Liver-specific Lxr inhibition represses reverse cholesterol transport in cholesterol-fed mice.

BACKGROUND AND AIMS: High density lipoprotein (HDL) exerts an anti-atherosclerotic effect via reverse cholesterol transport (RCT). Several phases of RCT are transcriptionally controlled by Liver X receptors (Lxrs). Although macrophage Lxrs reportedly promote RCT, it is still uncertain whether hepatic Lxrs affect RCT in vivo. METHODS: To inhibit Lxr-dependent pathways in mouse livers, we performed hepatic overexpression of sulfotransferase family cytosolic 2B member 1 (Sult2b1) using adenoviral vector (Ad-Sult2b1). Ad-Sult2b1 or the control virus was intravenously injected into wild type mice and Lxrα/ß double knockout mice, under a normal or high-cholesterol diet. A macrophage RCT assay and an HDL kinetic study were performed. RESULTS: Hepatic Sult2b1 overexpression resulted in reduced expression of Lxr-target genes - ATP-binding cassette transporter G5/G8, cholesterol 7α hydroxylase and Lxrα itself - respectively reducing or increasing cholesterol levels in HDL and apolipoprotein B-containing lipoproteins (apoB-L). A macrophage RCT assay revealed that Sult2b1 overexpression inhibited fecal excretion of macrophage-derived 3H-cholesterol only under a high-cholesterol diet. In an HDL kinetic study, Ad-Sult2b1 promoted catabolism/hepatic uptake of HDL-derived cholesterol, thereby reducing fecal excretion. Finally, in Lxrα/ß double knockout mice, hepatic Sult2b1 overexpression increased apoB-L levels, but there were no differences in HDL levels or RCT compared to the control, indicating that Sult2b1-mediated effects on HDL/RCT and apoB-L were distinct: the former was Lxr-dependent, but not the latter. CONCLUSIONS: Hepatic Lxr inhibition negatively regulates circulating HDL levels and RCT by reducing Lxr-target gene expression.

A New Case of Abetalipoproteinemia Caused by Novel Compound Heterozygote Mutations in the MTTP Gene without Fat or Vitamin Malabsorption.

Abetalipoproteinemia (ABL) is a rare disease characterized by extremely low apolipoprotein B (apoB)-containing lipoprotein levels, dietary fat, and fat-soluble vitamin malabsorption, leading to gastrointestinal, neuromuscular, and ophthalmological symptoms. We herein report a case of ABL with novel compound heterozygous mutations in the microsomal triglyceride transfer protein gene (c.1686_1687del [p.Ser563TyrfsTer10] and c.1862T>C [p.Ile621Thr]), identified via panel sequencing. Although the patient had extremely reduced low-density lipoprotein cholesterol levels and a fatty liver, he did not exhibit other typical complications. Furthermore, unlike typical ABL, this patient had a preserved apoB-48 secretion and increased concentrations of high-density lipoprotein cholesterol, which may account for the normal serum fat-soluble vitamin levels.

Graphical user interface-based convolutional neural network models for detecting nasopalatine duct cysts using panoramic radiography.

Nasopalatine duct cysts are difficult to detect on panoramic radiographs due to obstructive shadows and are often overlooked. Therefore, sensitive detection using panoramic radiography is clinically important. This study aimed to create a trained model to detect nasopalatine duct cysts from panoramic radiographs in a graphical user interface-based environment. This study was conducted on panoramic radiographs and CT images of 115 patients with nasopalatine duct cysts. As controls, 230 age- and sex-matched patients without cysts were selected from the same database. The 345 pre-processed panoramic radiographs were divided into 216 training data sets, 54 validation data sets, and 75 test data sets. Deep learning was performed for 400 epochs using pretrained-LeNet and pretrained-VGG16 as the convolutional neural networks to classify the cysts. The deep learning system's accuracy, sensitivity, and specificity using LeNet and VGG16 were calculated. LeNet and VGG16 showed an accuracy rate of 85.3% and 88.0%, respectively. A simple deep learning method using a graphical user interface-based Windows machine was able to create a trained model to detect nasopalatine duct cysts from panoramic radiographs, and may be used to prevent such cysts being overlooked during imaging.

