Hispidulin Inhibits the Vascular Inflammation Triggered by Porphyromonas gingivalis Lipopolysaccharide.

Hispidulin is a natural bioactive flavonoid that has been studied for its potential therapeutic properties, including its anti-inflammatory, antioxidant, and neuroprotective effects. The aim of this study was to explore whether hispidulin could inhibit the endothelial inflammation triggered by Porphyromonas gingivalis (P. gingivalis) lipopolysaccharide (LPS). The adhesion of monocytes to the vascular endothelium was evaluated through in vitro and ex vivo monocyte adhesion assays. We analyzed the migration of monocytes across the endothelial layer using a transmigration assay. The results showed that treatment with hispidulin decreased the P. gingivalis LPS-induced adhesion of monocytes to endothelial cells and their migration by suppressing the P. gingivalis LPS-triggered expression of intercellular adhesion molecule-1 (ICAM-1) through downregulating nuclear factor-қB (NF-қB). In addition, hispidulin inhibited P. gingivalis LPS-induced mitogen-activated protein kinases (MAPKs) and AKT in endothelial cells. Altogether, the results indicate that hispidulin suppresses the vascular inflammation induced by P. gingivalis LPS. Mechanistically, it prevents the adhesion of monocytes to the vascular endothelium and migration and inhibits NF-қB, MAPKs, and AKT signaling in endothelial cells.

Pentraxin-3 Modulates Osteogenic/Odontogenic Differentiation and Migration of Human Dental Pulp Stem Cells.

Pentraxin-3 (PTX3) is recognized as a modulator of inflammation and a mediator of tissue repair. In this study, we characterized the role of PTX3 on some biological functions of human dental pulp stem cells (HDPSCs). The expression level of PTX3 significantly increased during osteogenic/odontogenic differentiation of HDPSCs, whereas the knockdown of PTX3 decreased this differentiation. Silencing of PTX3 in HDPSCs inhibited their migration and C-X-C chemokine receptor type 4 (CXCR4) expression. Our present study indicates that PTX3 is involved in osteogenic/odontogenic differentiation and migration of HDPSCs, and may contribute to the therapeutic potential of HDPSCs for regeneration and repair.

Involvement of Gastrin-Releasing Peptide Receptor in the Regulation of Adipocyte Differentiation in 3T3-L1 Cells.

Gastrin-releasing peptide (GRP), a member of bombesin-like peptides, and its receptor (GRP-R) play an important role in various physiological and pathological conditions. In this work, we investigated the role of GRP-R on adipogenesis in 3T3-L1 adipocytes. The expression of GRP-R was significantly increased during the adipocyte differentiation of 3T3-L1 cells. The inhibition of GRP-R by the antagonist RC-3095 affected adipogenesis in 3T3-L1 cells, which reduced lipid accumulation and regulated the expression of adipogenic genes. Moreover, cyclic AMP response element-binding protein (CREB) directly bound to the GRP-R promoter upon exposure to adipogenic stimuli. The down-regulation of GRP-R by the knockdown of CREB inhibited adipocyte differentiation of 3T3-L1 cells. Together these results suggest that the regulation of GRP-R activity or expression has an influence on adipogenesis through regulating adipogenic related genes.

The effects of shoulder stabilization exercises and pectoralis minor stretching on balance and maximal shoulder muscle strength of healthy young adults with round shoulder posture.

