Identification of Reg3ß-producing cells using IL-22-stimulated enteroids.

Reg3ß, a lectin, displays antibacterial activity. This study investigated Reg3ß-expressing cells using IL-22-stimulated enteroids. IL-22 stimulation elevated the mRNA and protein levels of Reg3ß. IL-22 also increased the mRNA levels of CD133 (a transit-amplifying cell marker) and lysozyme (a Paneth cell marker). Immunohistochemistry showed partial colocalization of Reg3ß- and lysozyme-positive cells, suggesting that Paneth cells are one of Reg3ß-producing cells.

The Effect of Methanolic Valeriana officinalis Root Extract on Adipocyte Differentiation and Adiponectin Production in 3T3-L1 Adipocytes.

Adipose tissue is an endocrine organ and its endocrine function is closely associated with type 2 diabetes mellitus. Valeriana officinalis (Valerian) exerts some physiological effects; however, its influence on adipocytes remains unclear. We investigated the effect of methanolic Valerian root extract (Vale) on 3T3-L1 adipocytes. Vale (1, 10, and 100 µg/mL) dose-dependently promoted adipocyte differentiation with increasing lipid accumulation. In addition, Vale significantly increased the mRNA levels in genes associated with adipocyte differentiation, including peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer-binding protein α , and adipocyte protein 2, in dose-dependent manner. Vale also significantly enhanced mRNA and protein levels in adiponectin. A PPARγ antagonist assay and a PPARγ binding assay revealed that Vale-induced increased adipocyte differentiation and adiponectin production were partly associated with direct binding to PPARγ. Valerenic acid, a characteristic component in Valerian, also demonstrated the ability to induce adipocyte differentiation and adiponectin secretion, suggesting that it is one of the functional components in Vale.

Antidiabetic-Like Effects of Naringenin-7-O-glucoside from Edible Chrysanthemum 'Kotobuki' and Naringenin by Activation of the PI3K/Akt Pathway and PPARγ.

Obesity is directly associated with cancer, cardiovascular injury, hypertension, and type 2 diabetes. To date, Yamamoto identified that hot water extracts of edible Chrysanthemum (EC) induced cell size reduction, up-regulation of adiponectin expression, and glucose absorption inhibition in 3T3-L1 cells during adipocyte differentiation. Furthermore, EC showed antidiabetic effects such as improvement in insulin resistance and the down-regulation of the blood glucose level and liver lipid content in type 2 diabetes model mice. In this study, we attempted to identify the antidiabetic components in EC. The methanol fraction from EC that showed relatively strong biological activity was purified by chromatography to obtain acacetin-7-O-glucoside, apigenin-7-O-glucoside, kaempferol-7-O-glucoside, and naringenin-7-O-glucoside. Among the isolated compounds and their aglycones, naringenin (NA) and naringenin-7-O-glucoside (NAG) up-regulated the intracellular accumulation of lipid and adiponectin-secretion and down-regulated the diameter of 3T3-L1 cells during adipocyte differentiation. Because the PPARγ antagonist BADGE and PI3K/Akt inhibitors wortmannin and LY29004 inhibited the intracellular lipid accumulation by NA and NAG associated with adipogenesis, it was considered that NA and NAG showed the above-mentioned activities via the activation of PPARγ as well as phosphorylation of the PI3K/Akt pathway.

Effect of essential amino acids on enteroids: Methionine deprivation suppresses proliferation and affects differentiation in enteroid stem cells.

We investigated the effects of essential amino acids on intestinal stem cell proliferation and differentiation using murine small intestinal organoids (enteroids) from the jejunum. By selectively removing individual essential amino acids from culture medium, we found that 24 h of methionine (Met) deprivation markedly suppressed cell proliferation in enteroids. This effect was rescued when enteroids cultured in Met deprivation media for 12 h were transferred to complete medium, suggesting that Met plays an important role in enteroid cell proliferation. In addition, mRNA levels of the stem cell marker leucine-rich repeat-containing G protein-coupled receptor 5 (Lgr5) decreased in enteroids grown in Met deprivation conditions. Consistent with this observation, Met deprivation also attenuated Lgr5-EGFP fluorescence intensity in enteroids. In contrast, Met deprivation enhanced mRNA levels of the enteroendocrine cell marker chromogranin A (ChgA) and markers of K cells, enterochromaffin cells, goblet cells, and Paneth cells. Immunofluorescence experiments demonstrated that Met deprivation led to an increase in the number of ChgA-positive cells. These results suggest that Met deprivation suppresses stem cell proliferation, thereby promoting differentiation. In conclusion, Met is an important nutrient in the maintenance of intestinal stem cells and Met deprivation potentially affects cell differentiation.

