Apple Pomace Extract Improves MK-801-Induced Memory Impairment in Mice.

Alzheimer's disease (AD) is a neurodegenerative disease that involves progressive cognitive decline accompanied by synaptic degeneration and impaired neurotransmission. Recent studies revealed that apple pomace, a waste byproduct of the apple processing industry, has beneficial health properties, but its potential to prevent and treat AD has not been determined. Herein, we examined the effects of apple pomace extract on N-methyl-D-aspartate receptor antagonist MK-801-induced memory impairment in mice. Repeated treatment with apple pomace extract for 7 days reversed the MK-801-induced impairment of associative memory and recognition memory. RNA sequencing revealed that repeated treatment with apple pomace extract altered the gene expression profile in the hippocampus of mice. Real-time PCR showed that apple pomace extract induced upregulation of the mRNA expression for Zfp125 and Gstp1. Furthermore, gene sets related to synapse and neurotransmission were upregulated by apple pomace extract. These findings indicate that apple pomace extract may be useful for the prevention and treatment of AD.

Mechanism of Negative Thermal Expansion in Monoclinic Cu2P2O7 from First Principles.

Negative thermal expansion (NTE) materials generally have high-symmetry space groups, large average atomic volumes, and corner-sharing octahedral and tetrahedral coordination structures. By contrast, monoclinic α-Cu2P2O7, which has a small average atomic volume and edge-sharing structure, has been reported to exhibit NTE, the detailed mechanism of which is unclear. In this study, we investigate the A2B2O7 polymorphs and analyze the NTE behavior of α-Cu2P2O7 using first-principles lattice-dynamics calculations. From the polymorphism investigation in 20 A2B2O7 compounds using 6 representative crystal structures, small A and B cationic radii are found to stabilize the α-Cu2P2O7-type structure. We then analyze the NTE behavior of α-Cu2P2O7 using quasi-harmonic approximation. Our calculated thermal expansion coefficients and anisotropic atomic displacement parameters were in good agreement with those of the experimental reports at low temperatures. From the mode-Grüneisen parameter distribution plotted over the entire first-Brillouin zone, we found that the phonon contributing most significantly to NTE emerges not into the special points but between them. In this phonon mode, the O connecting two PO4 tetrahedra rotates, and the Cu and O vibrate perpendicular to the bottom of the CuO5 pyramidal unit, which folds the ac lattice plane. This vibration behavior can explain the experimentally reported anisotropic NTE behavior of α-Cu2P2O7. Our results demonstrate that the most negative mode-Grüneisen parameter contributing to NTE behavior is not always located on high-symmetry special points, indicating the importance of lattice vibration analyses for the entire first-Brillouin zone.

A polysaccharide from Inula japonica showing in vivo antitumor activity by interacting with TLR-4, PD-1, and VEGF.

Polysaccharides, an important class of carbohydrate polymers, are considered as one of the sources of drug molecules. To discover bioactive polysaccharides as potential agents against cancer, a homogeneous polysaccharide (IJP70-1) has been purified from the flowers of Inula japonica, which is a traditional medicinal plant used for various medical indications. IJP70-1 with a molecular weight of 1.019 × 105 Da was mainly composed of →5)-α-l-Araf-(1→, →2,5)-α-l-Araf-(1→, →3,5)-α-l-Araf-(1→, →2,3,5)-α-l-Araf-(1→, →6)-α-d-Glcp-(1→, →3,6)-α-d-Galp-(1→, and t-α-l-Araf. Apart from the characteristics and structure elucidated by various techniques, the in vivo antitumor activity of IJP70-1 was assayed using zebrafish models. In the subsequent mechanism investigation, it was found that the in vivo antitumor activity of IJP70-1 was not cytotoxic mechanism caused, but related to the activation of the immune system and inhibition of angiogenesis by interacting with the proteins toll-like receptor-4 (TLR-4), programmed death receptor-1 (PD-1), and vascular endothelial growth factor (VEGF). The chemical and biological studies have shown that the homogeneous polysaccharide IJP70-1 has the potential to be developed into an anticancer agent.