[Purpose] The purpose of this study was to analyze the effects of pectoralis minor stretching and shoulder strengthening with an elastic band on balance and maximal shoulder muscle strength in young adults with rounded shoulder posture. [Subjects and Methods] Nineteen subjects with rounded shoulder posture were randomly divided into 2 groups: a shoulder stabilization exercise group and a stretching exercise group. The groups performed each exercise for 40 minutes, 3 times a week, for 4 weeks. Static balance (eyes open and closed), dynamic balance (the limits of stability in 4 directions) and shoulder muscle strength in 5 directions were measure before and after the exercises. [Results] The stretching exercise demonstrated a significant difference between the pre- and post-exercise in the static balance with eyes closed and extension and horizontal abduction strength while the stabilization exercise demonstrated significant difference in the left and right directions between the pre- and post-exercise of the dynamic balance and flexion strength. The stabilization exercise demonstrated significant differences shown in the flexion between the pre- and post-test. [Conclusion] The shoulder stabilization and stretching exercises improved the static balance, dynamic balance, and muscle strength.

Gastrin-releasing peptide induces monocyte adhesion to vascular endothelium by upregulating endothelial adhesion molecules.

Gastrin-releasing peptide (GRP) is a neuropeptide that plays roles in various pathophysiological conditions including inflammatory diseases in peripheral tissues; however, little is known about whether GRP can directly regulate endothelial inflammatory processes. In this study, we showed that GRP promotes the adhesion of leukocytes to human umbilical vein endothelial cells (HUVECs) and the aortic endothelium. GRP increased the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) by activating nuclear factor-κB (NF-κB) in endothelial cells. In addition, GRP activated extracellular signal-regulated kinase 1/2 (ERK1/2), p38MAPK, and AKT, and the inhibition of these signaling pathways significantly reduced GRP-induced monocyte adhesion to the endothelium. Overall, our results suggested that GRP may cause endothelial dysfunction, which could be of particular relevance in the development of vascular inflammatory disorders.

Kefir prevented excess fat accumulation in diet-induced obese mice.

Excessive body fat accumulation can result in obesity, which is a serious health concern. Kefir, a probiotic, has recently shown possible health benefits in fighting obesity. This study investigated the inhibitory effects of 0.1 and 0.2% kefir powder on fat accumulation in adipose and liver tissues of high-fat diet (HFD)-induced obese mice. Kefir reduced body weight and epididymal fat pad weight and decreased adipocyte diameters in HFD-induced obese mice. This was supported by decreased expression of genes related to adipogenesis and lipogenesis as well as reduced proinflammatory marker levels in epididymal fat. Along with reduced hepatic triacylglycerol concentrations and serum alanine transaminase and aspartate transaminase activities, genes related to lipogenesis and fatty acid oxidation were downregulated and upregulated, respectively, in liver tissue. Kefir also decreased serum triacylglycerol, total cholesterol, and low-density lipoprotein-cholesterol concentrations. Overall, kefir has the potential to prevent obesity.

The effects of open and closed kinetic chain exercises on the static and dynamic balance of the ankle joints in young healthy women.

[Purpose] The purpose of the present study was to analyze the effects of open kinetic chain and closed kinetic chain exercises on the static and dynamic balance of ankle joints in young healthy women. [Subjects and Methods] Twenty women in their 20s were randomly assigned to two groups of ten women each: an open kinetic chain exercise group and a closed kinetic chain exercise group. Each group performed five sets of exercises three times per week for four weeks. Exercise intensity was increased once after two weeks. The subjects' Romberg's test results and their limits of stability were measured to evaluate their static and dynamic balance. The data were analyzed using a two-way repeated measures analysis of variance test. [Results] In the results of Romberg's test, the main effect of the time showed a significant difference in the trace length with eyes closed (Effect size: d=0.97). In the result of limits of stability, the interaction effect showed a significant difference in the backward, and the main effect of the group showed a significant difference in the forward. [Conclusion] The open kinetic chain and closed kinetic chain exercises both improved the balance of the subjects. The closed kinetic chain exercise was more effective at improving the dynamic balance of young healthy women than the open kinetic chain exercise.

The effect of shoe type on static and dynamic balance during treadmill walking in young healthy women.