Protection of Liver Functions and Improvement of Kidney Functions by Twelve Weeks Consumption of Cuban Policosanol (Raydel®) with a Decrease of Glycated Hemoglobin and Blood Pressure from a Randomized, Placebo-Controlled, and Double-Blinded Study with Healthy and Middle-Aged Japanese Participants.

Policosanol consumption has been associated with treating blood pressure and dyslipidemia by increasing the level of high-density lipoproteins-cholesterol (HDL-C) and HDL functionality. Although policosanol supplementation also ameliorated liver function in animal models, it has not been reported in a human clinical study, particularly with a 20 mg doage of policosanol. In the current study, twelve-week consumption of Cuban policosanol (Raydel®) significantly enhanced the hepatic functions, showing remarkable decreases in hepatic enzymes, blood urea nitrogen, and glycated hemoglobin. From the human trial with Japanese participants, the policosanol group (n = 26, male 13/female 13) showed a remarkable decrease in alanine aminotransferase (ALT) and aspartate aminotransferase (AST) from baseline up to 21% (p = 0.041) and 8.7% (p = 0.017), respectively. In contrast, the placebo group (n = 26, male 13/female 13) showed almost no change or slight elevation. The policosanol group showed a 16% decrease in γ-glutamyl transferase (γ-GTP) at week 12 from the baseline (p = 0.015), while the placebo group showed a 1.2% increase. The policosanol group exhibited significantly lower serum alkaline phosphatase (ALP) levels at week 8 (p = 0.012), week 12 (p = 0.012), and after 4-weeks (p = 0.006) compared to those of the placebo group. After 12 weeks of policosanol consumption, the ferric ion reduction ability and paraoxonase of serum were elevated by 37% (p < 0.001) and 29% (p = 0.004) higher than week 0, while placebo consumption showed no notable changes. Interestingly, glycated hemoglobin (HbA1c) in serum was lowered significantly in the policosanol group 4 weeks after consumption, which was approximately 2.1% (p = 0.004) lower than the placebo group. In addition, blood urea nitrogen (BUN) and uric acid levels were significantly lower in the policosanol group after 4 weeks: 14% lower (p = 0.002) and 4% lower (p = 0.048) than those of the placebo group, respectively. Repeated measures of ANOVA showed that the policosanol group had remarkable decreases in AST (p = 0.041), ALT (p = 0.008), γ-GTP (p = 0.016), ALP (p = 0.003), HbA1c (p = 0.010), BUN (p = 0.030), and SBP (p = 0.011) from the changes in the placebo group in point of time and group interaction. In conclusion, 12 weeks of 20 mg consumption of policosanol significantly enhanced hepatic protection by lowering the serum AST, ALT, ALP, and γ-GTP via a decrease in glycated hemoglobin, uric acid, and BUN with an elevation of serum antioxidant abilities. These results suggest that improvements in blood pressure by consumption of 20 mg of policosanol (Raydel®) were accompanied by protection of liver function and enhanced kidney function.

Beneficial Effect of Cuban Policosanol on Blood Pressure and Serum Lipoproteins Accompanied with Lowered Glycated Hemoglobin and Enhanced High-Density Lipoprotein Functionalities in a Randomized, Placebo-Controlled, and Double-Blinded Trial with Healthy Japanese.

This study evaluated the efficacy and safety of 20 mg of Cuban policosanol in blood pressure (BP) and lipid/lipoprotein parameters of healthy Japanese subjects via a placebo-controlled, randomized, and double-blinded human trial. After 12 weeks of consumption, the policosanol group showed significantly lower BP, glycated hemoglobin (HbA1c), and blood urea nitrogen (BUN) levels. The policosanol group also showed lower aspartate aminotransferase (AST), alanine aminotransferase (ALT), and γ-glutamyl transferase (γ-GTP) levels at week 12 than those at week 0: A decrease of up to 9% (p < 0.05), 17% (p < 0.05), and 15% (p < 0.05) was observed, respectively. The policosanol group showed significantly higher HDL-C level and HDL-C/TC (%), approximately 9.5% (p < 0.001) and 7.2% (p = 0.003), respectively, than the placebo group and a difference in the point of time and group interaction (p < 0.001). In lipoprotein analysis, the policosanol group showed a decrease in oxidation and glycation extent in VLDL and LDL with an improvement of particle shape and morphology after 12 weeks. HDL from the policosanol group showed in vitro stronger antioxidant and in vivo anti-inflammatory abilities. In conclusion, 12 weeks of Cuban policosanolconsumption in Japanese subjects showed significant improvement in blood pressure, lipid profiles, hepatic functions, and HbA1c with enhancement of HDL functionalities.