A Histogram Analysis of the Pixel Grayscale (Luminous Intensity) of B-Mode Ultrasound Images of the Subcutaneous Layer Predicts the Grade of Leg Edema in Pregnant Women.

The technique most widely used to quantitatively measure leg edema is only a pitting edema method. It has recently become possible to digitize B-mode ultrasound images and accurately quantify their brightness using an image-analysis software program. The purpose of this study was to find new indices of the grade of leg skin, to study whether or not analyses of the subcutaneous layer of leg skin on ultrasound images using image-editing software program can be used to evaluate it and to digitize it. Images of 282 subcutaneous layers of leg skin in 141 pregnant women were obtained using a B-scan portable ultrasound device. Rectangular photographs (vertical: skin thickness; horizontal: width of probe) were obtained using an image-editing program, and the luminous intensity (pixel grayscale: 0-255) and thickness of the skin were calculated using a histogram. We investigated the correlation between these parameters and the grade of pitting edema (0-3). There was a significant positive correlation between the grade of pitting edema and the average luminous intensity value, its standard deviation, and the skin thickness (ρ = 0.36, ρ = 0.22, ρ = 0.51, p < 0.0001, respectively). In particular, there was strong positive correlation between the grade of pitting edema and both the total number of pixels in a rectangle × (multiplied by) the average luminous intensity value and the total number of pixels in a rectangle × the standard deviation of the average luminous intensity value (ρ = 0.58 and ρ = 0.59, p < 0.0001, respectively). We could quantitatively evaluate the grade of leg edema by analyzing ultrasound photographs of the subcutaneous layer of the leg skin using an image-editing software program and found new indices to digitize it.

Iridoids from Patrinia heterophylla and their anti-inflammatory activity.

A phytochemical investigation led to the isolation of five undescribed compounds (1-5) from the methanol extract of the rhizomes and roots of Patrinia heterophylla. The structures and configurations of these compounds were characterized by HRESIMS, ECD, and NMR data analyses. These compounds were assayed for their anti-inflammatory potential using LPS-stimulated BV-2 cells, of which compound 4 showed strong nitric oxide (NO) inhibitory effects with an IC50 of 6.48 µM. The potential anti-inflammatory mechanism was examined utilizing Western blotting and molecular docking. Further in vivo anti-inflammatory experiments revealed that compound 4 inhibited the NO production and reactive oxygen species in the zebrafish model.

Tetrabromobisphenol A and hexabromocyclododecane, brominated flame retardants, trigger endoplasmic reticulum stress and activate necroptosis signaling in PC12 cells.

Tetrabromobisphenol A (TBBPA) and hexabromocyclododecane (HBCD) are brominated flame retardants commonly used in a variety of industrial and consumer products. In this study, we performed RNA sequencing analysis of PC12 cells to clarify the mechanisms by which TBBPA and HBCD induce neurotoxicity. Differential expression analysis demonstrated that 636 and 271 genes were differentially expressed after TBBPA and HBCD treatment, respectively. Gene Ontology (GO) enrichment analysis revealed that genes annotated with the GO term "endoplasmic reticulum unfolded protein response" were upregulated in both TBBPA- and HBCD-treated groups. Furthermore, protein expression of endoplasmic reticulum stress markers, such as HSPA5 and DDIT3, as well as cleaved caspase-3, an apoptosis marker, were induced by TBBPA and HBCD. We also found that the cytotoxicity induced by TBBPA and HBCD was blocked by necrostatin-1, a necroptosis inhibitor, indicating the contribution of necroptosis. Our findings provide new insight into the mechanisms of toxicity induced by these chemicals.

Functional Analysis of Foxp3 and Its Mutants by Retroviral Transduction of Murine Primary CD4+ T Cells.