[Purpose] The purpose of this study was to analyze the effect of various shoes on the static and dynamic balance of young women in their 20s. [Subjects and Methods] The subjects of the study were 15 healthy young women and repeated measured design. The subjects walked on the treadmill at a speed of 4 km/h for 30 minutes wearing three types of shoes: sneaker, rain boots, and combat boots. Balance was measured by a Romberg test and a limits of stability test. One-way ANOVA was used for statistical analysis. [Results] As the results of the Romberg test, the main effect of time was shown in the EO-COG area, EO-COG length, and EO-COG velocity. As the results of the limits of stability test, the main effects of time in LT, RT, FW, and total. There were significant differences in the LT in the sneaker group, the rain boots group, and the LT and RT in the combat boots group between the pre- and post-test. [Conclusion] The characteristics of shoes such as the materials, hardness, and thickness of the soles, the coefficient of friction of the outsoles, and the collar height affected the static and dynamic balance.

Effects of open and closed kinetic-chain exercises on the muscle strength and muscle activity of the ankle joint in young healthy women.

[Purpose] The purpose of this study was to analyze the effects of open and closed chain exercise on the muscle strength and muscle activity of the ankle joint. [Subjects and Methods] Twenty women in their 20s were randomly assigned to two groups: the open kinetic-chain group and the closed kinetic-chain group. Each group performed 5 sets 3 times per week for 4 weeks. Exercise intensity was increased once after two weeks. The muscle activity of the tibialis anterior, gastrocnemius, tibialis posterior, and peroneus longus muscles were measured. The collected data were analyzed with two-way repeated measures ANOVA. [Results] In the results for muscle strength, both groups showed significant differences in dorsiflexion and plantar flexion between the pre-test and post-test. In the results for muscle activity, no significant differences were noted for either group. [Conclusion] Open and closed kinetic-chain exercises can help to improve muscle strength.

Metabolic engineering of Agrobacterium sp. ATCC31749 for curdlan production from cellobiose.

Curdlan is a commercial polysaccharide made by fermentation of Agrobacterium sp. Its anticipated expansion to larger volume markets demands improvement in its production efficiency. Metabolic engineering for strain improvement has so far been limited due to the lack of genetic tools. This research aimed to identify strong promoters and to engineer a strain that converts cellobiose efficiently to curdlan. Three strong promoters were identified and were used to install an energy-efficient cellobiose phosphorolysis mechanism in a curdlan-producing strain. The engineered strains were shown with enhanced ability to utilize cellobiose, resulting in a 2.5-fold increase in titer. The availability of metabolically engineered strain capable of producing ß-glucan from cellobiose paves the way for its production from cellulose. The identified native promoters from Agrobacterium open up opportunities for further metabolic engineering for improved production of curdlan and other products. The success shown here marks the first such metabolic engineering effort in this microbe.

CCCTC-binding factor controls the homeostatic maintenance and migration of Langerhans cells.

BACKGROUND: Langerhans cells (LCs) are skin-resident dendritic cells (DCs) that orchestrate skin immunity. CCCTC-binding factor (CTCF) is a highly conserved DNA-binding protein that regulates higher-order chromatin organization and is involved in various gene regulation processes. OBJECTIVE: We sought to clarify a possible role for CTCF in LC homeostasis and function in vivo. METHODS: We used a conditional gene deletion mouse system to generate DC- and LC-specific CTCF-ablated mice. Short hairpin RNA-mediated RNA interference was used to silence CTCF expression in human monocyte-derived Langerhans cells. DC populations were assessed by using flow cytometry and immunofluorescence. Gene expression arrays were performed to identify genes regulated by CTCF in LCs. Contact hypersensitivity and epicutaneous sensitization responses were measured to examine the functional significance of CTCF ablation. RESULTS: DC-specific CTCF deletion led to a reduced pool of systemic DCs, with LCs most severely affected. Decreases in epidermal LC numbers were specifically associated with self-turnover defects. Interestingly, CTCF-deficient LCs demonstrated impaired migration out of the epidermis. Whole-transcriptome analyses revealed that genes that promoted cell adhesion were highly expressed, but CCR7 was downregulated in CTCF-depleted LCs. Hapten-induced contact hypersensitivity responses were more sustained in LC-specific CTCF-deficient mice, whereas epicutaneous sensitization to protein antigen was attenuated, indicating that CTCF-dependent LC homeostasis is required for optimal immune function of LCs in a context-dependent manner. CONCLUSION: Our results show that CTCF positively regulates the homeostatic pool and the efficient emigration of LCs, which are required for modulating the functional immune network of the skin.