The Validity of Ultra-Short-Term Heart Rate Variability during Cycling Exercise.

Ultra-short-term heart rate variability (HRV) has been validated in the resting state, but its validity during exercise is unclear. This study aimed to examine the validity in ultra-short-term HRV during exercise considering the different exercise intensities. HRVs of twenty-nine healthy adults were measured during incremental cycle exercise tests. HRV parameters (Time-, frequency-domain and non-linear) corresponding to each of the 20% (low), 50% (moderate), and 80% (high) peak oxygen uptakes were compared between the different time segments of HRV analysis (180 s (sec) segment vs. 30, 60, 90, and 120-sec segments). Overall, the differences (bias) between ultra-short-term HRVs increased as the time segment became shorter. In moderate- and high-intensity exercises, the differences in ultra-short-term HRV were more significant than in low intensity exercise. Thus, we discovered that the validity of ultra-short-term HRV differed with the duration of the time segment and exercise intensities. However, the ultra-short-term HRV is feasible in the cycling exercise, and we determined some optimal time duration for HRV analysis for across exercise intensities during the incremental cycling exercise.

Dipeptidyl Peptidase-4 Inhibitor Sitagliptin Phosphate Accelerates Cellular Cholesterol Efflux in THP-1 Cells.

Cholesterol efflux is a major atheroprotective function of high-density lipoproteins (HDLs) which removes cholesterol from the foam cells of lipid-rich plaques in Type 2 diabetes. The dipeptidyl peptidase-4 (DPP-4) inhibitor sitagliptin phosphate increases plasma glucagon-like peptide-1 (GLP-1) concentrations and is used to treat Type 2 diabetes. GLP-1 plays an important role in regulating insulin secretion and expression via the GLP-1 receptor (GLP-1R), which is expressed in pancreatic islets as well as freshly isolated human monocytes and THP-1 cells. Here, we identified a direct role of GLP-1 and DPP-4 inhibition in HDL function. Cholesterol efflux was measured in cultivated phorbol 12-myristate 13-acetate-treated THP-1 cells radiolabeled with 3H-cholesterol and stimulated with liver X receptor/retinoid X receptor agonists. Contrary to vildagliptin, sitagliptin phosphate together with GLP-1 significantly (p < 0.01) elevated apolipoprotein (apo)A1-mediated cholesterol efflux in a dose-dependent manner. The sitagliptin-induced increase in cholesterol efflux did not occur in the absence of GLP-1. In contrast, adenosine triphosphate-binding cassette transporter A1 (ABCA1) mRNA and protein expressions in the whole cell fraction were not changed by sitagliptin in the presence of GLP-1, although sitagliptin treatment significantly increased ABCA1 protein expression in the membrane fraction. Furthermore, the sitagliptin-induced, elevated efflux in the presence of GLP-1 was significantly decreased by a GLP-1R antagonist, an effect that was not observed with a protein kinase A inhibitor. To our knowledge, the present study reports for the first time that sitagliptin elevates cholesterol efflux in cultivated macrophages and may exert anti-atherosclerotic actions that are independent of improvements in glucose metabolism. Our results suggest that sitagliptin enhances HDL function by inducing a de novo HDL synthesis via cholesterol efflux.

Two Cases of Acquired High-Density Lipoprotein Deficiency with Immunoglobulin G4-Related Lecithin-Cholesterol Acyltransferase Autoantibody.

