Regulation of Hippo-YAP signaling axis by Isoalantolactone suppresses tumor progression in cholangiocarcinoma.

Cholangiocarcinoma (CCA) is a devastating malignancy characterized by aggressive tumor growth and limited treatment options. Dysregulation of the Hippo signaling pathway and its downstream effector, Yes-associated protein (YAP), has been implicated in CCA development and progression. In this study, we investigated the effects of Isoalantolactone (IALT) on CCA cells to elucidate its effect on YAP activity and its potential clinical significance. Our findings demonstrate that IALT exerts cytotoxic effects, induces apoptosis, and modulates YAP signaling in SNU478 cells. We further confirmed the involvement of the canonical Hippo pathway by generating LATS1/LATS2 knockout cells, highlighting the dependence of IALT-mediated apoptosis and YAP phosphorylation on the Hippo-LATS signaling axis. In addition, IALT suppressed cell growth and migration, partially dependent on YAP-TEAD activity. These results provide insights into the therapeutic potential of targeting YAP in CCA and provide a rationale for developing of YAP-targeted therapies for this challenging malignancy.

Peptide-Decorated Microneedles for the Detection of Microplastics.

The escalating utilization of plastics in daily life has resulted in pervasive environmental pollution and consequent health hazards. The challenge of detecting and capturing microplastics, which are imperceptible to the naked eye, is exacerbated by their diminutive size, hydrophobic surface properties, and capacity to absorb organic compounds. This study focuses on the application of peptides, constituted of specific amino acid sequences, and microneedles for the rapid and selective identification of microplastics. Peptides, due to their smaller size and greater environmental stability compared with antibodies, emerge as a potent solution to overcome the limitations inherent in existing detection methodologies. To immobilize peptides onto microneedles, this study employed microneedles embedded with gold nanorods, augmenting them with sulfhydryl (SH) groups at the peptides' termini. The sensor developed through this methodology exhibited efficient peptide binding to the microneedle tips, thereby facilitating the capture of microplastics. Raman spectroscopy was employed for the detection of microplastics, with the results demonstrating successful attachment to the microneedles. This novel approach not only facilitates localized analysis but also presents a viable strategy for the detection of microplastics across diverse environmental settings.

The LKB1-TSSK1B axis controls YAP phosphorylation to regulate the Hippo-YAP pathway.

The Hippo pathway's main effector, Yes-associated protein (YAP), plays a crucial role in tumorigenesis as a transcriptional coactivator. YAP's phosphorylation by core upstream components of the Hippo pathway, such as mammalian Ste20 kinase 1/2 (MST1/2), mitogen-activated protein kinase kinase kinase kinases (MAP4Ks), and their substrate, large tumor suppressor 1/2 (LATS1/2), influences YAP's subcellular localization, stability, and transcriptional activity. However, recent research suggests the existence of alternative pathways that phosphorylate YAP, independent of these core upstream Hippo pathway components, raising questions about additional means to inactivate YAP. In this study, we present evidence demonstrating that TSSK1B, a calcium/calmodulin-dependent protein kinase (CAMK) superfamily member, is a negative regulator of YAP, suppressing cellular proliferation and oncogenic transformation. Mechanistically, TSSK1B inhibits YAP through two distinct pathways. Firstly, the LKB1-TSSK1B axis directly phosphorylates YAP at Ser94, inhibiting the YAP-TEAD complex's formation and suppressing its target genes' expression. Secondly, the TSSK1B-LATS1/2 axis inhibits YAP via phosphorylation at Ser127. Our findings reveal the involvement of TSSK1B-mediated molecular mechanisms in the Hippo-YAP pathway, emphasizing the importance of multilevel regulation in critical cellular decision-making processes.

Effects of Compression of the Ulnar Artery on the Radial Artery Catheterization.

Background: The study was designed to evaluate the effects of compression of the ulnar artery on blood flow (BF) and internal cross-sectional area (CSAi) of the radial artery. We also evaluated the success rate and time of successful ultrasound-guided radial artery catheterization at the first attempt with or without compression of the ulnar artery. Methods: Patients were randomly allocated to the Compression group or Standard group to be treated with or without the application of ulnar artery compression, respectively. Hemodynamic stability was confirmed, and ultrasound-guided radial artery catheterization was performed. In the Compression group, an assistant compressed the ulnar artery at 5 cm above the wrist crease and the catheterization was performed after the loss of the distal ulnar artery BF. In the Standard group, the catheterization was performed without compression of the ulnar artery. Before and after the catheterization, BF and CSAi of the radial artery were evaluated. Success rate and time to successful catheterization at the first attempt were recorded. Results: BF and CSAi of the radial artery were similar in the two groups (37.5 [19.3−66] vs. 37.0 [20.6−53.7] mL/min, respectively, p = 0.63; 4.0 [4.0−6.0] vs. 4.0 [3.0−5.0] mm2, respectively, p = 0.095). In the Compression group, BF and CSAi were changed to 80.9 [35.9−128.5] mL/min (p < 0.001) and 5.0 [4.0−7.0] mm2 (p < 0.001), respectively, after compression of the ulnar artery. There was a trend that the success rate of ultrasound-guided radial artery catheterization at the first attempt was higher in the Compression group than in the Standard group (58/59 vs. 53/59, respectively, p = 0.05), although the difference was not statistically significant. However, the time to successful ultrasound-guided radial artery catheterization at the first attempt was significantly shorter in the Compression group than in the Standard group (34 [27−41] s vs. 46 [36−60] s, p < 0.001). Conclusion: Compression of the ulnar artery augmented BF and CSAi of the radial artery. It resulted in a significantly shorter success time for ultrasound-guided radial artery catheterization at the first attempt.