Manual laryngeal fixation facilitates tracheal intubation during chest compression: A randomized crossover manikin study.

PURPOSE: We compared the effectiveness of external manual laryngeal fixation (MLF) for tracheal intubation during chest compression using three laryngoscopes, the Macintosh laryngoscope (McL), McGRATH® MAC (McGRGTH), and Pentax-AWS Airwayscope® (AWS) on an adult manikin. METHODS: Sixteen novice doctors and 15 experienced anesthesiologists performed tracheal intubation during chest compression on an adult manikin using the McL, McGRATH, and AWS with or without MLF. Tracheal intubation time and intubation success rate were measured. RESULTS: In the AWS trial, all novice and experienced doctors successfully secured the airway with or without MLF during chest compression. In McL and McGRATH trials, MLF significantly improved the rate of successful intubation during chest compression compared to without MLF for novice doctors. While intubation time did not significantly differ with or without MLF in the AWS trial, MLF significantly shortened intubation time in McL and McGRATH trials for both novice and experienced doctors. CONCLUSION: These findings suggest that MLF facilitates tracheal intubation with the McL and McGRATH during chest compression.

Evaluation of gum-elastic bougie combined with direct and indirect laryngoscopes in vomitus setting: A randomized simulation trial.

PURPOSE: Videolaryngoscopes may not be useful in the presence of vomitus due to blurred images on the monitor. The objective of our study is to compare the utility of gum-elastic bougie (GEB) application for tracheal intubation with the Macintosh laryngoscope (McL), which is a direct laryngoscope, with that of the Pentax-AWS Airwayscope® (AWS) and McGRATH® MAC (McGRATH) in simulated vomitus settings. METHODS: Sixteen novice doctors performed tracheal intubation on an adult manikin using McL, AWS, and McGRATH with or without GEB under normal and vomitus simulations. RESULTS: In the normal setting the tracheal intubation was successful with the three laryngoscopes regardless of GEB application. In the vomitus setting, the intubation success rate did not significantly improve using McL, while it did using McGRATH or AWS. In the normal settings, GEB application significantly lengthened the intubation time in all three laryngoscopes. By contrast, in the vomitus settings, GEB application significantly shortened the intubation time in all three laryngoscopes. For the comparison of three laryngoscopes, the intubation time did not differ significantly in normal setting, while it was significantly longer in McG and AWS trials than McL trial. CONCLUSION: The GEB application facilitates the tracheal intubation in the vomitus setting using McGRATH and AWS in adult simulation.

Effect of Cricoid Pressure on the Insertion Efficacy of Six Supraglottic Devices: A Crossover Randomized Simulation Trial.

BACKGROUND: No study has ever compared the efficacy of various types of supraglottic devices (SGDs) for securing the airway under cricoid pressure. OBJECTIVE: This study aimed to evaluate the efficacy of six SGDs, LMA-ProSeal (ProSeal), LMA-Classic (Classic), Laryngeal Tube (LT), LMA-Supreme (Supreme), air-Q (air-Q), and i-gel (i-gel), in airway management under cricoid pressure using a manikin. METHODS: Fifteen novice doctors and 16 experienced doctors used the six SGDs under cricoid or sham pressure on an adult manikin. Insertion time, successful ventilation rate, and subjective insertion difficulty on a visual analogue scale (VAS) were measured. RESULTS: Both novice and experienced doctors had a significantly lower ventilation success rate under cricoid pressure than under sham pressure when using the ProSeal, Classic, and LT, but not when using the other three SGDs. Novice doctors required a significantly longer insertion time under cricoid pressure than under sham pressure with all SGDs. Experienced doctors required a significantly longer insertion time under cricoid pressure than with sham pressure when using the ProSeal, Classic, and LT, but not when using the other three SGDs. Subjective insertion difficulty on VAS was significantly higher under cricoid pressure than under sham pressure with all six SGDs. CONCLUSION: Ventilation success rate under cricoid pressure was significantly lower than under sham pressure when using the ProSeal, Classic, and LT, but not when using the other three SGDs in both novice and experienced doctors.