Lecithin-cholesterol acyltransferase (LCAT) plays a significant role in the progression from premature to mature high-density lipoprotein (HDL) in circulation. Consequently, primary or secondary LCAT deletion or reduction naturally results in low serum HDL cholesterol levels. Recently, rare cases of acquired HDL deficiency with LCAT autoantibodies have been reported, mainly from Japan, where LCAT autoantibodies of immunoglobulin G (IgG) caused the HDL deficiency. Here to our knowledge, we report for the first time two cases of acquired HDL deficiency caused by IgG4 linked LCAT autoantibodies with or without a high serum IgG4 level. Furthermore, these cases can extend to a new concept of "IgG4 autoimmune disease" from the viewpoint of verifying the serum autoantibody and/or renal histopathology.

Cuban policosanol improves high-density lipoprotein cholesterol efflux capacity in healthy Japanese subjects.

Policosanol supplementation has been reported to increase high-density lipoprotein (HDL)-cholesterol (HDL-C). However, the association between Cuban policosanol supplementation and HDL cholesterol efflux capacity (CEC), an important function of HDL, remains unclear. We performed a lipoprotein analysis investigating 32 Japanese healthy participants (placebo, n = 17 or policosanol supplementation for 12 weeks, n = 15) from a randomized Cuban policosanol clinical trial. First, HDL CEC and HDL-related factors were measured before and after policosanol supplementation. Then, through electron microscopy after ultracentrifugation and high-performance liquid chromatography, HDL morphology and subclass were analyzed, respectively. Finally, the effects of policosanol supplementation regarding HDL function, HDL-related factors, and HDL morphology/component were examined. Cuban policosanol considerably increased the HDL CEC and HDL-C and apolipoprotein A-I (ApoA-I) levels. Furthermore, policosanol supplementation led to larger HDL particles, increased cholesterol content in larger HDL particles, and reduced triglyceride content in smaller HDL particles. In participants with high baseline HDL-C levels, the policosanol effects for HDL CEC are observed. HDL CEC fluctuation induced by policosanol was highly associated with HDL-C and ApoA-I changes. In conclusion, for the first time, we demonstrated that policosanol supplementation increased the HDL CEC in healthy participants.

Quantitative assessment of the mandibular condyle in patients with diabetes mellitus using diffusion-weighted magnetic resonance imaging.

OBJECTIVE: The purpose of this study was to quantitatively assess the mandibular condyle in patients with diabetes mellitus (DM) using the apparent diffusion coefficient (ADC) on diffusion-weighted magnetic resonance imaging (DWI). STUDY DESIGN: 102 patients with DM and temporomandibular joint (TMJ) pain who underwent magnetic resonance imaging (MRI) examination of the TMJs at our hospital between August 2006 and March 2020 were included in this study. 112 patients with temporomandibular disorders (TMD) who underwent MRI examination at our hospital between April 2019 and March 2020 were included as controls. The MRI findings were compared between the two groups. RESULTS: The mean ADC values of the mandibular condyle in patients with DM were significantly greater than the controls (P < 0.01). Receiver operating characteristic (ROC) curve analysis revealed a cutoff of 0.98 for the ADC values of the mandibular condyle in patients with DM. CONCLUSION: This study found that the ADC on DWI could be used for the quantitative assessment of the mandibular condyle in patients with DM. DWI might serve as a new and noninvasive method to assess the presence of DM.

The diagnostic utility of size and apparent diffusion coefficient values for cervical lymph nodes in patients with osteomyelitis of the jaw bone.

PURPOSE: The purpose of this study was to (1) clarify the size and apparent diffusion coefficient (ADC) value of lymph nodes (LN) in each state in their quantitative evaluation diffusion-weighted imaging, and (2) to determine the diagnostic utility of size and ADC values in the quantitative evaluation of LNs using diffusion-weighted imaging. METHODS: This was a retrospective cohort study at our hospital conducted between April 2017 and March 2019. A total of 50 patients (20 men, 30 women) with 118 LNs, aged 34-90 years (mean age 61.18 years), undergoing magnetic resonance imaging examination were included in the study. The predictor variable was disease status. The primary outcome variable was the mean size and ADC values of the LNs. The other variables were age and sex. Data were analyzed using a Kruskal-Wallis test, and hoc Mann-Whitney tests with Bonferroni adjustment and a receiver operating characteristic (ROC) curve. P < 0.05 was considered to indicate statistical significance. RESULTS: We analyzed the records of 50 patients (118 LNs) with and without osteomyelitis. Of these, 21 had acute osteomyelitis, and 16 had chronic osteomyelitis. The size and ADC values of LNs in the osteomyelitis group were significantly greater and higher, respectively, than those in the non-myelitis group (P < 0.01). ROC analysis revealed a cutoff short-axis size of 4.42 and 4.04 mm for lymphadenopathy caused by osteomyelitis, corresponding to levels IB and level II, respectively. Moreover, the ADC cutoff values for the same were 0.85 and 0.86, respectively. CONCLUSION: The results suggest that size and ADC values are useful parameters for the quantitative evaluation of lymphadenopathy caused by osteomyelitis.