Phosphorylation analysis of the Hippo-YAP pathway using Phos-tag.

Phosphorylation is an essential regulatory mechanism in cells that modifies diverse substrates, such as proteins, carbohydrates, lipids, and nucleotides. Protein phosphorylation regulates function, subcellular localization, and protein-protein interactions. Protein kinases and phosphatases catalyze this reversible mechanism, subsequently influencing signal transduction. The dysregulation of protein phosphorylation leads to many diseases, such as cancer, neurodegenerative diseases, and metabolic diseases. Therefore, analyzing the phosphorylation status and identifying protein phosphorylation sites are critical for elucidating the biological functions of specific phosphorylation events. Unraveling the critical phosphorylation events associated with diseases and specific signaling pathways is promising for drug discovery. To date, highly accurate and sensitive approaches have been developed to detect the phosphorylation status of proteins. In this review, we discuss the application of Phos-tag to elucidate the biological functions of Hippo pathway components, with emphasis on the identification and quantitation of protein phosphorylation under physiological and pathological conditions. SIGNIFICANCE: We here provide a comprehensive overview of Phos-tag technique-based strategies to identify phosphorylated proteins at the cellular level in the Hippo-YAP pathway that comprises a major driving force for cellular homeostasis. We clarify the links of applying Phos-tag in elucidating the biological functions of the Hippo pathway components with particular attention to the identification and quantitation of protein phosphorylation under physiological and pathological conditions. We believe that our paper will make a significant contribution to the literature because these detailed phosphorylation modifications and functional diversity of the Hippo pathway components in physiological and pathological processes are only beginning to come to the fore, highlighting the potential for discovering new therapeutic targets. Moreover, this line of research can provide further insight into the inextricable link between phos-tag applications as a molecular tool and cellular signaling modality, offering new directions for an integrated research program toward understanding cellular regulation at the molecular level. Given the broad research and practical applications, we believe that this paper will be of interest to the readership of your journal.

Clinical impact of medication adherence on 10-year cardio-cerebrovascular mortality in newly diagnosed hypertensive patients.

The purpose of this study is to evaluate the impact of medication adherence on cardio-cerebrovascular (CCV) mortality in newly diagnosed hypertensive patients. The authors retrospectively reviewed data from 20,836 patients who newly diagnosed hypertension from January 1, 2003 to December 31, 2005. Medication adherence was calculated from the compliance ratio (CR) during the first year after the diagnosis of hypertension. CCV mortality for 10 years was assessed according to the presence or absence of complications of hypertension. The risk of CCV death was significantly reduced in the CR ≥ 70% group than in the CR < 70% group (hazard ratio, 0.70; p = .004) for 10 years. In the patients without complications, the risk of CCV death was significantly lower in the CR ≥ 70% group than in the CR < 70% group (hazard ratio, 0.56; p = .014). However, in patients with complications, there was no significant difference in risk of CCV death between the CR ≥ 70% group and the CR < 70% group (hazard ratio, 0.79; p = .100). Only the CR ≥ 90% group had a significantly lower risk of CCV death (hazard ratio, 0.56; p < .001) for those with complications. Medication adherence is significantly associated with CCV mortality during 10 years in newly diagnosed hypertensives patients. Patients with complications of hypertension have to continue a high adherence rate (CR ≥ 90) for better long-term clinical outcomes.

Extracts of Perilla frutescens var. Acuta (Odash.) Kudo Leaves Have Antitumor Effects on Breast Cancer Cells by Suppressing YAP Activity.

