Functional Characterization of Gonadotropin-Releasing Hormone and Corazonin Signaling Systems in Pacific Abalone: Toward Reclassification of Invertebrate Neuropeptides.

INTRODUCTION: The proposed evolutionary origins and corresponding nomenclature of bilaterian gonadotropin-releasing hormone (GnRH)-related neuropeptides have changed tremendously with the aid of receptor deorphanization. However, the reclassification of the GnRH and corazonin (CRZ) signaling systems in Lophotrochozoa remains unclear. METHODS: We characterized GnRH and CRZ receptors in the mollusk Pacific abalone, Haliotis discus hannai (Hdh), by phylogenetic and gene expression analyses, bioluminescence-based reporter, Western blotting, substitution of peptide amino acids, in vivo neuropeptide injection, and RNA interference assays. RESULTS: Two Hdh CRZ-like receptors (Hdh-CRZR-A and Hdh-CRZR-B) and three Hdh GnRH-like receptors (Hdh-GnRHR1-A, Hdh-GnRHR1-B, and Hdh-GnRHR2) were identified. In phylogenetic analysis, Hdh-CRZR-A and -B grouped within the CRZ-type receptors, whereas Hdh-GnRHR1-A/-B and Hdh-GnRHR2 clustered within the GnRH/adipokinetic hormone (AKH)/CRZ-related peptide-type receptors. Hdh-CRZR-A/-B and Hdh-GnRHR1-A were activated by Hdh-CRZ (pQNYHFSNGWHA-NH2) and Hdh-GnRH (pQISFSPNWGT-NH2), respectively. Hdh-CRZR-A/-B dually coupled with the Gαq and Gαs signaling pathways, whereas Hdh-GnRHR1-A was linked only with Gαq signaling. Analysis of substituted peptides, [I2S3]Hdh-CRZ and [N2Y3H4]Hdh-GnRH, and in silico docking models revealed that the N-terminal amino acids of the peptides are critical for the selectivity of Hdh-CRZR and Hdh-GnRHR. Two precursor transcripts for Hdh-CRZ and Hdh-GnRH peptides and their receptors were mainly expressed in the neural ganglia, and their levels increased in starved abalones. Injection of Hdh-CRZ peptide into abalones decreased food consumption, whereas Hdh-CRZR knockdown increased food consumption. Moreover, Hdh-CRZ induced germinal vesicle breakdown in mature oocytes. CONCLUSION: Characterization of Hdh-CRZRs and Hdh-GnRHRs and their cognate peptides provides new insight into the evolutionary route of GnRH-related signaling systems in bilaterians.

Associations of personal urinary volatile organic compounds and lung function in children.

BACKGROUND: We investigated the correlation between urine VOC metabolites and airway function in children exposed to anthropogenic volatile organic compounds (VOCs), notable pollutants impacting respiratory health. METHODS: Out of 157 respondents, 141 completed skin prick tests, spirometry, IOS, and provided urine samples following the International Study of Asthma and Allergies in Childhood (ISAAC)-related questions. Allergic sensitization was assessed through skin prick tests, and airway functions were evaluated using spirometry and impulse oscillometry (IOS). Forced expiratory volume in 1 s (FEV1), forced vital capacity (FVC) was recorded and FEV1/FVC ratio was calculated. Airway mechanics parameters including respiratory resistance at 5 Hz (Rrs5) mean respiratory resistance between 5 Hz and 20 Hz (Rrs5-20), were also recorded. Urine concentrations of metabolites of benzene, ethylbenzene, toluene, xylene, styrene, formaldehyde, carbon-disulfide were analyzed by gas chromatography/tandem mass spectroscopy. RESULTS: The median age at study participation was 7.1 (SD 0.3) years. Muconic acid (benzene metabolites) and o-methyl-hippuric acid (xylene metabolites) above medians were associated with a significant increase in Rrs5 (muconic acid: aß = 0.150, p = .002; o-methyl-hippuric acid: aß = 0.143, p = .023) and a decrease in FEV1/FVC (o-methyl-hippuric acid: aß = 0.054, p = .028) compared to those below median. No associations were observed for Rrs5-20 and FEV1 between the groups categorized as above and below the median (all parameter p values > .05). CONCLUSIONS: Elevated levels of benzene and xylene metabolites were associated with a significant increase in Rrs5 and a decrease in FEV1/FVC, related to increased resistance and restrictive lung conditions compared to individuals with concentrations below the median.

Characterization of myoinhibitory peptide signaling system and its implication in larval metamorphosis and spawning behavior in Pacific abalone.

Myoinhibitory peptides (MIPs) affect various physiological functions, including juvenile hormone signaling, muscle contraction, larval development, and reproduction in invertebrates. Although MIPs are ligands for MIP and/or sex peptide receptors (MIP/SPRs) in diverse arthropods and model organisms belonging to Lophotrochozoa, the MIP signaling system has not yet been fully investigated in mollusks. In this study, we identified the MIP signaling system in the Pacific abalone Haliotis discus hannai (Hdh). Similar to the invertebrate MIPs, a total of eight paracopies of MIPs (named Hdh-MIP1 to Hdh-MIP8), harboring a WX5-7Wamide motif, except for Hdh-MIP2, were found in the Hdh-MIP precursor. Furthermore, we characterized a functional Hdh-MIPR, which responded to the Hdh-MIPs, except for Hdh-MIP2, possibly linked with the PKC/Ca2+ and PKA/cAMP signaling pathways. Hdh-MIPs delayed larval metamorphosis but increased the spawning behavior. These results suggest that the Hdh-MIP signaling system provides insights into the unique function of MIP in invertebrates.