The transcription factor Foxp3/FOXP3 orchestrates regulatory T (Treg) cell development and function by interacting with numerous target genes and partner proteins. Functional analysis of naturally occurring or engineered Foxp3/FOXP3 mutations has provided important insights into how the complex Foxp3/FOXP3-centered molecular network operates. Here, we describe detailed protocols for retroviral transduction of murine primary conventional CD4+ T cells to determine the impacts of Foxp3 mutations on the Treg-cell-like phenotype and function conferred by Foxp3.

Whole-embryonic identification of maternal microchimeric cell types in mouse using single-cell RNA sequencing.

Even though the mother and the fetus of placental mammals are immunologically non-self with respect to one other, mutual exchange of small numbers of cells between them is known to occur. Maternal cells entering the fetus, called maternal microchimeric cells (MMc cells), are thought to be involved in different physiological phenomena, such as establishing immune tolerance, tissue repair, and the pathogenesis or deterioration of some inflammatory diseases and congenital malformations. While specific MMc cell types have been reported as associated with these phenomena, the contribution of MMc cells to these different outcomes remains unknown. As one possibility, we hypothesized that different embryos have differing repertoires of MMc cell types, leading to or biasing embryos toward different fates. To date, no studies have succeeded in identifying the MMc cell type repertoire of a single embryo. Accordingly, here, we isolated MMc cells from whole mouse embryos, determined their types, and analyzed their MMc cell type variability. By combining our previously established, whole-embryonic MMc isolation method with single-cell RNA sequencing, we successfully estimated the cell type repertoires of MMc cells isolated from 26 mouse embryos. The majority of MMc cells were immune-related cells, such as myeloid cells and granulocytes. We also detected stem cell-like MMc cells expressing proliferation marker genes and terminally differentiated cells. As hypothesized, we noted statistically significant inter-individual variation in the proportion of immune-related cells in the different embryos. We here successfully estimated MMc cell types in individual whole mouse embryos. The proportion of immune-related cells significantly differed among the individual embryos, suggesting that the variations are one of the potential mechanisms underlying the differing MMc-related physiological phenomena in offspring. These findings provide insight into cell-level epigenetics by maternal cells.

Measurement of Skin Thickness Using Ultrasonography to Test the Usefulness of Elastic Compression Stockings for Leg Edema in Pregnant Women.

Background: One of the most common treatments for leg edema during pregnancy is the use of compression stockings. The purpose of this study was to evaluate the objective effectiveness in pregnant women, by measuring the changes of skin thickness using ultrasonography. Methods: Pregnant women were diagnosed with leg edema using the pitting edema method at 36 weeks of gestation. Twenty-four pregnant women (48 legs) with leg edema spent time without wearing elastic stockings at 36−37 weeks of gestation. Then, they wore elastic stockings for one week at 37−38 weeks of gestation. We measured the grade of edema (from 0 to 3) and the skin thickness of the lower leg by portable ultrasonography at 36, 37, and 38 weeks of gestation (a before-and-after study). Results: In 24 pregnant women, thigh edema was not detected in any of the 48 legs before or after the use of elastic stockings. All 48 legs in 24 pregnant women had physiological lower leg edema, but not thigh edema. The average grade of pitting edema in each lower leg significantly decreased after using the stockings (36 weeks, 1.77 ± 0.85; 37 weeks, 1.79 ± 0.77; 38 weeks, 1.04 ± 0.74, p < 0.0001). In addition, the skin thickness of the lower legs was significantly decreased after the use of elastic stockings (36 weeks, 7.47 ± 2.45 mm; 37 weeks, 7.93 ± 2.83 mm; 38 weeks, 7.15 ± 2.35 mm, p < 0.0001). Conclusions: The wearing of elastic compression stockings on the lower legs is objectively effective for improving leg edema in pregnant women.

Structure, anti-tumor activity, and potential anti-tumor mechanism of a fungus polysaccharide from Fomes officinalis.