Involvement of Heme Oxygenase-1 in Orexin-A-induced Angiogenesis in Vascular Endothelial Cells.

The cytoprotective enzyme heme oxygenase-1 (HO-1) influences endothelial cell survival, proliferation, inflammatory response, and angiogenesis in response to various angiogenic stimuli. In this study, we investigate the involvement of HO-1 in the angiogenic activity of orexin-A. We showed that orexin-A stimulates expression and activity of HO-1 in human umbilical vein endothelial cells (HUVECs). Furthermore, we showed that inhibition of HO-1 by tin (Sn) protoporphryin-IX (SnPP) reduced orexin-A-induced angiogenesis in vivo and ex vivo. Orexin-A-stimulated endothelial tube formation and chemotactic activity were also blocked in SnPP-treated vascular endothelial cells. Orexin-A treatment increased the expression of nuclear factor erythroid-derived 2 related factor 2 (Nrf2), and antioxidant response element (ARE) luciferase activity, leading to induction of HO-1. Collectively, these findings indicate that HO-1 plays a role as an important mediator of orexin-A-induced angiogenesis, and provide new possibilities for therapeutic approaches in pathophysiological conditions associated with angiogenesis.

Effect of isotonic and isokinetic exercise on muscle activity and balance of the ankle joint.

[Purpose] This study was performed to examine how the balance of lower limbs and the muscle activities of the tibialis anterior (TA), the medial gastrocnemius (GCM), and the peroneus longus (PL) are influenced by isotonic and isokinetic exercise of the ankle joint. [Subjects] The subjects of this study were healthy adults (n=20), and they were divided into two groups (isotonic=10, isokinetic=10). [Methods] Isotonic group performed 3 sets of 10 contractions at 50% of MVIC and Isokinetic group performed 3 sets of 60°/sec. Muscle activity was measured by EMG and balance was measured by one-leg standing test. [Results] For muscle activity, a main effect of group was found in the non-dominant TA, and the dominant TA, GCM and PL. For balance, a main effect of time was found in both groups for the sway area measured support was provided by the non-dominant side. [Conclusion] In terms of muscle activity, the two groups showed a significant difference, and the isokinetic group showed higher muscle activities. In terms of balance, there was a significant difference between the pre-test and the post-test. The results of this study may help in the selection of exercises for physical therapy, because they show that muscle activity and balance vary according to the type of exercise.

The effects of the length of rain boots on balance during treadmill walking.

[Purpose] Effects of muscle fatigue on lower-extremity balance were evaluated in 12 healthy young women in their 20s while they walked on a treadmill wearing rain boots of different lengths. [Methods] The rain boots were divided into three groups based on the shaft length (Long, Middle, Short). Romberg's test was applied and limits of stability were measured before and after treadmill walking. [Results] Romberg's test showed a significant main effect for time. There were significant differences between the center of gravity area, length, and velocity when the eyes were open and the center of gravity length, velocity, and length/cm(2) when the eyes were closed. Changes in the limits of stability also showed a significant main effect of time. There were significant differences in pre-test and post-test values in the left, right, forward, and total directions. [Conclusion] It was found that muscle fatigue in the lower extremities generated by walking in rain boots affected the joints and the adjuster muscles, depending on shaft lengths. Compensation due to visual feedback and the length of the boot shaft affected movement of the distal joints, resulting in a reduced ability to balance.

