Bojungikgi-tang improves skin barrier function and immune response in atopic dermatitis mice fed a low aryl hydrocarbon receptor ligand diet.

BACKGROUND: The aryl hydrocarbon receptor (AhR) is a transcription factor that plays a crucial role in regulating the immune system and maintaining skin barrier function. AhR signaling is pivotal in the pathogenesis of inflammatory diseases such as atopic dermatitis (AD), and the absence of AhR ligands further contributes to the progression or worsening of AD symptoms. METHODS: AD was induced with 2,4-dinitrochlorobenzene (DNCB), and Bojungikgi-tang (BJIKT) was administered orally daily for 10 weeks. Serum IgE, splenocyte IL-4, and IFN-γ levels, skin barrier genes, and AhR target gene expressions were analyzed using RNA-sequencing analysis. Spleen tissues were extracted for fluorescence-activated cell sorting (FACS) analysis to analyze the effect of BJIKT on immune responses. A correlation analysis was conducted to analyze the correlation between immune markers and skin barrier genes and AhR target genes. RESULTS: BJIKT effectively improved AD symptoms in AD mice fed a low AhR ligand diet by reducing neutrophil and eosinophil counts, lowering IgE levels in the blood, and decreasing IL-4 and IFN-γ levels in the splenocytes. Additionally, BJIKT significantly reduced epithelial skin thickness and transepidermal water loss (TEWL) values and reversed the decreased expression of skin barrier genes. BJIKT also considerably altered the expression of AhR target genes, including Ahr, Ahrr, cytochrome P450 1A1 (CYP1A1), and CYP1B1. Furthermore, AhR target pathway genes were negatively correlated with immune cell subtypes, including CD4 + and CD8 + T cells and macrophages (CD11b + F4/80 +) at the systemic level. CONCLUSIONS: BJIKT can regulate AhR activation and may help reduce inflammation in AD by regulating the expression of skin barrier genes and immune responses.

Metaverse-based social skills training programme for children with autism spectrum disorder to improve social interaction ability: an open-label, single-centre, randomised controlled pilot trial.

Background: Autism spectrum disorder (ASD) is characterised by abnormalities in social interactions and restricted and repetitive behaviors. Children with high-functioning ASD (HFASD), lack social communication skills, do not interact with others, and lack peer relationships. We aimed to develop, and evaluate the feasibility of, a metaverse-based programme to enhance the social skills of children with HFASD. Methods: This open-label, single-centre, pilot parallel randomised controlled trial (RCT) was conducted on boys aged 7-12 years with HFASD. Children were recruited from a treatment centre for children with HFASD in Korea or by self-referral through online community webpages for the parents of children with HFASD. Participants were randomly assigned (1:1) by a blinded researcher to receive either four weeks of a metaverse-based social skills training programme or a control group. Randomisation was stratified by age (children aged 7-9 and 10-12 years) using permuted blocks (block size 4). The metaverse-based social skills training programme was delivered via the metaverse platforms (Roblox) and Zoom. Children in the intervention group completed the metaverse-based social skills training programme at home for four weeks. The intervention consisted of four sessions, one session per week, for 60 min each. The control group did not receive any interventions. The primary outcome measure was the median change in the Social Responsiveness Scale-2 (SRS-2) scores from pre-to post-intervention. SRS-2 is an assessment tool used to confirm the effectiveness of social interactions. Higher scores indicate lower social functioning. The trial is registered with CRIS Registration Number; KCT0006859. Findings: Between February 14, 2022, and March 31, 2022, 20 participants were enrolled. Overall, 15 children (median [Interquartile range (IQR)] age, intervention group: 9.0 [8.0-10.0]; control group: 8.5 [8.0-10.0]) participated in the programme. The intervention group included nine participants (60%), and the control group included six participants (40%). The SRS-2 total scores for the intervention group decreased from baseline 96.0 (IQR: 74.0-112.0) to post-intervention 85.0 (IQR: 84.0-103.0). The group median difference in SRS-2 scores between the intervention and control groups was 11.5 (95% CI: 8.5-14.0), with a further reduction in the intervention group. Similar trends were seen for social cognition (group median difference, 95% CI: 2.0, 1.0-4.0), social communication (group median difference, 95% CI: 2.0, 1.0-4.0), and autistic mannerism (group median difference, 95% CI: 4.0, 1.0-5.0). There were no adverse events related to study participation. Interpretation: The findings of this feasibility study suggest that children with HFASD can potentially be familiarised, through metaverse-based programmes, with real-life social situations to improve sociality and reduce emotional and behavioural problems. Such interventions could be delivered at home and possibly be extended to target groups that have difficulty in interacting with peers offline. Funding: The Institute of Information & Communications Technology Planning & Evaluation grant, via the Ministry of Science and ICT of the South Korean Government.

