Alcohol stimulates the proliferation of mouse small intestinal epithelial cells via Wnt signaling.

The intestinal epithelium is one of the fastest renewing tissues in mammals and is a barrier against toxic substances such as alcohol. Excessive alcohol can induce intestinal damage leading to intestinal bowel diseases. Thus, the control of small intestinal epithelial cell (IEC) regeneration is thought to be important for homeostasis in response to epithelium damage. However, reports on how epithelial cells respond to small intestinal damage are scarce. We investigated the effects of alcohol consumption on small intestinal epithelial cells of mice. To verify that alcohol altered the small intestinal epithelium, we used 8-10 weeks old male C57BL/6J mice for models of chronic and binge alcohol consumption (the NIAAA model) in addition to an organoid model. Alcohol promoted the proliferative activity of IECs and intestinal stem cells (ISCs) in intestinal crypts. Alcohol consumption increased expression of the proliferation marker cyclin D1 and activated the p44/42 MAPK (Erk1/2) signaling pathway in small intestinal epithelial cells. The Wnt target genes were markedly increased in IECs from alcohol-treated mice. In the small intestinal organoid model exposed to alcohol, the organoid area and numbers of buds increased with alcohol concentrations up to 0.5% similar to in vivo observations. These results suggest that alcohol consumption stimulates the proliferation of small intestinal epithelial cells via Wnt signaling.

Widdrol-induced lipolysis is mediated by PKC and MEK/ERK in 3T3-L1 adipocytes.

Obesity is a serious medical condition causing various diseases such as heart disease, type-2 diabetes, and cancer. Fat cells (adipocytes) play an important role in the generation of energy through hydrolysis of lipids they accumulate. Therefore, induction of lipolysis (breakdown of lipids into fatty acids and glycerol), is one of the ways to treat obesity. In the present study, we investigated the lipolytic effect of widdrol in 3T3-L1 adipocytes and its mechanism. Widdrol considerably increased the amount of glycerol released from 3T3-L1 adipocytes into the medium in a time- and dose-dependent manner. To determine the mechanism of this effect, we investigated the alterations in glycerol release and protein expression in 3T3-L1 adipocytes treated with widdrol alone or widdrol and inhibitors of proteins involved in the cAMP-dependent pathway or cAMP-independent PKC-MAPK pathway, which are known to induce lipolysis in adipocytes. The adenylyl cyclase inhibitor SQ-22536, PLA2 inhibitor dexamethasone, PI3K inhibitor wortmannin, and PKA inhibitor H-89, which were used to investigate the involvement of the cAMP-dependent pathway, did not affect the lipolytic effect of widdrol. Widdrol-induced phosphorylation of PKC, MEK, and ERK, which are related to the PKC-MAPK pathway, and their phosphorylation was inhibited by their inhibitors (H-7, U0126, and PD-98059, respectively). Moreover, the increase in glycerol release induced by widdrol was almost completely blocked by PKC, MEK, and ERK inhibitors. These results suggest that widdrol induces lipolysis through activation of the PKC-MEK-ERK pathway.

Apple AirPods Pro as a Hearing Assistive Device in Patients with Mild to Moderate Hearing Loss.

PURPOSE: This study aimed to assess the feasibility of the Apple AirPods Pro with the headphone accommodation feature as a hearing assistive device for patients with mild to moderate hearing loss (HL). MATERIALS AND METHODS: The study included a total of 35 participants with mild to moderate HL. To determine the degree of HL in the participants, a screening test using pure-tone audiometry was conducted prior to the main tests of functional gain, word recognition score (WRS), and sentence recognition in noisy environments. The study employed two hearing devices: the Bean (a personal sound amplification product, PSAP) and the AirPods Pro. RESULTS: Regarding functional gain, there were no significant differences between the Bean and the AirPods Pro at all frequencies, except 8 kHz. In terms of WRS, both the Bean and the AirPods Pro had higher scores than the unaided condition. In sentence recognition, both the Bean and the AirPods Pro had higher scores than the unaided condition. During real-ear measurement, the Bean demonstrated consistent frequency responses, while the AirPods had a deviation exceeding 10 dB SPL at 6 kHz in the left ear. This deviation was absent for all other frequencies. CONCLUSION: This study shows that the Apple AirPods Pro, with its headphone accommodation feature, performed similarly to a validated PSAP and improved hearing compared to unaided conditions.