Quantitative assessment of the mandibular bone marrow of diabetes mellitus patients using diffusion-weighted magnetic resonance imaging.

PURPOSE: The purpose of this study was to assess quantitatively the mandibular bone marrow of patients with and without diabetes mellitus (DM) using the apparent diffusion coefficient (ADC) values on diffusion-weighted imaging (DWI). METHODS: 65 DM patients (28 men, 37 women, 29-84 years of age, mean age 55.7 ± 15.7 years) and age-, sex- and periodontitis stage-matched 65 non-DM patients who had underwent MRI between April 2006 and March 2018 were included in this study. The ADC was calculated using the ADC visualization tool implemented in a dedicated off-line workstation. The regions of interest (ROI) were manually placed on the ADC map on which the mandibular bone marrow from the lower first molar to the lower second molar was observed in patients with and without DM. Statistical analysis was performed using the Mann-Whitney U test and receiver operating characteristic (ROC) curve analysis. P values < 0.05 were considered statistically significant. RESULTS: The mean ADC values of the mandibular bone marrow of patients with and without DM were 1.18 ± 0.21 × 10-3 mm2/s and 0.83 ± 0.14 × 10-3 mm2/s, respectively. The ADC values of DM patients were significantly higher than those of patients without DM. CONCLUSION: The ADC values allowed the quantitative evaluation of the mandibular bone marrow of DM patients. DWI might serve as a new and noninvasive method to assess the presence of DM.

Atorvastatin Reduces Circulating S100A12 Levels in Patients with Carotid Atherosclerotic Plaques - A Link with Plaque Inflammation.

AIMS: Inflammation is involved in various processes of atherosclerosis development. Serum C-reactive protein (CRP) levels, a predictor for cardiovascular risk, are reportedly reduced by statins. However, several studies have demonstrated that CRP is a bystander during atherogenesis. While S100A12 has been focused on as an inflammatory molecule, it remains unclear whether statins affect circulating S100A12 levels. Here, we investigated whether atorvastatin treatment affected S100A12 and which biomarkers were correlated with changes in arterial inflammation. METHODS: We performed a prospective, randomized open-labeled trial on whether atorvastatin affected arterial (carotid and thoracic aorta) inflammation using 18fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG-PET/CT) and inflammatory markers. Thirty-one statin-naïve patients with carotid atherosclerotic plaques were randomized to either a group receiving dietary management (n=15) or one receiving atorvastatin (10mg/day, n=16) for 12weeks. 18F-FDG-PET/CT and flow-mediated vasodilation (FMD) were performed, the latter to evaluate endothelial function. RESULTS: Atorvastatin, but not the diet-only treatment, significantly reduced LDL-cholesterol (LDL-C, -43%), serum CRP (-37%) and S100A12 levels (-28%) and improved FMD (+38%). 18F-FDG-PET/CT demonstrated that atorvastatin, but not the diet-only treatment, significantly reduced accumulation of 18F-FDG in the carotid artery and thoracic aorta. A multivariate analysis revealed that reduction in CRP, S100A12, LDL-C, oxidized-LDL, and increase in FMD were significantly associated with reduced arterial inflammation in the thoracic aorta, but not in the carotid artery. CONCLUSIONS: Atorvastatin treatment reduced S100A12/CRP levels, and the changes in these circulating markers mirrored the improvement in arterial inflammation. Our observations suggest that S100A12 may be an emerging therapeutic target for atherosclerosis.