Transforming growth factor-ß1 induces cholesterol synthesis by increasing HMG-CoA reductase mRNA expression in keratinocytes.

In this study, we investigated the effect of TGF-ß1 on cholesterol synthesis in human keratinocytes. TGF-ß1 increased the level of cholesterol and the mRNA level of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase in human keratinocytes. These results show that TGF-ß1 induces cholesterol synthesis by increasing HMG-CoA reductase mRNA expression in human keratinocytes.

Gum-Elastic Bougie Efficacy for Tracheal Intubation During Continuous Chest Compression in Infants-A Crossover Simulation Trial.

BACKGROUND: Recent guidelines for infant cardiopulmonary resuscitation emphasize that all rescuers should minimize interruption of chest compression, even for endotracheal intubation. OBJECTIVE: We compared the utility of application of a gum-elastic bougie (GEB) plus Miller laryngoscope (Mil) with the Mil alone during chest compression on an infant mannequin. METHODS: Sixteen anesthesiologists with more than 2 years of experience performed tracheal intubation on an infant mannequin using the Mil or Mil plus 6Fr GEB, with or without chest compression. Intubation success rate, intubation time, and subjective difficulty scores of laryngoscopy and tube passage through the glottis were measured. RESULTS: In Mil trials, none of the participants failed without compression, whereas four failed with compression (p = 0.03). In Mil-plus-GEB trials, all participants succeeded regardless of chest compression. Intubation time was significantly longer with chest compression in both Mil and Mil-plus-GEB trials (p < 0.001). The intubation time during chest compression was significantly longer in Mil than in Mil-plus-GEB trials (p < 0.001). Difficulty of operation on a visual analog scale (VAS) for laryngoscopy did not significantly differ between Mil and Mil-plus-GEB trials during chest compression, whereas the VAS for tube passage through the glottis was significantly higher in Mil than in Mil-plus-GEB trials. CONCLUSIONS: GEB use shortened the intubation time and improved the success rate of infant tracheal intubation during chest compression by anesthesiologists in simulations.

[Ultrasound-guided Thoracic Epidural and Spinal Anesthesia for the Management of Patients Undergoing Systemic Neurofibroma Resection].

We report successful anesthetic management of a patient undergoing systemic neurofibroma resection using thoracic epidural and spinal anesthesia. A 40- year-old man (weight 66 kg) with von Recklinghausen's disease and intermittent convulsions was scheduled for large-scale tumor resection on the back and arms. He also had chronic obstructive pulmonary disease and stiff neck We decided not to perform operation under general anesthesia. Before epidural or spinal anesthesia, we confirmed that there were no fibromas -in the subdermal lesion for needle progression by ultraso- nography. Thoracic epidural anesthesia was performed at the T2-3 level with 0.25% ropivacaine 5.0 ml, and spinal anesthesia with 0.5% isobaric bupivacaine 3.0 ml. To calm his anxiety, we continuously adminis- tered dexmedetomidine 0.4-0.8 µg · kg(-1) - hr(-1). The operation was performed uneventfully without se- vere pain. Dexmedetomidine was continued until the next morning. The patient showed no signs of convul- sions or intractable pain postoperatively.

Hydrocellular foam dressing increases the leptin level in wound fluid.

