The hyperphosphorylation of α-synuclein (α-Syn) at serine 129 is associated with in its aggregation, culminating in the formation of Lewy bodies (LBs) in Parkinson's disease (PD) and dementia with LBs. Plasma homocysteine (Hcy) levels are typically high in PD, particularly in those treated with Levodopa. Therefore, we aimed to investigate the effects of Hcy on phosphorylation and aggregation of α-Syn. The human neuroblastoma cell line 3D5 expressing wild-type α-Syn under the control of a tetracycline-off system was treated with Hcy, and aggregation and phosphorylation of α-Syn were examined. Hcy was cytotoxic to 3D5 cells, and Hcy induced cell shrinkage at a concentration >100 ‍μM. Treatment with Hcy upregulated the levels of α-Syn, increased its phosphorylation at serine 129, and increased casein kinase 2A. Moreover, Hcy treatment significantly increased the level of aggregated form of α-Syn, including oligomers in 3D5 cells. However, folate significantly reversed the Hcy-mediated increase in the levels of total α-Syn and its phosphorylation. In conclusion, Hcy enhances the total α-Syn level, and phosphorylated and oligomeric α-Syn formation, thereby promoting LBs formation.

AIM:The aim was to explore the preventive effects of smartphone cognitive behavioral therapy (CBT) in addressing depression and anxiety in patients with recurrent pregnancy loss (RPL) due to lack of such previous studies. METHODS:This was an exploratory decentralized, parallel-group, multicenter, open, individually randomized, fully factorial trial. A total of 432 patients with RPL were randomized and patients who completed the psychoeducation component of the app within 2 weeks of consent were included. Five CBT components-namely, self-monitoring, behavioral activation, cognitive restructuring, assertion training, and problem solving-were randomly assigned as present or absent, resulting in 25 = 32 combinations. The primary outcomes were changes in Patient Health Questionnaire-9 and Generalized Anxiety Disorder-7 scores at weeks 8 and 12 from baseline. RESULTS:A total of 419 patients were analyzed. The presence or absence of specific CBT components did not significantly affect patient outcomes. However, patients experienced a statistically significant overall reduction in depressive symptoms from baseline to weeks 8 and 12. Pre-post effect sizes for overall improvement ranged from -0.30 to -0.38 at week 8 and from -0.22 to -0.33 at week 12. No significant overall reduction in anxiety symptoms was observed. CONCLUSION:The CBT application significantly reduced depressive symptoms in patients with RPL; however, no specific CBT component showed superior effects. The findings did not indicate efficacy in comparison to the pure treatment-as-usual group. Further research is required to enhance its impact on anxiety and refine the selection and delivery of individual CBT components.

Homeobox transcription factors CDX1 and CDX2 (hereafter, CDX1/2) play key roles in determining the identity of intestinal epithelial cells and regulating their stem cell functions. However, the role of CDX1/2 in regulating colon cancer stemness and the underlying mechanisms are unclear. Here, we show that complete loss of Cdx1 or concurrent loss of Cdx1/2 increased the stemness and malignancy of intestinal tumors. Consistently, CDX1/2 reduced the expression of cancer stemness-related genes, including LGR5. CDX1/2 bound to the downstream region of the LGR5 transcription start site (TSS), a region where β-catenin also binds. Despite increased H3 acetylation and an open chromatin structure, CDX1/2 reduced the occupancy of DRB sensitivity-inducing factor (DSIF), RNA polymerase II-associated factor 1 (PAF1), and RNA polymerase II (Pol II) complexes around the LGR5 TSS. Through their homeodomains, CDX1/2 inhibited the β-catenin-facilitated formation of active Pol II complexes containing DSIF and PAF1 complexes by preventing the interaction between β-catenin and these complexes, in an additive manner. Our findings suggest that CDX1/2 cooperatively suppressed colonic tumorigenesis and cancer stemness by antagonizing β-catenin via the DSIF and PAF1 complexes. Additionally, DSIF and PAF1 complexes acted as transcriptional platforms that integrated and funneled both tumor-suppressive and oncogenic signals into the expression of genes that control colon cancer stemness.

An investigation into the acute toxicity of mechanically fibrillated cellulose nanofibers (fib-CNFs), with a fiber length ranging from 500 to 600 nm, was conducted in accordance with the OECD TG412 guidelines. In this study, rats were exposed to fib-CNFs via nasal inhalation for 6 h daily over a 28-day period. The highest exposure concentration was set at 35 mg/m3, with intermediate and low concentrations at 7.0 mg/m3 and 1.5 mg/m3, respectively. No significant differences were observed in body weight, hematological parameters, or biochemical profiles between the fib-CNF-exposed groups and the control group. However, the histopathological examination of lung tissue revealed elevated macrophage counts in both the alveolar spaces and lymph nodes, accompanied by a significant increase in lung weight. The most severe effects were observed in the high-concentration group, while the low-concentration group exhibited only mild inflammatory changes. Based on these findings, the no observable adverse effect level (NOAEL) for the acute toxicity of fib-CNFs is estimated to be below 1.5 mg/m3.