Primary and additional treatment preference in aggressive retinopathy of prematurity and type 1 retinopathy of prematurity.

OBJECTIVE: This study aimed to evaluate the preference for antivascular endothelial growth factor (anti-VEGF) versus laser ablation therapy as primary and additional treatment in aggressive retinopathy of prematurity (ROP) and type 1 ROP. METHODS: This multicentre retrospective study was conducted at nine medical centres across South Korea. A total of 94 preterm infants with ROP who underwent primary treatment between January 2020 and December 2021 were enrolled. All eyes were classified as having type 1 ROP or aggressive ROP. Data on the zone, primary treatment chosen, injection dose, presence of reactivation and additional treatment were collected and analysed. RESULTS: Seventy infants (131 eyes) with type 1 ROP and 24 infants (45 eyes) with aggressive ROP were included. Anti-VEGF injection was selected as the primary treatment in 74.05% of the infants with type 1 ROP and 88.89% with aggressive ROP. Anti-VEGF injection was selected as the ROP was located in zone I or posterior zone II, and laser ablation was selected when it was located in zone II. The anti-VEGF injection doses varied and tended to be higher in the aggressive ROP group. Infants with aggressive ROP were 2.08 times more likely to require additional treatment than those with type 1 ROP. When ROP reactivation occurred, laser therapy was preferred as an additional treatment. CONCLUSION: In Korea, the preference for anti-VEGF therapy or laser therapy differed according to ROP subtype, zone and primary or secondary treatment. These findings suggest that ROP treatment are considered according to ROP subtype, location and reactivation.

The association between amniotic fluid-derived inflammatory mediators and the risk of retinopathy of prematurity.

Prenatal and perinatal infections and inflammation appear to associated with the development of retinopathy of prematurity (ROP). In this study, we evaluated whether inflammatory mediators in amniotic fluid (AF) retrieved during cesarean delivery influence the development of ROP in very low birth weight (VLBW) infants. This retrospective study included 16 and 32 VLBW infants who did and did not develop any stage of ROP, respectively. Each infant with ROP was matched with 2 infants without ROP based on days of ventilation care, gestational age, and birth weight. AF was obtained during cesarean delivery, and the levels of intra-amniotic inflammatory mediators such as interleukin (IL)-1ß, IL-2, IL-6, IL-8, IL-10, matrix metalloproteinase (MMP)-2, MMP-8, MMP-9, and tumor necrosis factor (TNF)-α were measured using a Human Magnetic Luminex assay (R&D Systems, Minneapolis, MN). The differences in the levels of inflammatory mediators according to the presence or absence of ROP were compared. In patients who developed ROP, the level of MMP-2 in the AF was significantly increased (P = .011), whereas the levels of IL-10 and TNF-α were significantly decreased (P = .028 and .046, respectively) compared with those in infants who did not develop ROP. The levels of the other mediators were not significantly different between the 2 groups. Multivariate regression analysis showed that MMP-2 was a risk factor for the development of ROP (odds ratio, 2.445; 95% confidence interval, 1.170-5.106; P = .017). The concentration of MMP-2 in AF is an independent factor in the development of ROP. Further studies are needed to determine whether the levels of inflammatory mediators in AF affect the ROP severity.

Prognostic factors in fluid-gas exchange after primary idiopathic macular hole surgery.