Erratum: Cedrol, a Sesquiterpene Isolated from Juniperus chinensis, Inhibits Human Colorectal Tumor Growth associated through Downregulation of Minichromosome Maintenance Proteins.

[This corrects the article on p. 221 in vol. 27, PMID: 36713942.].

Widdrol activates DNA damage checkpoint through the signaling Chk2-p53-Cdc25A-p21-MCM4 pathway in HT29 cells.

Widdrol is an odorant compound isolated from Juniperus chinensis. We previously reported that widdrol induces Gap 1 (G1) phase cell cycle arrest and leads to apoptosis in human colon adenocarcinoma HT29 cells. It was also reported that this cell cycle arrest is associated with the induction of checkpoint kinase 2 (Chk2), p53 phosphorylation and cyclin dependent kinase (Cdk) inhibitor p21 expression. In this paper, we investigated the molecular mechanisms of widdrol on the activation of G1 DNA damage checkpoint at early phase when DNA damages occurred in HT29 cells. First of all, we examined that widdrol breaks DNA directly or not. As the results of DNA electrophoresis and formation of phosphorylated histone H2AX (γH2AX) foci in HT29 cells, widdrol generates DNA double-strand breaks directly within 0.5 h both in vitro and in vivo. Based on this result, the change of proteins related in checkpoint pathway was examined over a time course of 0.5-24 h. Treatment of HT29 cells with widdrol elicits the following: (1) phosphorylation of Chk2 and p53, (2) reduction of cell division cycle 25A (Cdc25A) expression, (3) increase of Cdk inhibitor p21 expression, and (4) decrease of the levels of Cdk2 and cyclin E expression in a time-dependent manner. Moreover, only the expression level of mini-chromosome maintenance 4 (MCM4) protein, a subunit of the eukaryotic DNA replicative helicase, is rapidly down-regulated in HT29 cells treated with widdrol over the same time course, but those of the other MCM proteins are unchanged. Overall, our results indicated that widdrol breaks DNA directly in HT29 cells, and this DNA damage results in checkpoint activation via Chk2-p53-Cdc25A-p21-MCM4 pathway and finally cells go to G1-phase cell cycle arrest and apoptosis.

Cedrol, a Sesquiterpene Isolated from Juniperus chinensis, Inhibits Human Colorectal Tumor Growth associated through Downregulation of Minichromosome Maintenance Proteins.

Cedrol, a sesquiterpene alcohol, isolated from Juniperus chinensis has been reported to inhibit minichromosome maintenance (MCM) proteins as cancer biomarkers in human lung cancer in vitro. In the present study, we investigated the anti-cancer activity of cedrol in vitro and in vivo using human colorectal cancer HT29 cells and a human colorectal tumor xenograft model. Cedrol inhibited MCM protein expression and cell growth in HT29 cells, which are associated with G1 arrest and the induction of apoptosis. We demonstrated that cedrol effectively reduced HT29 tumor growth without apparent weight loss in a human tumor xenograft model. Compared with vehicle- and adriamycin-treated tumor tissues, cedrol induced changes in the tumor tissue structure, resulting in a reduced cell density within the tumor parenchyma and reduced vascularization. Moreover, the expression of MCM7, an important subunit of MCM helicase, was significantly suppressed by cedrol in tumor tissue. Collectively, these results suggest that cedrol may act as a potential anti-cancer agent for colorectal cancer by inhibiting MCM protein expression and tumor growth.

Induction of Cell Cycle Arrest, Apoptosis, and Reducing the Expression of MCM Proteins in Human Lung Carcinoma A549 Cells by Cedrol, Isolated from Juniperus chinensis.

Proteins related to DNA replication have been proposed as cancer biomarkers and targets for anticancer agents. Among them, minichromosome maintenance (MCM) proteins, often overexpressed in various cancer cells, are recognized both as notable biomarkers for cancer diagnosis and as targets for cancer treatment. Here, we investigated the activity of cedrol, a single compound isolated from Juniperus chinensis, in reducing the expression of MCM proteins in human lung carcinoma A549 cells. Remarkably, cedrol also strongly inhibited the expression of all other MCM protein family members in A549 cells. Moreover, cedrol treatment reduced cell viability in A549 cells, accompanied by cell cycle arrest at the G1 phase, and enhanced apoptosis. Taken together, this study broadens our understanding of how cedrol executes its anticancer activity while demonstrating that cedrol has potential application in the treatment of human lung cancer as an inhibitor of MCM proteins.