The impact of oral administration of mechanically fibrillated cellulose nanofibers (fib-CNF), a commonly used nanofiber, on toxicity and health remains unclear, despite reports of the safety and beneficial effects of chitin-based nanofibers. Thus, evaluating the oral toxicity of fib-CNF in accordance with OECD Test Guideline 407 (TG407) is essential. This study aimed to assess the safety of orally administered fib-CNF through an acute toxicity study in rats, following the OECD TG407 guidelines for 4 weeks. CNF “BiNFi-s” FMa-10005, derived from mechanically fibrillated pulp cellulose, was administered via gavage to male and female Crl:CD(SD) rats at doses of 50, 150, 500, and 1000 mg/kg/day for 28 days, with a control group receiving water for injection. The study evaluated the toxic effects of repeated administration, and the rats were monitored for an additional 14 days post-administration to assess recovery from any toxic effects. The results showed no mortality in either sex during the administration period, and no toxicological effects related to the test substance were observed in various assessments, including general condition and behavioral function observations, urinalysis, hematological examination, blood biochemical examination, necropsy findings, organ weights, and histopathological examination. Notably, only female rats treated with 1000 mg/kg/day of CNF exhibited a consistent reduction in body weight during the 14-day recovery period after the end of treatment. They also showed a slight decrease in pituitary and liver weights. However, hematological and blood biochemical tests did not reveal significant differences, suggesting a potential weight-suppressive effect of CNF ingestion.

Spleen tyrosine kinase (Syk), a non-receptor-type tyrosine kinase, has a wide range of physiological functions. A possible role of Syk in Alzheimer's disease (AD) has been proposed. We evaluated the localization of Syk in the brains of patients with AD and control participants. Human neuroblastoma M1C cells harboring wild-type tau (4R0N) were used with the tetracycline off (TetOff) induction system. In this model of neuronal tauopathy, the effects of the Syk inhibitors—BAY 61–3606 and R406—on tau phosphorylation and oligomerization were explored using several phosphorylated tau-specific antibodies and an oligomeric tau antibody, and the effects of these Syk inhibitors on autophagy were examined using western blot analyses. Moreover, the effects of the Syk inhibitor R406 were evaluated in vivo using wild-type mice. In AD brains, Syk and phosphorylated tau colocalized in the cytosol. In M1C cells, Syk protein (72 kDa) was detected using western blot analysis. Syk inhibitors decreased the expression levels of several tau phosphoepitopes including PHF-1, CP13, AT180, and AT270. Syk inhibitors also decreased the levels of caspase-cleaved tau (TauC3), a pathological tau form. Syk inhibitors increased inactivated glycogen synthase kinase 3β expression and decreased active p38 mitogen-activated protein kinase expression and demethylated protein phosphatase 2 A levels, indicating that Syk inhibitors inactivate tau kinases and activate tau phosphatases. Syk inhibitors also activated autophagy, as indicated by increased LC3II and decreased p62 levels. In vivo, the Syk inhibitor R406 decreased phosphorylated tau levels in wild-type mice. These findings suggest that Syk inhibitors offer novel therapeutic strategies for tauopathies, including AD.

Mutations in APC, found in 80% of colon caner, enhance β-catenin stabilization, which is the initial step of colonic tumorigenesis. However, the core transcriptional mechanism underlying the induction of colon cancer stemness by stable β-catenin remains unclear. Here, we found that inducible inhibition of β-catenin suppressed elongation of Pol II and RNA polymerase-associated factor 1 complex (PAF1C) around the transcription start site (TSS) of LGR5. Moreover, stable β-catenin enhanced the formation of active Pol II complex cooperatively with CDC73 and CDK9 by facilitating the recruitment of DRB sensitivity-inducing factor (DSIF) and negative elongation factor (NELF) complexes to the Pol II complex. Subsequently, stable β-catenin facilitated the formation of the Pol II–DSIF–PAF1C complex, suggesting that stable β-catenin induces cancer stemness by stimulating active Pol II complex through NELF and PAF1C. Furthermore, NELF or PAF1C inhibition recapitulated the changes in cancer stemness-related gene expression induced by the inhibition of stable β-catenin and suppressed colon cancer stemness. Additionally, the chemical inhibition of CDK12 (a downstream transcription CDK of PAF1C) suppressed colon cancer stemness. These results suggest that NELF and PAF1C are the core transcriptional machineries that control expression of colon cancer stemness-inducing genes and may be therapeutic targets for colon cancer.