PURPOSE: This retrospective study aimed to evaluate the prognostic factors associated with the success of fluid-gas exchange in patients who had undergone failed primary idiopathic macular hole (IMH) surgery. METHODS: In total, 19 eyes of 19 patients with failed IMH surgery who then underwent fluid-gas exchange were included. Of those, 18 eyes had macular hole (MH) closure (successful, 15 eyes; unsuccessful, 3 eyes). Demographics, pre-operative characteristics, and pre-procedural characteristics were assessed. The patients were divided into successful (U or V-type closure) and unsuccessful groups (W-type or unclosed), following fluid-gas exchange. One eye was unclosed after fluid-gas exchange; therefore, this patient underwent additional vitrectomy for MH closure (unsuccessful). RESULTS: The outcomes of the fluid-gas exchange were categorized as unclosed or as U-type, V-type, or W-type closure. None of the patients experienced complications after the procedure. The successful group showed a significantly lower pre-operative and pre-procedural minimum diameter, base diameter, and macular hole volume, and higher pre-operative and pre-procedural macular hole index, hole form factor, and tractional hole index values. Moreover, a better visual prognosis was observed in the successful group. CONCLUSION: These findings suggest that indices predicting favorable results of primary surgery for IMH are useful for predicting the success of fluid-gas exchange in patients with failed primary MH surgery.

Effective End-to-End Deep Learning Process in Medical Imaging Using Independent Task Learning: Application for Diagnosis of Maxillary Sinusitis.

PURPOSE: This study aimed to propose an effective end-to-end process in medical imaging using an independent task learning (ITL) algorithm and to evaluate its performance in maxillary sinusitis applications. MATERIALS AND METHODS: For the internal dataset, 2122 Waters' view X-ray images, which included 1376 normal and 746 sinusitis images, were divided into training (n=1824) and test (n=298) datasets. For external validation, 700 images, including 379 normal and 321 sinusitis images, from three different institutions were evaluated. To develop the automatic diagnosis system algorithm, four processing steps were performed: 1) preprocessing for ITL, 2) facial patch detection, 3) maxillary sinusitis detection, and 4) a localization report with the sinusitis detector. RESULTS: The accuracy of facial patch detection, which was the first step in the end-to-end algorithm, was 100%, 100%, 99.5%, and 97.5% for the internal set and external validation sets #1, #2, and #3, respectively. The accuracy and area under the receiver operating characteristic curve (AUC) of maxillary sinusitis detection were 88.93% (0.89), 91.67% (0.90), 90.45% (0.86), and 85.13% (0.85) for the internal set and external validation sets #1, #2, and #3, respectively. The accuracy and AUC of the fully automatic sinusitis diagnosis system, including site localization, were 79.87% (0.80), 84.67% (0.82), 83.92% (0.82), and 73.85% (0.74) for the internal set and external validation sets #1, #2, and #3, respectively. CONCLUSION: ITL application for maxillary sinusitis showed reasonable performance in internal and external validation tests, compared with applications used in previous studies.

Bioactive Lipid O-cyclic phytosphingosine-1-phosphate Promotes Differentiation of Human Embryonic Stem Cells into Cardiomyocytes via ALK3/BMPR Signaling.

Adult human cardiomyocytes have an extremely limited proliferative capacity, which poses a great barrier to regenerative medicine and research. Human embryonic stem cells (hESCs) have been proposed as an alternative source to generate large numbers of clinical grade cardiomyocytes (CMs) that can have potential therapeutic applications to treat cardiac diseases. Previous studies have shown that bioactive lipids are involved in diverse cellular responses including cardiogenesis. In this study, we explored the novel function of the chemically synthesized bioactive lipid O-cyclic phytosphingosine-1-phosphate (cP1P) as an inducer of cardiac differentiation. Here, we identified cP1P as a novel factor that significantly enhances the differentiation potential of hESCs into cardiomyocytes. Treatment with cP1P augments the beating colony number and contracting area of CMs. Furthermore, we elucidated the molecular mechanism of cP1P regulating SMAD1/5/8 signaling via the ALK3/BMP receptor cascade during cardiac differentiation. Our result provides a new insight for cP1P usage to improve the quality of CM differentiation for regenerative therapies.

A new optical coherence tomography method for subfoveal choroidal thickness measurement: A single-center observational study.