Effects of shoe type on lower extremity muscle activity during treadmill walking.

[Purpose] The purpose of this study was to analyze the effects of different shoe types on lower extremity muscle activity in healthy young women by using electromyography. [Subjects and Methods] Fifteen healthy young women in their 20s were included in this single-group repeated measures study. The subjects were divided into three groups: Converse sneakers, rain boots, and combat boots. The subjects walked on a treadmill at 4 km/h for 30 min, during which six muscles were examined using electromyography: the rectus femoris, vastus medialis, semimembranosus, tibialis anterior, peroneus longus, and medial head of the gastrocnemius. Between switching shoe types, a 24-h rest period was instated to prevent the fatigue effect from treadmill walking. [Results] One-way analysis of variance used to compare electromyography results among the three groups showed that the main effect of group differed significantly for the vastus medialis. Vastus medialis activity was higher in the rain boots group than the Converse sneakers group, and it was higher in the combat boots group than rain boots group. [Conclusion] Shoe type affects lower extremity muscle activity. Our findings may help individuals choose the ideal shoes for daily walking.

Effects of different types of exercise on muscle activity and balance control.

[Purpose] This study analyzed the effects of isotonic, isokinetic, and isometric exercises of ankle joint muscles on lower extremity muscle activity and balance control. [Subjects and Methods] The subjects were 30 healthy adults (15 males) in their 20s who were randomly assigned to three different exercise method groups of 10 people each. The isokinetic exercise group performed three sets at an angular velocity of 60°/sec, including a single rest period after every set of 10 repetitions. The isometric exercise group performed three sets consisting of three 15 repetitions of a 15-second exercise followed by a 5-second rest. [Results] Multivariate analysis of variance revealed that depending on the exercise method, the non-dominant tibialis anterior, gastrocnemius muscle, and peroneus longus showed significant differences in muscle activity for weight-bearing non-dominant sides; when the dominant side was weight-bearing, the dominant gastrocnemius and peroneus longus showed significant differences in muscle activity; and the non-dominant and dominant sides showed significant differences in balance control depending on the duration of support in the area. [Conclusion] Muscle fatigue from the three exercise methods produced a decline in muscle activity and balance control; due to the fatigue before exercise, the side that did not perform the exercises was affected.

The effect of olfactory stimuli on the balance ability of stroke patients.

[Purpose] The present study attempted to identify the effect of olfactory stimulation on the balance ability of stroke patients. [Subjects] Thirty-three (33 males) stroke patients participated in the study. The stroke patients were divided into three groups: a black pepper oil (BPO) group (n=11), lavender oil (LVO) group (n=11), and distilled water (DW) group (n=11). [Methods] Two sessions (control trial/stimulus trial) of Romberg's test (eyes open 1 min/eyes closed 1 min) were conducted on a force platform to measure the data for the COP (center of pressure). Olfactory stimulation was provided at as a stimulus. [Results] With the eyes open, a statistically significant difference was found in average anterior posterior displacement (Ymean) and average medial lateral displacement (Xmean) among the three groups when comparing the groups before and after stimulation. The comparison between the eyes open and eyes closed conditions in each group showed a significant difference in the area of the 95% confidence ellipse (area) and Xmean of the BPO group and in the area of the LVO group (area, Xmean). [Conclusion] The findings indicate that the interaction of brain areas activated by the olfactory stimulation exerts an influence on the balance ability of stroke patients.

Glycyrrhetinic acid inhibits Porphyromonas gingivalis lipopolysaccharide-induced vascular permeability via the suppression of interleukin-8.