The ratio of heart rate to heart rate variability reflects sympathetic activity during incremental cycling exercise.

A low-frequency to a high-frequency component ratio (LF/HF) in heart rate variability (HRV) may not accurately reflect sympathetic nervous activity during exercise. Thus, a valid HRV-based index of sympathetic nervous activity is needed. Therefore, the heart rate to LF ratio (Heart rate/LF) was evaluated as sympathetic nervous activity index which is reflected by catecholamine levels during incremental exercise. In this study, 15 healthy adults performed an incremental exercise test using a cycle ergometer. HRV was derived from electrocardiography and HRV components related to the autonomic nervous system were obtained using frequency analysis. Heart rate/LF was calculated using the heart rate and LF component produced by HRV analysis. Catecholamine, blood lactate levels and respiratory gas were also measured throughout the exercise test. While LF/HF did not increase with increasing exercise intensity, Heart rate/LF non-linearly increased during the incremental exercise test, as did noradrenaline and blood lactate. Interestingly, Heart rate/LF values were positively correlated with noradrenaline (ρ = 0.788, p < 0.05) and blood lactate (ρ = 0.802, p < 0.05) levels and carbon dioxide production (ρ = 0.903, p < 0.05) from at rest through the exercise stages. Heart rate/LF reflects sympathetic nervous activity and metabolic responses during incremental cycling exercise and has potential as an HRV index of sympathetic nervous activity during exercise.Trial registration: UMIN Japan identifier: UMIN000039639.

Highly Efficient Multi-Step Oxidation Bioanode Using Microfluidic Channels.

With the rapid decline of fossil fuels, various types of biofuel cells (BFCs) are being developed as an alternative energy source. BFCs based on multi-enzyme cascade reactions are utilized to extract more electrons from substrates. Thus, more power density is obtained from a single molucule of substrate. In the present study, a bioanode that could extract six electrons from a single molecule of L-proline via a three-enzyme cascade reaction was developed and investigated for its possible use in BFCs. These enzymes were immobilized on the electrode to ensure highly efficient electron transfer. Then, oriented immobilization of enzymes was achieved using two types of self-assembled monolayers (SAMs). In addition, a microfluidic system was incorporated to achieve efficient electron transfer. The microfluidic system, in which the electrodes were arranged in a tooth-shaped comb, allowed for substrates to be supplied continuously to the cascade, which resulted in smooth electron transfer. Finally, we developed a high-performance bioanode which resulted in the accumulation of higher current density compared to that of a gold disc electrode (205.8 µA cm-2: approximately 187 times higher). This presents an opportunity for using the bioanode to develop high-performance BFCs in the future.

Specificity of tuberculin skin test improved by BCG immunization schedule change in Japan.

INTRODUCTION: Tuberculin skin test (TST) has been used to diagnose tuberculosis (TB) and latent tuberculosis infection (LTBI). However, in Bacillus Calmette-Guérin (BCG) vaccinated patients, TST tends to produce false-positive results. According to the previous vaccination schedule, Japanese people were mandated to receive up to three doses of BCG-vaccine. The vaccination schedule was changed in 2003 and as per the new schedule, only infants are administered a dose of BCG vaccine. Our hypothesis is that this change can lead to a reduction in the cross-reaction to TST. METHODS: We evaluated the TST results obtained from 1097 recruits from six defense camps and 667 recruits from an air base. These TST data were divided into two groups according to the date of birth: a new group and an old group according to the BCG immunization schedule. We then analyzed positive and negative reaction of TST and erythema sizes. RESULTS: We confirmed that the change in BCG-vaccination schedule significantly decreased TST false-positive reaction (Pmeta = 1.4 × 10-18; risk ratio = 0.83; 95% confidence interval: 0.80-0.87) and erythema size (Pmeta = 1.1 × 10-4; mean difference = 6.6 mm; 95% confidence interval: 3.2 mm-9.9 mm). CONCLUSIONS: We showed the reduction in BCG cross-reaction to TST, in the new BCG vaccination schedule group, compared to the old group, we also have extracted information on the improvement in the specificity of TST for LTBI and TB diagnosis, which resulted from BCG schedule change.