ABSTRACT: Several macular pathologies are associated with choroidal vascular supply and thus require choroidal thickness evaluation. However, there is no standard subfoveal choroidal thickness (SFCT) measurement method. This study investigated a new method of SFCT measurement, using optical coherence tomography.This is a single-center observational study. Eighty-two senior medical students participated as observers and independently measured the SFCT on 5 standard optical coherence tomography images of healthy adults in a tertiary care setting. They used 3 different methods: the observer's own method, the conventional method, perpendicular to the retinal pigment epithelium, and the new method, along a virtual line connecting the umbo with the most elevated point of the ellipsoid. Additionally, the SFCT angle-the angle between the measurement line and the vertical line of the image-was measured and compared between methods. The intraclass correlation coefficient was used to determine interpersonal variability.The intraclass correlation coefficients for SFCT, measured using methods 1, 2, and 3, were 0.853, 0.880, and 0.896, respectively (P < .001 for all). Method 3 was the highest. The intraclass correlation coefficients of the SFCT angles were 0.647, 0.842, and 0.307, respectively (P < .001 for all).The new method showed the lowest interpersonal variability and could therefore be a reliable standard for SFCT measurement, even in foveae with a steep slope on optical coherence tomography.Trial registration: Not applicable.

Genotoxicity of Asiasari Radix et Rhizoma (Aristolochiaceae) ethanolic extract in vitro and in vivo.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional herbal medicines have diverse efficacy and are increasingly used worldwide. However, some of these herbal medicines have toxicities or side effects, but the scientific understanding of traditional herbal medicine toxicity has not yet been established. Asiasari Radix et Rhizoma (ARE) is known as a herbal medicine used to relieve pain, and recent studies have shown that ARE has anticancer and antimelanogenesis efficacy. AIM OF THE STUDY: Current study was conducted to assess the potential genotoxicity of an ethanolic extract of ARE. MATERIALS AND METHODS: The genotoxixity of ARE was confirmed by the bacterial reverse mutation assay (Ames test), a mammalian chromosomal aberration test, and a micronucleus test in vivo using ICR mice and comet assay using Sprague-Dawley rats. RESULTS: ARE showed no genotoxicity in a micronucleus test up to 2000 mg/kg body weight in vivo. By contrast, the chromosomal aberration test showed that ARE induced an increase in the number of chromosomal aberrations after treatment for 6 h with a metabolic activation system and for 6 and 22 h without the metabolic activation system when compared with vehicle control. In the Ames test, all strains except TA1535, with or without a metabolic activation system, showed an increase in the number of revertant mutant colonies in the ARE-treated group. In comet assay, DNA damage was observed in the stomach when ARE was administered. CONCLUSION: ARE potentially shows genotoxicity by inducing DNA damage.

Age and Gender Effects on Genotoxicity in Diesel Exhaust Particles Exposed C57BL/6 Mice.

There is growing evidence that the accumulation of DNA damage induced by fine particulate matter (PM2.5) exposure is an underlying mechanism of pulmonary disease onset and progression. However, there is a lack of experimental evidence on whether common factors (age, gender) affect PM2.5 induced genomic damage. Here, we assessed the DNA damage potency of PM2.5 using conventional genotoxicity testing in old male and female mice aged 8 and 40 weeks. Mice were intratracheally instilled with diesel exhaust PM2.5 (DEP, NIST SRM 1650b), twice a week for 4 weeks. Exposure to DEP was not associated with an increase in the frequency of micronucleated polychromatic erythrocytes and did not induce a systemic genotoxic effect in the bone marrow. Meanwhile, the results from the comet assay showed a significant increase in DNA damage in DEP exposed mouse lung specimens. The positive relationship between DEP exposure and DNA damage is stronger in the older than in the younger group. Statistical analysis showed that there was a modifying effect of age on the association between PM2.5 exposure and DNA damage. Our results suggest that the age factor should be considered to better understand the cellular adverse effects of PM2.5.

Determination of Genotoxicity Attributed to Diesel Exhaust Particles in Normal Human Embryonic Lung Cell (WI-38) Line.