In our ongoing process of discovering bioactive macromolecules, a homogeneous polysaccharide (FOP80-1) was first purified from Fomes officinalis. FOP80-1 with molecular weight of 4560 Da was mainly composed of →3)-d-Galp-(1→, →4)-ß-d-Manp-(1→, →6)-α-d-Glcp-(1→, →3,6)-d-Glcp-(1→, and t--d-Glcp. Besides the structure features, the anti-tumor activity and potential mechanism of FOP80-1 were also investigated. The cellular and zebrafish experiments revealed that FOP80-1 inhibited tumor proliferation, invasion, and metastasis by increasing ROS, arresting cell cycle, inducing apoptosis, and suppressing angiogenesis. Corresponding to the inhibition of angiogenesis, the surface plasmon resonance (SPR) experiments revealed that FOP80-1 had good affinity with VEGF, a crucial protein to regulate angiogenesis. Molecular docking indicated that FOP80-1 could interact with the protein VEGF.

Anthocyanin-rich blackcurrant extract improves long-term memory impairment and emotional abnormality in senescence-accelerated mice.

Alzheimer's disease (AD) is associated with a progressive worsening in cognitive function, which is often accompanied by emotional disturbance. Recent studies revealed that anthocyanin-rich blackcurrant extract (BCE) can impart health benefits, but it is not known whether BCE is useful in the prevention and/or treatment of AD. Here, we examined the effects of BCE using a senescence-accelerated mouse prone 8 (SAMP8) model. Dietary BCE supplementation for 9 weeks was found to both improve the diminished long-term recognition memory and normalize the anxiety levels of SAMP8 mice. RNA sequencing demonstrated that dietary supplementation with anthocyanin-rich BCE significantly altered the gene expression profile in the hippocampus. According to enrichment analysis, genes regulated by BCE were related to cellular component terms such as "smooth endoplasmic reticulum," "axon," and "glutamatergic synapse." Real-time PCR verified alterations in the expression of AD-related genes. These findings indicate that anthocyanin-rich BCE may be valuable for the prevention and/or treatment of AD. PRACTICAL APPLICATIONS: Blackcurrant contains an abundance of polyphenols, especially anthocyanins. This study demonstrated that anthocyanin-rich BCE improves the long-term recognition memory impairment and emotional abnormality of SAMP8 mice, a mouse model characterized by several pathological features of AD. These findings indicate that anthocyanin-rich BCE may be a useful food supplement or ingredient for the prevention of AD.

A Narrative Review of the Effects of Citrus Peels and Extracts on Human Brain Health and Metabolism.

As life expectancy increases, age-associated diseases such as Alzheimer's disease (AD) become a major health problem. The onset of AD involves neurological dysfunction due to amyloid-ß accumulation, tau hyperphosphorylation, oxidative stress, and neuroinflammation in the brain. In addition, lifestyle-related diseases-such as dyslipidemia, diabetes, obesity, and vascular dysfunction-increase the risk of developing dementia. The world population ages, prompting the development of new strategies to maintain brain health and prevent the onset of dementia in older and preclinical patients. Citrus fruits are abundant polymethoxylated flavone and flavanone sources. Preclinical studies reported that these compounds have neuroprotective effects in models of dementia such as AD. Interestingly, clinical and epidemiological studies appear to support preclinical evidence and show improved cognitive function and reduced associated disease risk in healthy individuals and/or patients. This review summarizes the recent evidence of the beneficial effects of citrus peels and extracts on human cognition and related functions.

Periodontal acidification contributes to tooth pain hypersensitivity during orthodontic tooth movement.

