was most closely related to O. umbonate KCTC 15140T (98.2%). The complete genome sequence of strain BGYT1T was 2,476,083 bp long with a G + C content of 66.9 mol% and contained 1835 genes and 8 contigs. The N50 value was 604,117 bp. There were 50 tRNAs, 6 rRNAs (5S, 16S, 23S), 1778 CDSs and 2 BGCs and 1 tmRNA. The values for ANI (76.8%), AAI (67.3%), and dDDH (22.2%) compared to the closest related species were all below the threshold for bacterial species delineation. In addition, genes encoding the cell wall degrading enzymes such as chitinases, ß-1,3 glucanases, and proteases were also detected. The strain was able to grow at pH 6.0-8.0 (optimum, pH 7.0), in the presence of 0.5-1.5% NaCl (optimum, 0.5%, w/v) and at the temperature range of 35-40 °C (optimum, 35 °C). The predominant fatty acids were C16:0 DMA (20.2%), C16:0 (20.2%), C18:0 (10.5%) and C18:1 cis 9 (17.0%). The polar lipids consisted of an unidentified phospholipid, four unidentified glycolipids and three unidentified lipids. Based on its phenotypic analyses, phylogenetic and physiological characteristics, strain BGYT1T represented a novel species within the genus Olsenella, for which the name Olsenella intestinalis sp. nov. is proposed. The type strain is BGYT1T (= KCTC 25379T = GDMCC 1.3011T).

Comparison of Sensitivity Encoding (SENSE) and Compressed Sensing-SENSE for Contrast-Enhanced T1-Weighted Imaging in Patients With Crohn Disease Undergoing MR Enterography.

BACKGROUND. Long acquisition times for breath-hold contrast-enhanced (CE) T1-weighted imaging in MR enterography (MRE) protocols result in reduced image quality. OBJECTIVE. The purpose of this study was to compare CE T1-weighted imaging performed using sensitivity encoding (SENSE) and compressed sensing-SENSE (CS-SENSE) in terms of image quality and diagnostic performance for active inflammation in Crohn disease (CD). METHODS. This retrospective study included 41 patients (31 men, 10 women; mean age, 34 ± 12 [SD] years) who underwent MRE for known or suspected CD between June 2020 and September 2020. MRE was performed in one of two scanning rooms depending on scheduling availability. Per institutional protocol, in one room, the enteric phase was acquired using SENSE (acceleration factor, 3) and the portal phase was acquired using CS-SENSE (acceleration factor, 5); this order was reversed in the other room. Two radiologists independently assessed sequences for subjective image quality measures at the patient level and for active inflammation at the bowel-segment level. Mean image quality scores between readers were computed. Diagnostic performance for active inflammation was compared between SENSE and CS-SENSE using generalized estimating equations; a separate experienced radiologist reviewed the full MRE protocol to establish the reference standard. RESULTS. The mean acquisition time of CE T1-weighted imaging was 17.2 ± 1.1 seconds for SENSE versus 11.5 ± 0.8 seconds for CS-SENSE (p < .001). CS-SENSE scored significantly better than SENSE in overall image quality (4.2 ± 0.7 vs 3.7 ± 1.1; p = .02), motion artifacts (4.0 ± 0.8 vs 3.6 ± 1.2; p = .006), and aliasing artifacts (4.8 ± 0.4 vs 4.2 ± 0.6; p < .001). CS-SENSE scored significantly worse than SENSE in synthetic appearance (4.6 ± 0.5 vs 4.8 ± 0.4; p = .003). Contrast, sharpness, and blurring were not different between sequences (p > .05). For reader 1, CS-SENSE, compared with SENSE, showed a sensitivity of 86% versus 81% (p = .09), specificity of 88% versus 83% (p = .08), and accuracy of 87% versus 82% (p = .56). For reader 2, CS-SENSE, compared with SENSE, showed a sensitivity of 92% versus 79% (p = .006), specificity of 90% versus 98% (p = .16), and accuracy of 91% versus 86% (p = .002). CONCLUSION. Use of CS-SENSE for CE T1-weighted imaging in MRE protocols results in reduced scan times with reduced artifact and improved image quality. CLINICAL IMPACT. The benefits of CS-SENSE in MRE protocols may improve the diagnostic performance for active inflammation in CD.

Visible-Light Activation of a Dissolved Organic Matter-TiO2 Complex Mediated via Ligand-to-Metal Charge Transfer.