Hydrocellular foam dressing (HCF) absorbs excessive wound fluid, which contains various cytokines and growth factors, and ensures a moist environment to promote wound healing. However, the molecular mechanisms underlying the wound fluid component changes induced by HCF are poorly understood. In the present study, we examined the effect of HCF on wound healing and the associated regulatory mechanisms in relation to variations in cytokine levels in the wound fluid. We created full-thickness wounds on the dorsolateral skin of rats and collected the resulting wound fluid samples. HCF was immersed in a plate containing the wound fluids. HCF was then removed and the excess wound fluid remaining in the plate was examined by cytokine array and enzyme-linked immunosorbent assay. We also used a rat model and human dermal fibroblast cultures to examine the effect of wound fluid component changes during the wound healing process. Upon treatment with HCF, leptin levels were upregulated in the wound fluid. Fibroblast proliferation was enhanced and the effect was suppressed in the presence of leptin antagonist. In our in vivo model, HCF increased wound contraction compared with film dressings and this positive effect of HCF was suppressed by addition of leptin antagonist. Our results suggest that dermal fibroblast proliferation is upregulated by HCF due to increased leptin level at the wound surface, and these effects promote wound healing. We believe that the present study contributes to furthering the understanding of the mechanisms underlying the effects of HCF-induced wound healing.

Effects of Various 5,7-Dihydroxyflavone Analogs on Adipogenesis in 3T3-L1 Cells.

We studied the effects of twelve 5,7-dihydroxyflavone analogs on adipogenesis in 3T3-L1 cells. Among the compounds, luteolin, diosmetin, and chrysoeriol partly inhibited adipogenesis by blocking the accumulation of triacylglycerol in the cells. Conversely, tricetin facilitated triacylglycerol accumulation in the cells. The induction of lipogenesis or lipolysis may depend on the number and bonding position of hydroxyl or methoxy groups on the B ring of 5,7-dihydroxyflavone. The mRNA expression levels of adipogenic and lipogenic genes were suppressed by luteolin treatment in the cells, while the mRNA levels of lipolytic genes were not affected. However, the expression levels of the adipogenic, lipogenic, and lipolytic genes, except for adipocyte protein 2 (aP2), were not affected by the addition of tricetin. Moreover, luteolin suppressed glucose transporter type 4 (GLUT4) gene and protein levels. These results indicate that luteolin decreased triacylglycerol levels in 3T3-L1 cells during adipogenesis through the suppression of adipogenic/lipogenic and GLUT4 genes and GLUT4 protein.

Perfluorooctanoic acid binds to peroxisome proliferator-activated receptor γ and promotes adipocyte differentiation in 3T3-L1 adipocytes.

We examined the effect of perfluorooctanoic acid (PFOA) on adipose cells using 3T3-L1 adipocytes and found that PFOA increased adipocyte differentiation, triglyceride accumulation, and the mRNA level of factors related to adipocyte differentiation. In addition, PFOA bound to peroxisome proliferator-activated receptor γ (PPAR γ). These results suggest that PFOA promotes adipocyte differentiation as a PPAR γ ligand.

Hot water extracts of edible Chrysanthemum morifolium Ramat. exert antidiabetic effects in obese diabetic KK-Ay mice.

In this study, the antidiabetic effects of a hot water extract of edible Chrysanthemum morifolium Ramat. (HW-ECM) were investigated in type 2 diabetic mice. HW-ECM improved blood glucose levels and insulin resistance and increased adiponectin mRNA expression in adipose tissues and protein concentrations in the plasma. Moreover, it increased adipose mRNA and protein expressions of peroxisome proliferator-activated receptor γ (PPARγ), a regulator of adiponectin transcription, and mRNA expression of its downstream target genes. It also reduced the adipose cell size and attenuated the mRNA expression of pro-inflammatory adipocytokines in adipose tissues. These data presumably indicate a hypoglycemic mechanism of HW-ECM, involving increased PPARγ expression, decreased the adipocyte sizes, and suppression of chronic inflammation in adipose tissues. Finally, elevated adiponectin levels lead to amelioration of insulin resistance and the corresponding hypoglycemic effects. Therefore, HW-ECM indicates its potential as a functional food for type 2 diabetes.

Hydrocellular foam dressings promote wound healing associated with decrease in inflammation in rat periwound skin and granulation tissue, compared with hydrocolloid dressings.