Formaldehyde exposure induces airway inflammation by increasing eosinophil infiltrations through the regulation of reactive oxygen species production.

Formaldehyde (FA) is a well-known cytotoxic irritant to the airways, but the mechanism of airway inflammation due to FA has not been clarified. In the present study, C57BL/6 mice were exposed to two concentrations (5 and 10ppm) of FA for 6h/day, 5days/week, for 2 weeks. The FA-exposed mice had much higher number of CCR3(+) eosinophils than control mice, and showed upregulated gene expression of CC-chemokine receptor-3 (CCR3), eotaxin and intercellular adhesion molecules-1 (ICAM-1) as well as an increased expression of proinflammatory and Th2 cytokines, such as interleukin (IL)-1ß, IL-4 and IL-5. In addition, FA exposure revealed a considerable increase in the serum levels of IgG1, IgG3, IgA and IgE compared to controls. Histopathological analysis of the lung tissues demonstrated eosinophils and mononuclear cell infiltration of the alveolar cell walls and alveolar spaces. Gene expression of thioredoxin (TRX), redox-regulating antioxidant proteins, was markedly suppressed in FA-exposed mice, and thereby intracellular ROS levels were increased along with increased FA concentration. These results were consistent with an increase in the number of CCR3-expressing eosinophils, and indicate that FA-induced ROS was generated from eosinophils recruited to the inflammatory sites of the airways.

Glycolytic and oxidative muscles under acute glucose supplementation differ in their metabolic responses to fatty acyl-CoA synthetase gene suppression / 한국영양학회지

Purpose@#Skeletal muscles display significant heterogeneity in metabolic responses, owing to the composition of metabolically distinct fiber types. Recently, numerous studies have reported that in skeletal muscles, suppression of genes related to fatty acid channeling alters the triacylglycerol (TAG) synthesis and switches the energy substrates. However, such responses may differ, depending on the type of muscle fiber. Hence, we conducted in vitro and animal studies to compare the metabolic responses of different types of skeletal muscle fibers to the deficiency of fatty acyl-CoA synthetase (Acsl)6, one of the main fatty acidactivating enzymes. @*Methods@#Differentiated skeletal myotubes were transfected with selected Acsl6 short interfering RNA (siRNA), and C57BL/6J mice were subjected to siRNA to induce Acsl6 deficiency. TAG accumulation and expression levels of insulin signaling proteins in response to acute glucose supplementation were measured in immortalized cell-based skeletal myotubes, oxidative muscles (OM), and glycolytic muscles (GM) derived from the animals. @*Results@#Under conditions of high glucose supplementation, suppression of the Acsl6 gene resulted in decreased TAG and glycogen synthesis in the C2C12 skeletal myotubes. The expression of Glut4, a glucose transporter, was similarly downregulated. In the animal study, the level of TAG accumulation in OM was higher than levels determined in GM. However, a similar decrease in TAG accumulation was obtained in the two muscle types in response to Acsl6suppression. Moreover, Acsl6 suppression enhanced the phosphorylation of insulin signaling proteins (Foxo-1, mTORc-1) only in GM, while no such changes were observed in OM. In addition, the induction ratio of phosphorylated proteins in response to glucose or Acsl6 suppression was significantly higher in GM than in OM. @*Conclusion@#The results of this study demonstrate that Acsl6 differentially regulates the energy metabolism of skeletal muscles in response to glucose supplementation, thereby indicating that the fiber type or fiber composition of mixed muscles may skew the results of metabolic studies.

Widdrol, a sesquiterpene isolated from Juniperus chinensis, inhibits angiogenesis by targeting vascular endothelial growth factor receptor 2 signaling.