Mechanically fibrillated cellulose nanofibers, known as fib-CNF (fiber length: 500 nm; diameter: 45 nm), are used in composites and as a natural thickener in foods. To evaluate their safety, we conducted a 28-day study in mice with inhalation exposure at 0.2 mg/body and oral administration of 400 mg/kg/day. Inhalation exposure to fib-CNF caused transient weight loss, changes in blood cell counts, and increased lung weights. These changes were attributed to adaptive responses. The oral administration of fib-CNF for 28 days resulted in no apparent toxic effects except for a slight decrease in platelet counts. The fib-CNF administration using the protocols studied appears to be safe in mice.

Supplementary Methods, Figure Legends 1-12 from Suppression of Colonic Polyposis by Homeoprotein CDX2 through its Nontranscriptional Function that Stabilizes p27Kip1

Supplementary Figure 4 from Suppression of Colonic Polyposis by Homeoprotein CDX2 through its Nontranscriptional Function that Stabilizes p27Kip1

Supplementary Methods, Figure Legends 1-12 from Suppression of Colonic Polyposis by Homeoprotein CDX2 through its Nontranscriptional Function that Stabilizes p27^Kip1

No genetic association with recurrent pregnancy loss (RPL) caused by embryonic aneuploidy has been found. Recent studies have indicated that the common genetic variant rs2305957, surrounding the PLK4 gene, contributes to mitotic-origin aneuploidy risk during human early embryo development. The decrease in meiosis-specific cohesin causes predivision of sister chromatids in the centromere and chromosome segregation errors. STAG3 is a component of cohesin and is a meiosis-specific gene. Our case-control study included 184 patients with RPL whose previous products of conception (POC) exhibited aneuploidy and 190 fertile control women without a history of miscarriage. We performed a genetic association study to examine the genotype distribution at PLK4 (rs2305957) and STAG3 in patients with RPL caused by aneuploidy compared with controls. Regarding STAG3, SNPs with a minor allele frequency (MAF) threshold > 0.05 that were predicted to be binding sites of transcription factors and that showed significant associations in expression quantitative trait locus (e-QTL) analysis were selected. No significant differences in the MAF or distribution in any model of PLK4 (rs2305957) and 5 selected tag SNPs in STAG3 were found between the patients and controls. A further genome-wide association study is needed since a combination of genetic risk alleles might be useful in predicting future age-dependent RPL caused by aneuploidy.

Autophagy, a system for the bulk degradation of intracellular components, is essential for homeostasis and the healthy physiology and development of cells and tissues. Its deregulation is associated with human disease. Thus, methods to modulate autophagic activity are critical for analysis of its role in mammalian cells and tissues. Here we report a method to inhibit autophagy using a mutant variant of the protein ATG7, a ubiquitin E1-like enzyme essential for autophagosome formation. During autophagy, ATG7 activates the conjugation of LC3 (ATG8) with phosphatidylethanolamine (PE) and ATG12 with ATG5. Human ATG7 interactions with LC3 or ATG12 require a thioester bond involving the ATG7 cysteine residue at position 572. We generated TetOff cells expressing mutant ATG7 protein carrying a serine substitution of this critical cysteine residue (ATG7C572S). Because ATG7C572S forms stable intermediate complexes with LC3 or ATG12, its expression resulted in a strong blockage of the ATG-conjugation system and suppression of autophagosome formation. Consequently, ATG7C572S mutant protein can be used as an inhibitor of autophagy.

We have analyzed Hall effect in a thin-film transistor by actual experiments and device simulation. Particularly in this presentation, we show the sensitivity dependence on the applied voltage. These behaviors can be explained by the electric field in the channel layer for the operation regions. Additionally, the carrier mobilities and densities calculated using transistor equation and Hall effect are compared.

Magnetoresistive (MR) sensors made with amorphous In-Ga-Zn-O (a-IGZO) that have significant conventional or inverse MR effects at room temperature were fabricated. A high-temperature a-IGZO MR thin-film transistor exhibited a giant inverse MR effect of up to -69% at 25 mT. The detailed measurement results are reported and themechanism for generation of theMR effect is discussed.

We have analyzed the Hall effect in a thin-film transistor. First, we investigated the dependence of the Hall voltage on the gate voltage. In the subthreshold region, a relatively large Hall voltage is generated regardless of the magnetic field. This is due to the zigzag path of the current flow. In the saturation region, as the gate voltage increases, the Hall voltage also increases, which is dependent on the magnetic field, and in the linear region, it is saturated. Next, we investigated the dependence on the drain voltage. In the linear region, as the drain voltage increases, the Hall voltage also increases, and in the saturation region, it is saturated again. These behaviors can be explained by the electric field in the channel layer calculated using device simulation.