Several epidemiological studies concluded that inhalation of diesel exhaust particles (DEP) is associated with an increase in the relative risk of lung cancer. In vitro research evaluating the genetic damage and/or changes in gene expression have been attempted to explain the relationship between DEP exposure and carcinogenicity. However, to date, investigations have been largely confined to studies in immortalized or tumorigenic epithelial cell models. Few studies have investigated damage at the chromosomal level to DEP exposure in normal cell lines. Here, we present the genotoxic effects of DEP in normal cells (embryonic human lung fibroblasts) by conventional genotoxicity testing (micronuclei (MN) and comet assay). We show the differentially expressed genes and enriched pathways in DEP-exposed WI-38 cells using RNA sequencing data. We observed a significant increase in single-strand DNA breaks and the frequency of MN in DEP-exposed cells in a dose-dependent manner. The differentially expressed genes following DEP exposure were significantly enriched in the pathway for responding to xenobiotics and DNA damage. Taken together, these results show that DEP exposure induced DNA damage at the chromosomal level in normal human lung cells and provide information on the expression of genes associated with genotoxic stress.

Sinonasal Rhabdomyosarcoma Metastasis in Bilateral Multiple Extraocular Muscles: A Case Report and Brief Literature Review.

Isolated metastasis in the extraocular muscle (EOM) is uncommon, while metastases in bilateral multiple EOMs is even rarer. Rhabdomyosarcoma (RMS) is a rare soft-tissue malignancy that usually occurs in the pediatric population and is one of the primary malignancies of isolated EOM metastasis. Here, we present a case of sinonasal RMS metastasis to multiple bilateral EOMs along with a brief review of 10 previously reported cases of RMS metastasis in EOMs.

Retinal vascular development in an immature retina at 33-34 weeks postmenstrual age predicts retinopathy of prematurity.

In preterm birth, the immature retina can develop a potentially blinding disorder of the eye known as retinopathy of prematurity (ROP). The vaso-proliferative phase of ROP begins at an approximate postmenstrual age (PMA) of 32 weeks. There is little or no evidence of an association between ROP development and retinal status in the early vaso-proliferative phase. We aimed to evaluate the retinal vascular findings of infants at 33-34 weeks PMA to determine their risk of ROP. We reviewed 130 serial wide-field retinal images from 65 preterm infants born before the gestational age of 31 weeks. ROP occurred more frequently in infants having a leading vascular edge within posterior Zone II. This was in contrast to normal infants, who are characterized by complete retinal vascularization up to Zone II at 34 weeks PMA. The probability of ROP development in preterm infants with retinal edge hemorrhage was 24.58 times higher than in preterm infants without retinal edge hemorrhage. Eyes with ROP that required treatment showed significantly delayed retinal vascularization accompanied by pre-plus disease. In conclusion, retinal status in the early vaso-proliferation phase might determine the risk of ROP.

Curcumin induces expression of 15-hydroxyprostaglandin dehydrogenase in gastric mucosal cells and mouse stomach in vivo: AP-1 as a potential target.

15-Hydroxyprostaglandin dehydrogenase (15-PGDH) catalyzes the conversion of oncogenic prostaglandin E2 to non-tumerigenic 15-keto prostaglandin E2. In the present study, we found that curcumin, a yellow coloring agent present in the rhizome of Curcuma longa Linn (Zingiberaceae), induced expression of 15-PGDH at the both transcriptional and translational levels in normal rat gastric mucosal cells. By using deletion constructs of 15-PGDH promoter, we were able to demonstrate that activator protein-1 (AP-1) is the principal transcription factor responsible for regulating curcumin-induced 15-PGDH expression. Curcumin enhanced the expression of c-Jun and c-Fos that are functional subunits of AP-1, in the nuclear fraction of cells. Silencing of c-Jun suppressed curcumin-induced expression of 15-PGDH. Moreover, the chromatin immunoprecipitation assay revealed curcumin-induced binding of c-Jun to the AP-1 consensus sequence present in the 15-PGDH promoter. Curcumin increased phosphorylation of ERK1/2 and JNK, and pharmacologic inhibition of these kinases abrogated the curcumin-induced phosphorylation of c-Jun and 15-PGDH expression. In contrast, tetrahydrocurcumin which lacks the α,ß-unsaturated carbonyl group failed to induce 15-PGDH expression, suggesting that the electrophilic carbonyl group of curcumin is essential for its induction of 15-PGDH expression. Curcumin restored the expression of 15-PGDH which is down-regulated by Helicobacter pylori through suppression of DNA methyltransferase 1. In addition, oral administration of curcumin increased the expression of 15-PGDH and its regulators such as p-ERK1/2, p-JNK, and c-Jun in the mouse stomach. Taken together, these findings suggest that curcumin-induced upregulation of 15-PGDH may contribute to chemopreventive effects of this phytochemical on inflammation-associated gastric carcinogenesis.