Widdrol is an odorous compound derived from Juniperus chinensis that is widely used in traditional medicine to treat fever, inflammation and cancer. It was previously reported that widdrol has antitumor activity by apoptosis induction in cancer cells in vitro. However, its anti-angiogenic activity remains elusive. In the present study, we investigated the anti­angiogenic activity of widdrol and the molecular mechanisms involved. Widdrol inhibited cell proliferation via G1 phase arrest induction in human umbilical vein endothelial cells (HUVECs) in a dose-dependent manner. Additionally, it was associated with a decreased expression of cyclin-dependent kinase 2 (CDK2) and an increased expression of p21, a CDK inhibitor. Widdrol significantly inhibited the cell migration and tube formation of HUVECs using an in vitro angiogenesis assay. The results showed that widdrol suppressed phosphorylation of vascular endothelial growth factor receptor 2 (VEGFR2) and its downstream proteins, such as AKT, focal adhesion kinase (FAK) and endothelial nitric oxide synthase (eNOS). Moreover, widdrol effectively reduced tumor growth and blood vessel formation in colon tumor xenograft mice. Collectively, these results suggested that widdrol may act as a potential anti-angiogenic agent by inhibiting vessel sprouting and growth, which may have implications for angioprevention.

Radiographic assessment of clinical root-crown ratios of permanent teeth in a healthy Korean population.

PURPOSE: The aim of this study was to determine the absolute value of the root/crown ratio (R/C ratio) using panoramic radiographs (PRGs) in a healthy Korean population. MATERIALS AND METHODS: In total, 99 patient radiographs (of 50 males and 49 females subjects; aged 16 to 24 years old) were examined, and 2,770 teeth were analyzed. Crown lengths and root lengths were measured with modified Lind's measurements using PACS tools by two examiners in two separate sessions two months apart. All data were analyzed using SPSS. The independent t-test was used to assess for gender differences, and the paired t-test was used to compare both arches with a significance level of P<.05. RESULTS: The mean R/C ratios varied from 1.29 to 1.89 (male: 1.28-1.84; females: 1.31-1.94). The highest R/C ratios were recorded for the mandibular canines (1.89), followed by the maxillary canines (1.79). The lowest R/C ratios were recorded for the maxillary second molars (1.31). In comparison with the maxillary teeth (1.29-1.78), the mandibular teeth yielded the higher R/C ratio (1.47-1.89), and this difference was significant in the females (P<.05). The difference between the genders was not statistically significant, except for the maxillary central incisors, mandibular canines and mandibular first premolars. CONCLUSION: These data may enhance the understanding of the clinical R/C ratio as a useful guideline for determining the status of teeth and the ethnic difference.

Widdrol blocks 3T3-L1 preadipocytes growth and differentiation due to inhibition of mitotic clonal expansion.

Adipocyte differentiation is strongly associated with obesity, which causes metabolic disorders. In this study, we investigated the inhibitory effects of widdrol on 3T3- L1 preadipocyte growth and differentiation. Widdrol decreased lipid droplet accumulation and down-regulated adipogenic transcription factors such as C/EBPalpha, C/EBPbeta, and PPARgamma. Widdrol blocked preadipocyte proliferation and differentiation through the inhibition of mitotic clonal expansion, which was accompanied by the failure of degradation of p21, a cyclin-dependent kinase inhibitor. Cell-cycle analysis clearly indicated that widdrol actively induces cell-cycle arrest at the G1-S phage transition, causing cells to remain in the preadipocyte state. Moreover, widdrol increased p21 expression and inhibited Rb phosphorylation in preadipocyte incubated in a hormone medium. Therefore, these findings clearly suggest that widdrol blocks preadipocyte growth and differentiation through the inhibition of mitotic clonal expansion by p21- and Rb-dependent G1 arrest and can be developed as a potent anti-adipogenic agent for reducing obesity.

A short-term clinical study of marginal bone level change around microthreaded and platform-switched implants.

PURPOSE: The marginal bone levels around implants following restoration are used as a reference for evaluating implant success and survival. Two design concepts that can reduce crestal bone resorption are the microthread and platform-switching concepts. The aims of this study were to analyze the placement of microthreaded and platform-switched implants and their short-term survival rate, as well as the level of bone around the implants. METHODS: The subjects of this study were 27 patients (79 implants) undergoing treatment with microthreaded and platform-switched implants between October 2008 and July 2009 in the Dental Hospital of Yonsei University Department of Periodontology. The patients received follow-up care more than 6 months after the final setting of the prosthesis, at which time periapical radiographs were taken. The marginal bone level was measured from the reference point to the lowest observed point of contact between the marginal bone and the fixture. Comparisons were made between radiographs taken at the time of fixture installation and those taken at the follow-up visit. RESULTS: During the study period (average of 11.8 months after fixture installation and 7.4 months after the prosthesis delivery), the short-term survival rate of microthreaded and platform-switched implants was 100% and the marginal bone loss around implants was 0.16±0.08 mm, the latter of which is lower than the previously reported values. CONCLUSIONS: This short-term clinical study has demonstrated the successful survival rates of a microthread and platform-switched implant system, and that this system is associated with reduced marginal bone loss.