The effects of modern dressings on inflammation, which represent the earliest phase of wound healing, are poorly understood. We investigated the effects of modern hydrocellular foam dressings (HCFs) on wound healing and on the gene expression levels of the inflammatory markers--interleukin (IL)-1ß, IL-6, and IL-10--in rat periwound skin and granulation tissue by quantitative reverse transcription-polymerase chain reaction. HCF absorbed significantly higher volume of water than hydrocolloid dressing (HCD) and increased the contraction of wounds. Polymorphonuclear neutrophils were massively infiltrated to the wound edge and boarded between granulation and dermis in the HCD group. IL-1ß, IL-6, and IL-10 mRNA levels were significantly decreased in the periwound skin around the wounds and granulation tissue covered with HCF. These findings suggest that HCF may promote wound healing along with decrease in inflammation by reducing gene expression levels of IL-1ß, IL-6, and IL-10.

Calcitonin gene-related peptide regulates mitogen-activated protein kinase pathway to decrease transforming growth factor ß1-induced hepatic plasminogen activator inhibitor-1 mRNA expression in HepG2 cells.

Transforming growth factor (TGF) ß1-induced plasminogen activator inhibitor (PAI)-1 is one of factors associated with the development of hepatic fibrosis. Calcitonin gene-related peptide (CGRP) shows hepatoprotective effect during hepatic injuries, including fibrosis. However, the effects of CGRP on PAI-1 expression induced by TGFß1 are unknown. In this study, we investigated the effect of CGRP on TGFß1-induced PAI-1 expression and its regulatory mechanisms in HepG2 cells. CGRP inhibited TGFß1-induced PAI-1 expression. H89, a protein kinase A inhibitor, abolished the inhibition of TGFß1-induced PAI-1 expression by CGRP. TGFß1 activated mitogen-activated protein kinase (MAPK), including extracellular signal-regulated kinase, c-jun NH2-terminal kinase, and p38, and this activation was abolished by CGRP. These results show that the CGRP-induced cAMP/PKA activation suppresses activation of MAPK induced by TGFß1, leading to decreased PAI-1 expression in HepG2 cells.

Calcitonin gene-related peptide upregulates serum amyloid A synthesis through activation of interleukin-6.

We found that calcitonin gene-related peptide (CGRP) enhanced the expression of levels of serum amyloid A (SAA) and interleukin-6 (IL-6) in HepG2. In addition, CGRP-induced SAA1/2 mRNA expression was blocked by an anti-IL-6 neutralizing antibody in HepG2. These results suggest that CGRP promotes SAA synthesis through activation of IL-6 in human hepatocytes.

Novel angiotensin I-converting enzyme inhibitory peptides found in a thermolysin-treated elastin with antihypertensive activity.

Angiotensin I-converting enzyme (ACE) inhibitory activity was generated from elastin and collagen by hydrolyzing with thermolysin. The IC50 value of 531.6 µg/mL for ACE inhibition by the elastin hydrolysate was five times less than 2885.1 µg/mL by the collagen hydrolysate. We confirmed the antihypertensive activity of the elastin hydrolysate in vivo by feeding spontaneously hypertensive rats (male) on a diet containing 1% of the elastin hydrolysate for 9 weeks. About 4 week later, the systolic blood pressure of the rats in the elastin hydrolysate group had become significantly lower than that of the control group. We identified novel ACE inhibitory peptides, VGHyp, VVPG and VYPGG, in the elastin hydrolysate by using a protein sequencer and quadrupole linear ion trap (QIT)-LC/MS/MS. VYPGG had the highest IC50 value of 244 µM against ACE and may have potential use as a functional food.

Accelerating effect of soy peptides containing collagen peptides on type I and III collagen levels in rat skin.

Two weeks of feeding soy peptides containing 2% collagen peptides increased the levels of type I and III tropocollagen and their mRNAs. In contrast, the diet did not increase the mRNA levels of rat hyaluronan synthases, serine palmitoyltransferase (the rate-limiting enzyme of ceramide synthesis), and 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (the key enzyme of cholesterol synthesis). These results suggest that feeding of soy peptides with collagen peptides specifically enhanced the tropocollagen level in the skin.