Given the widespread use of TiO2, its release into aquatic systems and complexation with dissolved organic matter (DOM) are highly possible, making it important to understand how such interactions affect photocatalytic activity under visible light. Here, we show that humic acid/TiO2 complexes (HA/TiO2) exhibit photoactivity (without significant electron-hole activation) under visible light through ligand-to-metal charge transfer (LMCT). The observed visible-light activities for pollutant removal and bacterial inactivation are primarily linked to the generation of H2O2via the conduction band. By systematically considering molecular-scale interactions between TiO2 and organic functional groups in HA, we find a key role of phenolic groups in visible-light absorption and H2O2 photogeneration. The photochemical formation of H2O2 in river waters spiked with TiO2 is notably elevated above naturally occurring H2O2 generated from background organic constituents due to LMCT contribution. Our findings suggest that H2O2 generation by HA/TiO2 is related to the quantity and functional group chemistry of DOM, which provides chemical insights into photocatalytic activity and potential ecotoxicity of TiO2 in environmental and engineered systems.

MST1 mediates the N-methyl-D-aspartate-induced excitotoxicity in mouse cortical neurons.

Excessive activation of the ionotropic N-methyl-D-aspartate (NMDA) receptor has been shown to cause abnormally high levels of Ca2+ influx, thereby leading to excitotoxic neuronal death. In this study, exposure of mouse primary cortical neurons to NMDA resulted in the cleavage and activation of mammalian sterile 20-like kinase-1 (MST1), both of which were mediated by calpain 1. In vitro cleavage assay data indicated that calpain 1 cleaves out the autoinhibitory domain of MST1 to generate an active form of the kinase. Furthermore, calpain 1 mediated the cleavage and activation of wild-type MST1, but not of MST1 (G339A). Intriguingly, NMDA/calpain-induced MST1 activation promoted the nuclear translocation of the kinase and the phosphorylation of histone H2B in mouse cortical neurons, leading to excitotoxicity. Thus, we propose a previously unrecognized mechanism of MST1 activation associated with NMDA-induced excitotoxic neuronal death.

TRAF6-mediated ubiquitination of MST1/STK4 attenuates the TLR4-NF-κB signaling pathway in macrophages.

Pattern-recognition receptors including Toll-like receptors (TLRs) recognize invading pathogens and trigger an immune response in mammals. Here we show that mammalian ste20-like kinase 1/serine/threonine kinase 4 (MST1/STK4) functions as a negative regulator of lipopolysaccharide (LPS)-induced activation of the TLR4-NF-κB signaling pathway associated with inflammation. Myeloid-specific genetic ablation of MST1/STK4 increased the susceptibility of mice to LPS-induced septic shock. Ablation of MST1/STK4 also enhanced NF-κB activation triggered by LPS in bone marrow-derived macrophages (BMDMs), leading to increased production of proinflammatory cytokines by these cells. Furthermore, MST1/STK4 inhibited TRAF6 autoubiquitination as well as TRAF6-mediated downstream signaling induced by LPS. In addition, we found that TRAF6 mediates the LPS-induced activation of MST1/STK4 by catalyzing its ubiquitination, resulting in negative feedback regulation by MST1/STK4 of the LPS-induced pathway leading to cytokine production in macrophages. Together, our findings suggest that MST1/STK4 functions as a negative modulator of the LPS-induced NF-κB signaling pathway during macrophage activation.

Effects of Donepezil Treatment on Brain Metabolites, Gut Microbiota, and Gut Metabolites in an Amyloid Beta-Induced Cognitive Impairment Mouse Pilot Model.

Accumulated clinical and biomedical evidence indicates that the gut microbiota and their metabolites affect brain function and behavior in various central nervous system disorders. This study was performed to investigate the changes in brain metabolites and composition of the fecal microbial community following injection of amyloid ß (Aß) and donepezil treatment of Aß-injected mice using metataxonomics and metabolomics. Aß treatment caused cognitive dysfunction, while donepezil resulted in the successful recovery of memory impairment. The Aß + donepezil group showed a significantly higher relative abundance of Verrucomicrobia than the Aß group. The relative abundance of 12 taxa, including Blautia and Akkermansia, differed significantly between the groups. The Aß + donepezil group had higher levels of oxalate, glycerol, xylose, and palmitoleate in feces and oxalate, pyroglutamic acid, hypoxanthine, and inosine in brain tissues than the Aß group. The levels of pyroglutamic acid, glutamic acid, and phenylalanine showed similar changes in vivo and in vitro using HT-22 cells. The major metabolic pathways in the brain tissues and gut microbiota affected by Aß or donepezil treatment of Aß-injected mice were related to amino acid pathways and sugar metabolism, respectively. These findings suggest that alterations in the gut microbiota might influence the induction and amelioration of Aß-induced cognitive dysfunction via the gut-brain axis. This study could provide basic data on the effects of Aß and donepezil on gut microbiota and metabolites in an Aß-induced cognitive impairment mouse model.

Variceal bleeding is aggravated by portal venous invasion of hepatocellular carcinoma: a matched nested case-control study.