Assertive Behavior in Asking Smokers Not to Smoke among Patients with Vascular Diseases

PURPOSE: The purpose of this study was to identify the assertive behavior of asking smokers not to smoke and investigate the factors related to assertive behavior in patients with vascular diseases. METHODS: Participants were 203 adult Korean patients with vascular diseases such as cerebral infarction and myocardial infarction. Data were collected using questionnaires that included the characteristics of secondhand smoke (SHS), secondhand smoke-related variables (Health belief model factors, health promotion model factors) and level of assertive behavior. Descriptive statistics, t-test, ANOVA and multiple regression using SPSS/WIN 18.0 were performed. RESULTS: Participants who never ask smokers not to smoke was 39.9%, whereas participants who always ask was 7.4%. There was a weak positive relationship between assertive behavior and susceptibility to disease (r=.18), severity of disease (r=.19), benefit of assertive behavior to SHS exposure (r=.10), barrier of assertive behavior to SHS exposure (r=.24), and self-rated health (r=.21) respectively. There was a moderate positive relationship between assertive behavior and self-efficacy of assertive behavior to SHS exposure (r=.49). Health belief model factors explained 15.7% variance and health promotion model factors explained 27.0% of assertive behavior. CONCLUSION: The findings of this study suggest that self-efficacy of assertive behavior to SHS exposure is a very important factor. Therefore the development of a program to foster self-efficacy of assertive behavior regarding SHS exposure in patients with vascular diseases is needed.

Widdrol induces cell cycle arrest, associated with MCM down-regulation, in human colon adenocarcinoma cells.

Widdrol, an odorous compound extracted from Juniperus chinensis, has been shown to inhibit the in vitro growth of in human cancer cells. This study was conducted on cultured human colon adenocarcinoma HT29 cells to elucidate the possible mechanisms by which widdrol exerts its anti-proliferative activity, which until now has remained poorly understood. It was found that widdrol induces accumulation of sub-G1 phase and arrests in the G1 phase of the cell cycle. Induction of G1 arrest by widdrol was correlated with induction of Chk2, p53 phosphorylation and CDK inhibitor p21 expression as well as inhibition of cyclin E, cyclin-dependent kinase (CDK2) and retinoblastoma protein (pRB). Moreover, mini-chromosome maintenance (MCM) proteins were markedly down-regulated in HT29 cells treated with widdrol. Altogether, these results show widdrol possesses potential anti-cancer activity against colon adenocarcinoma cells by inhibiting their proliferation and inducing cell cycle G1 arrest.

Effect of Hizikia Fusiforme Water Extracts on Mouse Immune Cell Activation

Hizikia fusiforme (sea weed fusiforme) has long been used for food source in this country. This study was performed to evalute the immunomodulative effects of Hizikia fusiforme (sea weed fusiforme) in mouse, using in vivo experiments. In vivo experiment, different concentration (0, 50, 500 mg/kg B.W.) of Hizikia fusiforme water extracts were orally administrated into mouse every other day for two weeks. The proliferation of mouse splenocytes, the production of three cytokines (IL-1beta, IL-6, TNF-alpha) secreted by activated macrophage. Splenocyte proliferation was enhanced in mouse orally administrated with 50 mg/kg B.W. and 500 mg/kg B.W. concentration compared to that of control group. Especially, the highest proliferation of spleoncyte was seen from the mouse orally administrated at the concentration of 50 mg/kg B.W. Also, the mouse of Hizikia fusiforme water extracts supplementation group in the both concentrations showed enhanced levels of cytokine production by activated peritoneal macrophages compared to those in control group. The highest level of cytokine (IL-1beta, IL-6, TNF-alpha) production was observed at 50 mg/kg B.W. supplementation group with LPS stimulation in all cases.