Tooth movements associated with orthodontic treatment often cause tooth pain. However, the detailed mechanism remains unclear. Here, we examined the involvement of periodontal acidification caused by tooth movement in mechanical tooth pain hypersensitivity. Elastics were inserted between the first and second molars to move the teeth in Sprague-Dawley rats. Mechanical head-withdrawal reflex threshold to first molar stimulation and the pH of the gingival sulcus around the tooth were measured. The expression of acid-sensing ion channel 3 (ASIC3) in trigeminal ganglion neurons and phosphorylation of ASIC3 in the periodontal tissue were analyzed. The mechanical head-withdrawal reflex threshold to first molar stimulation and pH in the gingival sulcus decreased on day 1 after the elastic insertion. These decreases recovered to the sham level by buffering periodontal acidification. Periodontal inhibition of ASIC3 channel activity reversed the decreased mechanical head-withdrawal reflex threshold to first molar stimulation. On day 1 after elastic insertion, the tooth movement did not change the number of ASIC3 immunoreactive trigeminal ganglion neurons innervating the periodontal tissue but increased phosphorylated-ASIC3 levels in the periodontal tissue. Periodontal acidification induced by tooth movement causes phosphorylation of ASIC3, resulting in mechanical pain hypersensitivity in mechanically forced tooth.

Whole embryonic detection of maternal microchimeric cells highlights significant differences in their numbers among individuals.

During pregnancy in placental mammals, small numbers of maternal cells (maternal microchimeric cells, or MMc cells) migrate into the fetus and persist decades, or perhaps for the rest of their lives, and higher frequencies of MMc cells are reported to correlate with variety of phenomena, such as immune tolerance, tissue repair, and autoimmune diseases. While detection of these MMc cells is considered in all pregnancies, their frequency differs largely according to tissue type and disease cases, and it remains unclear whether the number of MMc cells differs significantly among embryos in normal pregnancies. Here, for the first time, we developed a whole embryonic detection method for MMc cells using transgenic mice and counted live MMc cells in each individual embryo. Using this technique, we found that the number of MMc cells was comparable in most of the analyzed embryos; however, around 500 times higher number of MMc cells was detected in one embryo at the latest stage. This result suggests that the number of MMc cells could largely differ in rare cases with unknown underlying mechanisms. Our methodology provides a basis for testing differences in the numbers of MMc cells among individual embryos and for analyzing differences in MMc cell type repertoires in future studies. These data could provide a hint toward understanding the mechanisms underlying the variety of apparently inconsistent MMc-related phenomena.

Evaluation of torque moment in esthetic brackets from bendable alloy wires.

OBJECTIVES: To examine the torque moment that occurs between esthetic brackets and bendable alloy (stainless steel [SS], titanium-molybdenum [Ti-Mo], and titanium-niobium [Ti-Nb]) wires. MATERIALS AND METHODS: This study examined ceramic (CR), zirconium oxide (ZC), polycarbonate (PC), and conventional metallic brackets (MT) (upper, 0.018-inch and 0.022-inch slots) combined with SS, Ti-Mo, and Ti-Nb wires using elastic module ligation. The torque moments delivered by various wire and bracket combinations were measured using a torque gauge apparatus. The wire torque angles at 5-40° were examined. RESULTS: The torque value increased in the order of CR, ZC, MT, and PC brackets for both 0.018-inch and 0.022-inch slots. The fracture points of the CR and ZC brackets combined with SS and Ti-Mo wires were approximately more than 30° and 35°, respectively. No fracture points were detected in the combination of ZC brackets and Ti-Nb wires. CONCLUSIONS: The current study identified the material characteristics of CR, ZR, and PC brackets during torque tooth movements. The present results demonstrate a characteristic combined effect between different esthetic brackets and bendable alloy wires.

Preparation of cerium molybdates and their antiviral activity against bacteriophage Φ6 and SARS-CoV-2.

Two cerium molybdates (Ce2Mo3O12 and γ-Ce2Mo3O13) were prepared using either polymerizable complex method or hydrothermal process. The obtained powders were almost single-phase with different cerium valence. Both samples were found to have antiviral activity against bacteriophage Φ6. Especially, γ-Ce2Mo3O13 exhibited high antiviral activity against both bacteriophage Φ6 and SARS-CoV-2 coronavirus, which causes COVID-19. A synergetic effect of Ce and molybdate ion was inferred along with the specific surface area as key factors for antiviral activity.

Effects of cerium and tungsten substitution on antiviral and antibacterial properties of lanthanum molybdate.