BACKGROUND: We hypothesized that portal vein tumor thrombosis (PVTT) in hepatocellular carcinoma (HCC) increases portal pressure and causes esophageal varices and variceal bleedings. We examined the incidence of high-risk varices and variceal bleeding and determined the indications for variceal screening and prophylaxis. METHODS: This study included 1709 asymptomatic patients without any prior history of variceal hemorrhage or endoscopic prophylaxis who underwent upper endoscopy within 30 days before or after initial anti-HCC treatment. Of these patients, 206 had PVTT, and after 1:2 individual matching, 161 of them were matched with 309 patients without PVTT. High-risk varices were defined as large/medium varices or small varices with red-color signs and variceal bleeding. Bleeding rates from the varices were compared between matched pairs. Risk factors for variceal bleeding in the entire set of patients with PVTT were also explored. RESULTS: In the matched-pair analysis, the proportion of high-risk varices at screening (23.0% vs. 13.3%; P = 0.003) and the cumulative rate of variceal bleeding (4.5% vs. 0.4% at 1 year; P = 0.009) were significantly greater in the PVTT group. Prolonged prothrombin time, lower platelet count, presence of extrahepatic metastasis, and Vp4 PVTT were independent risk factors related to high-risk varices in the total set of 206 patients with PVTT (Adjusted odds ratios [95% CIs], 1.662 [1.151-2.401]; 0.985 [0.978-0.993]; 4.240 [1.783-10.084]; and 3.345 [1.457-7.680], respectively; Ps < 0.05). During a median follow-up of 43.2 months, 10 patients with PVTT experienced variceal bleeding episodes, 9 of whom (90%) had high-risk varices. Presence of high-risk varices and sorafenib use for HCC treatment were significant predictors of variceal bleeding in the complete set of patients with PVTT (Adjusted hazard ratios [95% CIs], 26.432 [3.230-216.289]; and 5.676 [1.273-25.300], respectively; Ps < 0.05). CONCLUSIONS: PVTT in HCC appears to increase the likelihood of high-risk varices and variceal bleeding. In HCC patients with PVTT, endoscopic prevention could be considered, at least in high-risk variceal carriers taking sorafenib.

Abbreviated Magnetic Resonance Imaging With Breath-Hold Three-Dimensional Magnetic Resonance Cholangiopancreatography: Assessment of Malignant Risk of Pancreatic Intraductal Papillary Mucinous Neoplasm.

BACKGROUND: For surveillance of pancreatic intraductal papillary mucinous neoplasms (IPMNs), magnetic resonance imaging (MRI)/magnetic resonance cholangiopancreatography (MRCP) is preferred over computed tomography, but the long acquisition time limits its use. PURPOSE: To investigate the diagnostic performance of abbreviated MRI with breath-hold (BH) three-dimensional MRCP (abbreviated MRI-BH) for malignant risk assessment of pancreatic IPMN. STUDY TYPE: Retrospective. POPULATION: Two hundred and thirty-five patients with IPMNs (M:F = 115:120; mean age ± SD, 66 ± 9 years; typical imaging features with ≥2-year stability [N = 172] and histopathologically confirmed [N = 63]). FIELD STRENGTH/SEQUENCE: 3.0 T/ abbreviated MRI-BH (single-shot fast spin-echo, T1W fat-suppressed gradient-echo sequence, and BH-3D-MRCP). ASSESSMENT: Abbreviated MRI-BH was reviewed by three reviewers, and its diagnostic performance was assessed using the predetermined scoring system. The diagnostic performance for the mural nodule detection was assessed. Additionally, diagnostic performance of abbreviated MRI was compared with that of full-sequence MRI. STATISTICAL TESTS: Area under the receiver operating characteristic curve (AUC) with z-test, and linear-weighted kappa values. RESULTS: Thirty-five patients had malignant IPMN. At a cut-off score ≥3, AUCs of abbreviated MRI-BH for detecting malignant IPMN were 0.959 for reviewer 1, 0.962 for reviewer 2, and 0.956 for reviewer 3. The sensitivity of reviewers 1, 2, and 3 was 97.1% for all, and the specificity was 85.5%, 86.0%, and 85.0%, respectively. Regarding mural nodule detection (N = 22), abbreviated MRI-BH demonstrated a sensitivity of 95.5% and a specificity of 88.3% for reviewer 1, a sensitivity of 86.4% and a specificity of 92.0% for reviewer 2, and a sensitivity of 86.4% and a specificity of 89.2% for reviewer 3. There were no significant differences between AUC of abbreviated MRI-BH and that of full-sequence MRI in the three reviewers (P > 0.05). DATA CONCLUSION: Abbreviated MRI-BH showed good diagnostic performance for detecting malignant IPMNs by using a predetermined scoring system. LEVEL OF EVIDENCE: 3 TECHNICAL EFFICACY STAGE: 2.