OBJECTIVE: Porphyromonas gingivalis is a major periodontopathogen that plays a role in the pathogenesis of periodontal disease. In this study, we investigated the effect of 18alpha-glycyrrhetinic acid (18α-GA), a natural triterpenoid compound derived from licorice root extract, on P. gingivalis lipopolysaccharide (LPS)-induced vascular permeability, which is a hallmark of inflammatory diseases such as periodontitis. METHODS: The inhibitory effects of 18α-GA on endothelial permeability were determined by measuring in vivo and in vitro endothelial permeability. Endothelial cells were pretreated with 18α-GA before exposure to P. gingivalis LPS, and total RNA or proteins were extracted and analyzed by reverse transcription polymerase chain reaction or western blotting. RESULTS: Porphyromonas gingivalis LPS-induced endothelial permeability was significantly inhibited by 18α-GA both in vivo and in vitro. 18α-GA reduces P. gingivalis LPS-induced gap formation of endothelial cells. Importantly, 18α-GA modulated the expression and secretion of interleukin-8 (IL-8), a key inducer of vascular permeability, by downregulating nuclear factor-κB (NF-κB). 18α-GA suppressed P. gingivalis LPS-stimulated inhibitor of kappa B (IκB) kinase activation, IκBα phosphorylation, and nuclear translocation of NF-κB. CONCLUSIONS: Overall, these findings suggest that 18α-GA significantly reduces P. gingivalis LPS-induced vascular permeability by repressing NF-κB-dependent endothelial IL-8 production, suggesting its therapeutic potential in P. gingivalis-related vascular diseases.

Kefir inhibits 3T3-L1 adipocyte differentiation through down-regulation of adipogenic transcription factor expression.

BACKGROUND: Kefir, a traditional fermented milk composed of microbial symbionts, is reported to have various health benefits such as anti-tumour, anti-inflammatory, anti-neoplastic and pro-digestive effects. In this study, to elucidate the effects of kefir on adipocyte differentiation and lipid accumulation, three fractions were prepared from kefir culture broth. The inhibitory effects of kefir liquid culture broth fraction (Fr-1), soluble fraction (Fr-2) and insoluble fraction (Fr-3), prepared by sonication of kefir solid culture broth, on adipocyte differentiation in 3T3-L1 preadipocytes were examined. RESULTS: Fr-3 (0.1 mg mL(-1)) significantly decreased lipid accumulation and glycerol-3-phosphate dehydrogenase (GPDH) activity by 60 and 68% respectively without affecting cell viability. In addition, Fr-3 treatment down-regulated the mRNA expression of adipogenic transcription factors including C/EBPα (32%), PPARγ (46%) and SREBP-1c (34%) during adipocyte differentiation compared with untreated control cells. The mRNA expression of adipocyte-specific genes (aP2, FAS and ACC) was also clearly decreased. CONCLUSION: The results suggest that the insoluble fraction of kefir (Fr-3) mediates anti-adipogenic effects through the inhibition of adipocyte differentiation, partly via suppression of the C/EBPα-, SREBP-1c- and PPARγ-dependent pathways.

Inhibition of the Semaphorin 4D-Plexin-B1 axis prevents calcification in vascular smooth muscle cells.

Vascular calcification is common in cardiovascular diseases including atherosclerosis, and is associated with an increased risk of pathological events and mortality. Some semaphorin family members play an important role in atherosclerosis. In the present study, we show that Semaphorin 4D/Sema4D and its Plexin-B1 receptor were significantly upregulated in calcified aorta of a rat chronic kidney disease model. Significantly higher Sema4D and Plexin-B1 expression was also observed during inorganic phosphate-induced calcification of vascular smooth muscle cells. Knockdown of Sema4D or Plexin-B1 genes attenuated both the phosphate-induced osteogenic phenotype of vascular smooth muscle cells, through regulation of SMAD1/5 signaling, as well as apoptosis of vascular smooth muscle cells, through modulation of the Gas6/Axl/Akt survival pathway. Taken together, our results offer new insights on the role of Sema4D and Plexin-B1 as potential therapeutic targets against vascular calcification. [BMB Reports 2023; 56(3): 160-165].