Powders of cerium (Ce)-substituted and tungsten (W)-substituted La2Mo2O9 (LMO) were prepared using polymerizable complex method. Their antiviral and antibacterial performances were then evaluated using bacteriophage Qß, bacteriophage Φ6, Escherichia coli, and Staphylococcus aureus. The obtained powders, which were almost single-phase, exhibited both antiviral and antibacterial properties. Effects of dissolved ions on their antiviral activity against bacteriophage Qß were remarkable. A certain contribution of direct contact to the powder surface was also inferred along with the dissolved ion effect for antiviral activity against bacteriophage Φ6. Dissolved ion effects and pH values suggest that both Mo and W are in the form of polyacids. Antiviral activity against bacteriophage Φ6 was improved by substituting Ce for La in LMO. Similarly to LMO, Ce-substituted LMO exhibited hydrophobicity. Inactivation of alkaline phosphatase enzyme proteins was inferred as one mechanism of the antiviral and antibacterial activities of the obtained powders.

An evaluation of the genotoxicity and subchronic toxicity of the peel extract of Ponkan cultivar 'Ohta ponkan' (Citrus reticulata Blanco) that is rich in nobiletin and tangeretin with anti-dementia activity.

Nobiletin and tangeretin are major components of polymethoxylated flavones in the peels of citrus fruits such as Citrus reticulata. Because nobiletin and tangeretin have attracted attention due to their beneficial health properties, citrus peel extracts, in which they are concentrated, have the potential to serve as a functional food ingredient to prevent diseases. In this study, a series of toxicological studies on the peel extract of Ponkan cultivar 'Ohta ponkan' (Citrus reticulata Blanco), was conducted. No mutagenic activity was observed in a bacterial reverse mutation test, whereas chromosomal aberrations were induced in an in vitro mammalian chromosomal aberration test. No genotoxicity was observed in an in vivo mammalian micronucleus test. In a 90-day study at daily doses of 54, 180, or 540 mg/kg body weight (bw)/day, hyaline droplet nephropathy, which specifically occurs in adult male rats, was observed in males of 540 mg/kg bw/day group. No other adverse effects were observed in the 90-day study. The no adverse effect level in the 90-day study was considered to be 540 mg/kg bw/day for female rats and less than 540 mg/kg bw/day for male rats.

Identification of Smad-dependent and -independent signaling with transforming growth factor-ß type 1/2 receptor inhibition in palatogenesis.

TGF-ß signaling is one of important function during palatal fusion. Three types of TGF-ß receptor (TßR1, TßR2, and TßR3) have been identified, and play essential roles in mechanisms leading to palatal fusion. However, the balance between Smad-dependent/-independent signaling during palatal fusion with inhibited TßR1/2 functions is not fully understood. The objective of this study was to investigate palatal fusion via TGF-ß signaling when TßR1 and TßR2, but not TßR3, were inhibited. In addition, the present study examined the functional balance between Smad-dependent/-independent signaling and related gene expression. Palatal organ cultures were treated with TßR1/2 inhibitor in vitro. Control palates were cultured without inhibitor. We observed histological phenotype of palatal fusion, and evaluation of expression pattern by Western blot or real time RT-PCR. Palatal organ cultures treated with the inhibitor did not fuse and the medial edge epithelium remained at embryonic 13 day +72 h in culture. The inhibitor decreased TßR1 and TßR2 expression by approximately 90%, but did not affect TßR3 expression. The expression of p-Smad2 and p-Smad3 was significantly decreased in treated palates compared with controls. The expression of p-Smad4 was slightly decreased in treated palates compared with controls. Smad-independent signaling was also affected by the inhibitor; p-ERK, p-JNK, and p-p38 expressions was significantly reduced in treated palates compared with controls. The expression of transcription factors (Runx1 and Msx1) and extracellular matrix proteins (MMP2/13) was also significantly decreased by inhibitor exposure. Treatment with TßR1/2 inhibitor altered the patterns of the Smad-dependent and -independent signaling pathways during palatal fusion.