Low conductivity on electrical properties tomography demonstrates unique tumor habitats indicating progression in glioblastoma.

OBJECTIVES: Tissue conductivity measurements made with electrical properties tomography (EPT) can be used to define temporal changes in tissue habitats on longitudinal multiparametric MRI. We aimed to demonstrate the added insights for identifying tumor habitats obtained by including EPT with diffusion- and perfusion-weighted MRI, and to evaluate the use of these tumor habitats for determining tumor treatment response in post-treatment glioblastoma. METHODS: Tumor habitats were developed from EPT, diffusion-weighted, and perfusion-weighted MRI in 60 patients with glioblastoma who underwent concurrent chemoradiotherapy. Voxels from EPT, apparent diffusion coefficient (ADC), and cerebral blood volume (CBV) maps were clustered into habitats, and each habitat was serially examined to assess its temporal change. The usefulness of temporal changes in tumor habitats for diagnosing tumor progression and treatment-related change was investigated using logistic regression. The performance of significant predictors was measured using the area under the curve (AUC) from receiver-operating-characteristics analysis with 1000-fold bootstrapping. RESULTS: Five tumor habitats were identified, and of these, the hypervascular cellular habitat (odds ratio [OR] 5.45; 95% CI, 1.75-31.42; p = .02), hypovascular low conductivity habitat (OR 2.00; 95% CI, 1.45-3.05; p < .001), and hypovascular intermediate habitat (OR 1.57; 95% CI, 1.18-2.30; p = .006) were predictive of tumor progression. Low EPT and low CBV reflected a unique hypovascular low conductivity habitat that showed the highest diagnostic performance (AUC 0.86; 95% CI, 0.76-0.96). The combined habitats showed high performance (AUC 0.90; 95% CI, 0.82-0.98) in the differentiation of tumor progression from treatment-related change. CONCLUSION: EPT reveals low conductivity habitats that can improve the diagnosis of tumor progression in post-treatment glioblastoma. KEY POINTS: • Electrical properties tomography (EPT) demonstrated lower conductivity in tumor progression than in treatment-related change. • EPT allowed identification of a unique hypovascular low conductivity habitat when combined with cerebral blood volume mapping. • Tumor habitats with a hypovascular low conductivity habitat, hypervascular cellular habitat, and hypovascular intermediate habitat yielded high diagnostic performance for diagnosing tumor progression.

Comparisons between image quality and diagnostic performance of 2D- and breath-hold 3D magnetic resonance cholangiopancreatography at 3T.

OBJECTIVES: To compare the image quality and diagnostic performance of 2D MRCP to those of breath-hold 3D MRCP using compressed sensing (CS-MRCP) and gradient and spin-echo (GRASE-MRCP) at 3T. METHODS: From January to November 2018, patients who underwent pancreatobiliary MRI including 2D MRCP and two breath-hold 3D MRCP using CS and GRASE at 3T were included. Three radiologists independently evaluated image quality, motion artifact, and pancreatic cyst conspicuity. Diagnostic performance was assessed for bile duct anatomic variation, bile duct, and pancreatic diseases using a composite algorithm as reference standards. Pancreatic lesion detectability and conspicuity were evaluated using JAFROC and generalized estimating equation analysis. RESULTS: One hundred patients (male = 50) were included. Bile duct anatomic variation, bile duct and pancreatic diseases were present in respectively 31, 15, and 79 patients. Breath-hold 3D MRCP provided better image quality than 2D MRCP (3.5 ± 0.6 in 2D MRCP; 4.0 ± 0.7 in GRASE-MRCP and 3.9 ± 0.8 in CS-MRCP, p < 0.001 for both). There was no difference in motion artifact between 2D and breath-hold 3D MRCP (p = 0.1). Breath-hold 3D CS-MRCP provided better pancreatic cyst conspicuity than 2D MRCP (2.7 [95% CI: 2.5-3.0] vs. 2.3 [95% CI: 2.1-2.5], p = 0.001). There were no significant differences between the diagnostic performance of the three sequences in the detection of bile duct anatomic variation or pancreatic lesions (p > 0.05). CONCLUSION: Breath-hold 3D MRCP with GRASE or CS can provide better image quality than 2D MRCP in a comparable scan time. KEY POINTS: • Breath-hold 3D MRCP using compressed sensing (CS) or gradient and spin-echo (GRASE) provided a better image quality with less image blurring than 2D MRCP. • There were no significant differences between 2D MRCP and breath-hold 3D MRCP in either motion artifact or the number of non-diagnostic exams. • There were no significant differences between 2D MRCP and either type of breath-hold 3D MRCP in the diagnosis of bile duct anatomic variation or detection of pancreatic lesions.