Yes-associated protein (YAP)/WW domain-containing transcription factor (TAZ) is critical for cell proliferation, survival, and self-renewal. It has been shown to play a crucial oncogenic role in many different types of tumors. In this study, we investigated the antitumor effect of the extracts of Perilla frutescens var. acuta (Odash.) Kudo leaves (PLE) on Hippo-YAP/TAZ signaling. PLE induced the phosphorylation of YAP/TAZ, thereby inhibiting their activity. In addition, the treatment suppresses YAP/TAZ transcriptional activity via the dissociation of the YAP/TAZ-TEAD complex. To elucidate the molecular mechanism of PLE in the regulation of YAP activity, we treated WT and cell lines with gene knockout (KO) for Hippo pathway components with PLE. The inhibitory effects of PLE on YAP-TEAD target genes were significantly attenuated in LATS1/2 KO cells. Moreover, we found the antitumor effect of PLE on MDA-MB-231 and BT549, both of which are triple-negative breast cancer (TNBC) cell lines. PLE reduced the viability of TNBC cells in a dose-dependent manner and induced cell apoptosis. Further, PLE inhibited the migration ability in MDA-MB-231 cells. This ability was weakened in YAP and TEAD-activated clones suggesting that the inhibition of migration by PLE is mainly achieved by regulating YAP activity. Taken together, the results of this study indicate that PLE suppressed cell growth and increased the apoptosis of breast cancer (BC) cells via inactivation of YAP activity in a LATS1/2-dependent manner.

Hesperetin inhibit EMT in TGF-ß treated podocyte by regulation of mTOR pathway.

Renal fibrosis is one of the characteristic features of chronic kidney disease (CKD). Fibrotic change not only impairs the filtration function of the kidney but is also recognized as a marker of end-stage renal disease (ESRD). The epithelial to mesenchymal transition (EMT) is known to play a role in embryonic development and organ formation, but it is getting much attention for its pathological role in the invasion and metastasis of carcinoma. Recently, it has also been reported that EMT plays a role in the formation of fibrosis during chronic inflammation. EMT contribute to the development of the fibrosis in CKD. Moreover, glomerular podocytes and tubular epithelial cells can also undergo mesenchymal transition in CKD. Hesperetin is a flavonoid present in citrus and is well known for its antioxidant and anti-inflammatory properties. In this study, we investigated the effects of hesperetin on the EMT-elicited podocytes. First, we generated an EMT model by treating transforming growth factor (TGF)-ß1, a potent inducer of EMT to the podocytes. TGF-ß1 decreased the expression of epithelial markers such as nephrin, zonula occludens-1 (ZO-1), while it increased the mesenchymal markers, including fibronectin (FN), vimentin, and α-smooth muscle actin (α-SMA) in the podocytes. Hesperetin suppressed EMT-like changes elicited by TGF-ß1. Interestingly, hesperetin did not interfere with the Smad signaling-the classical TGF-ß signaling-pathway, which was confirmed by the experiment with smad 2/3 -/- podocytes. Instead, hesperetin suppressed EMT-like changes by inhibiting the mTOR pathway-one of the alternative TGF-ß signaling pathways. In conclusion, hesperetin has a protective effect on the TGF-ß1 elicited EMT-like changes of podocytes through regulation of mTOR pathway. It could be a good candidate for the suppression of kidney fibrosis in various CKD.

Role of the Hippo Pathway in Fibrosis and Cancer.

The Hippo pathway is the key player in various signaling processes, including organ development and maintenance of tissue homeostasis. This pathway comprises a core kinases module and transcriptional activation module, representing a highly conserved mechanism from Drosophila to vertebrates. The central MST1/2-LATS1/2 kinase cascade in this pathway negatively regulates YAP/TAZ transcription co-activators in a phosphorylation-dependent manner. Nuclear YAP/TAZ bind to transcription factors to stimulate gene expression, contributing to the regenerative potential and regulation of cell growth and death. Recent studies have also highlighted the potential role of Hippo pathway dysfunctions in the pathology of several diseases. Here, we review the functional characteristics of the Hippo pathway in organ fibrosis and tumorigenesis, and discuss its potential as new therapeutic targets.

The moderating effect of heart rate variability on the relationship between alpha asymmetry and depressive symptoms.

Electroencephalographic (EEG) research has suggested relatively reduced brain activity in the left frontal and right posterior region trait-markers of depression. However, inconsistent results have been reported. Based on previous studies reporting the heart rate variability (HRV) as an index of emotional regulation, this study makes a novel investigation of the role of heart rate variability (HRV) as a moderator in the relationship between frontal and parietal alpha asymmetry and depression. Resting EEG (eyes open) was recorded in 38 patients with MDD and 34 healthy subjects. Frontal and parietal alpha asymmetries were calculated at total (8-12 Hz), high (10-12 Hz), and low (8-10 Hz) alpha frequency bands. Three vagally mediated HRV (vmHRV) components (LF, HF, and the LF/HF ratio) were calculated in the frequency domain. Relatively greater right parietal alpha activity significantly predicted the severity of depression only when HF was low (or the LF/HF ratio was high) at low alpha frequency band. The interaction effect of parietal alpha asymmetry and vmHRV remained significant after including anxiety score as a covariate. No moderation effect of vmHRV was found for frontal sites and other frequency bands, as well as healthy subjects. These findings suggest that vmHRV moderates the association between parietal alpha asymmetry at low frequency band and depression for MDD patients. We suggest that the interaction between parietal alpha asymmetry and vmHRV may be a biomarker of MDD.