Correlation between macular edema recurrence and macular capillary network destruction in branch retinal vein occlusion.

BACKGROUND: The aim of this study was to evaluate the correlation between changes in the macular capillary network and macular edema (ME) recurrence with branch retinal vein occlusion (BRVO) using swept-source optical coherence tomography angiography (SS-OCTA). METHODS: We reviewed the data for 43 patients with treatment-naïve ME associated with BRVO. Patients who received intravitreal bevacizumab injection were divided into two groups based on ME recurrence at 6 months after edema resolution. The perifoveal capillary morphology and the macular capillary vessel density (VD) were retrospectively analyzed using en face SS-OCTA after ME resolution. RESULTS: The perifoveal capillary ring loss in the superficial capillary plexus (SCP) and deep capillary plexus (DCP) was more common in the ME recurrence group (n = 22) than in the no ME recurrence group (p = 0.047 and p = 0.002). Relative to the findings in the no ME recurrence groups, the destruction of the perifoveal capillary ring was more severe in the DCP (30.0° vs 87.3°, p = 0.001) than in the SCP (17.3° vs 69.5°, p = 0.006) in the ME recurrence group. The hemi-VD disparity between the affected and the unaffected areas in the SCP and DCP showed significant differences (p = 0.031 and p = 0.017), while macular VD showed no differences between the groups. CONCLUSIONS: Destruction of the perifoveal capillary ring and hemi-VD disparity could be related to ME recurrence in BRVO. Therefore, these factors may be helpful in predicting ME recurrence.

Orbital infarction syndrome after cerebral aneurysm surgery: A case series and literature review.

To report the clinical characteristics and retinal abnormalities associated with orbital infarction syndrome after cerebral aneurysm clipping surgery.In this retrospective case series, we evaluated 4 cases of orbital infarction syndrome using fluorescein angiography, optical coherence tomography, and computed tomography images from January 2011 to May 2014. The medical records of these patients including age, sex, laterality of the eyes, visual acuity, intraocular pressure, duration of the operation, location of the aneurysms, and surgical method with the type of approach used to reach the aneurysmal lesions were evaluated.Aneurysms were located in either the anterior or the posterior communicating artery. Two patients had subarachnoid hemorrhage arising from a ruptured aneurysm, whereas 2 other patients had unruptured aneurysms. Clipping was performed by 3 different surgeons using the pterional craniotomy. The mean time interval from aneurysmal clipping to awareness of vision loss was 10.75 ±â€Š13.8 days. In all patients, optic atrophy and irreversible deterioration of visual acuity ensued. Retinal edema, retinal vascular abnormality, or choroidal hypoperfusion was identified in these patients.Orbital infarction syndrome is a rare but devastating complication of brain aneurysm clipping surgery. The associated retinal ischemia is not only due to the involvement of the retinal vessels, but also the choroidal circulation.

Topical Application of A New Herbal Complex, NI-01, Ameliorates House Dust Mite-Induced Atopic Dermatitis in NC/Nga Mice.

Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by pruritus and cutaneous dry skin. Here, we investigated whether topical application of NI-01 composed of six herbal medicines has a therapeutic effect on AD in vivo. Twelve marker compounds of NI-01 were analyzed by high-performance liquid chromatography with a photodiode array detector for quality control. To induce AD, house dust mite extract was applied to the shaved dorsal skin and ear surfaces of NC/Nga mice twice a week for 6 weeks. NI-01 (1, 2, or 4 mg/mouse) was applied daily to the site for experiment periods. The coefficient of determination of each compound showed good linearity (≥ 0.9999). The recovery rate of the 12 marker components was 96.77%-105.17%; intra and interday precision and repeatability were ≤ 1.40%. Topical application of NI-01 reduced house dust mite induced AD symptoms. The increased expressions of interleukin-4 and intercellular adhesion molecule-1 caused by house dust mites were markedly suppressed in NI-01-treated mice. Corticosterone levels significantly decreased, whereas serotonin levels increased with NI-01 application. These results suggest that NI-01 alleviates AD symptoms by inhibiting infiltration of inflammatory cells, thereby decreasing AD-related stress. NI-01 could be beneficial for the treatment of AD-like skin diseases.

The vasodilatory effect of the antidiabetic drug linagliptin via inhibition of Rho-associated protein kinase in aortic smooth muscle.

AIMS: The vasodilatory effects of the anti-diabetic drug, linagliptin in phenylephrine-precontracted aortic rings were investigated. MATERIALS AND METHODS: Male New Zealand White rabbits were used in the experiment and its arterial tone was measured by using myogragh system. KEY FINDINGS: Linagliptin induced vasodilation in a concentration-dependent manner. The vasodilatory effect of linagliptin was not affected by the absence of the endothelium, or by pretreatment with a nitric oxide synthase inhibitor (L-NAME) or a small-conductance Ca2+-activated K+ channel inhibitor (apamin). Moreover, application of the adenylyl cyclase inhibitor SQ22536, protein kinase A (PKA) inhibitor KT5720, guanylyl cyclase inhibitor ODQ, or protein kinase G (PKG) inhibitor KT5823 did not alter the vasodilatory effect of linagliptin. However, inhibition of Rho-associated protein kinase by Y-27632 significantly attenuated linagliptin-induced vasodilation. Ion channel involvement in the vasodilatory effect of linagliptin was also investigated. Pretreatment with the vascular K+ channel inhibitors glibenclamide (ATP-sensitive K+ channels), Ba2+ (inwardly rectifying K+ channels), 4-AP (voltage-dependent K+ channels), and paxilline (large conductance Ca2+-activated K+ channels) did not affect linagliptin-induced vasodilation. Furthermore, the L-type Ca2+ channel inhibitor, nifedipine, and the sarcoplasmic/endoplasmic reticulum Ca2+-ATPase (SERCA) pump inhibitor, thapsigargin, did not change the vasodilatory effect of linagliptin. SIGNIFICANCE: We suggests that linagliptin-induced vasodilation was mediated by the inhibition of Rho-associated kinase, but not with the endothelium, cAMP-PKA or cGMP-PKG-dependent signaling pathways, K+ channels, Ca2+ influx, or SERCA pump.

G2A Protects Mice against Sepsis by Modulating Kupffer Cell Activation: Cooperativity with Adenosine Receptor 2b.

G2A is a GPCR abundantly expressed in immune cells. G2A-/- mice showed higher lethality, higher plasma cytokines, and an impaired bacterial clearance in response to a murine model of sepsis (cecal ligation and puncture), which were blocked by GdCl3, an inhibitor of Kupffer cells. Anti-IL-10 Ab reversed the impaired bacterial clearance in G2A-/- mice. Indomethacin effectively blocked both the increased i.p. IL-10 levels and the impaired bacterial clearance, indicating that disturbed PG system is the proximal cause of these phenomena. Stimulation with LPS/C5a induced an increase in Escherichia coli phagocytosis and intracellular cAMP levels in G2A+/+ peritoneal macrophages but not G2A-/- cells, which showed more PGE2/nitrite release and intracellular reactive oxygen species levels. Heterologous coexpression of G2A and adenosine receptor type 2b (A2bAR) induced a synergistic increase in cAMP signaling in a ligand-independent manner, with the evidence of physical interaction of G2A with A2bAR. BAY 60-6583, a specific agonist for A2bAR, increased intracellular cAMP levels in Kupffer cells from G2A+/+ but not from G2A-/- mice. Both G2A and A2bAR were required for antiseptic action of lysophosphatidylcholine. These results show inappropriate activation of G2A-/- Kupffer cells to septic insults due to an impaired cAMP signaling possibly by lack of interaction with A2bAR.