Crustacean cardioactive peptide signaling system in the gastropod mollusk Pacific abalone.

Crustacean cardioactive peptide (CCAP) signaling systems have been characterized in a diverse range of protostomes, representatively in arthropods. The cyclic CX5C-type CCAP regulates various biological activities through CCAP receptors (CCAPRs), which are orthologous to neuropeptide S receptors (NPSRs) in deuterostomes. However, the CCAPRs of the lophotrochozoa remain poorly characterized; therefore, the relationship between the CCAP, NPS, and CX4C-type oxytocin/vasopressin (OT/VP) signaling systems is unclear. In this study, we identified a CCAP precursor and two CCAPR isoforms in the Pacific abalone (Haliotis discus hannai; Hdh). The Hdh-CCAP precursor was found to harbor three CX5C-type and one CX4C-type CCAPs. The Hdh-CCAPRs displayed homology with protostome CCAPRs and deuterostome NPSRs, having characteristics of the rhodopsin-type G protein-coupled receptors. Phylogenetic analysis showed that lophotrochozoan CCAPRs, including Hdh-CCAPRs, form a monophyletic group distinct from arthropod CCAPRs. Reporter assays demonstrated that all examined Hdh-CCAPs and insect CCAP-induced intracellular Ca2+ mobilization and cAMP accumulation in Hdh-CCAPR-expressing HEK293 cells, whereas none of the CCAP peptides inhibited the forskolin-stimulated cAMP signaling pathway even at micromolar concentrations. In silico ligand-receptor docking models showed that the N-terminal FCN motifs of Hdh-CCAPs are deeply inserted inside the binding pocket of Hdh-CCAPR, forming extensive hydrophobic interactions. In mature Pacific abalone, the transcripts for Hdh-CCAP precursor and Hdh-CCAPR were highly expressed in the neural ganglia compared to the peripheral tissues. Collectively, this study characterized the first CCAP signaling system linked to both Ca2+/PKC and cAMP/PKA signal transduction pathways in gastropod mollusks and gives insights into the evolutional origins of deuterostomian NPS and OT/VP signaling systems.

Pneumonia Prevalence Upon Chest Radiography According to Vaccination Status Among Patients Under 50 Years of Age With Coronavirus Disease 2019.

BACKGROUND: Coronavirus disease 2019 (COVID-19) vaccination is effective in preventing the disease transmission and progression. However, the relatively mild disease course of the omicron variant and the decrease in antibodies over time after vaccination raise questions about the effectiveness of vaccination, especially in young people. We compared the prevalence of pneumonia and chest X-ray severity score according to vaccination status among patients < 50 years old with COVID-19. METHODS: From January 17 to March 17, 2022, 579 patients with COVID-19, who were < 50 years old and had a known vaccination history in our institution, were all included in this study. All patients underwent initial chest radiography, and follow-up chest radiographs were obtained every two days until discharge. Pneumonia was scored from the radiographs using the Brixia scoring system. The scores of the six lung zones were added for a total score ranging from 0 to 18. Patients were divided into four groups according to 10-year age intervals. Differences between groups were analyzed using the χ² or Fisher's exact tests for categorical variables and the Kruskal-Wallis test or analysis of variance for continuous variables. RESULTS: Among patients aged 12-19 years, the prevalence of pneumonia did not differ depending on vaccination status (non-vaccinated vs. vaccinated, 1/47 [2.1%] vs. 1/18 [5.6%]; P = 0.577). Among patients in their 20s, the prevalence of pneumonia was significantly higher among non-vaccinated patients than among vaccinated patients (8/28, 28.6% vs. 7/138, 5.1%, P < 0.001), similar to patients in their 40s (32/52 [61.5%] vs. 18/138 [13.0%]; P < 0.001). The chest X-ray severity score was also significantly higher in non-vaccinated patients than that in vaccinated patients in their 20s to their 40s (P < 0.001), but not among patients aged 12-19 years (P = 0.678). CONCLUSION: In patients aged 20-49 years, vaccinated patients had a significantly lower prevalence of pneumonia and chest X-ray severity score than non-vaccinated patients.

Stratification of Nontuberculous Mycobacterial Disease Risk in Type 2 Diabetes Based on Metformin Use: a Population-Based Cohort Study in South Korea.

In a 1:4 case-control matched analysis of data from a nationwide population-based cohort in South Korea, we evaluated whether metformin use mitigates the risk of nontuberculous mycobacterial disease in patients with type 2 diabetes. Multivariable analysis revealed no significant association of metformin use with a diminished risk for incident nontuberculous mycobacterial disease in patients with type 2 diabetes.

Neutrophil-Lymphocyte Ratio and Monocyte-Lymphocyte Ratio According to the Radiologic Severity of Mycobacterium avium Complex Pulmonary Disease.

BACKGROUND: To date, no study has investigated whether the neutrophil-lymphocyte ratio (NLR) and monocyte-lymphocyte ratio (MLR) have a clinical value in Mycobacterium avium complex (MAC)-pulmonary disease (PD). METHODS: We aimed to assess whether the baseline NLR and MLR were different according to the severity of MAC-PD based on the radiologic classification by retrospectively analyzing 549 patients treated in a tertiary referral center in South Korea. RESULTS: Both NLR and MLR were significantly higher as 3.33 and 0.43 respectively in the fibrocavitary type, followed by 2.34 and 0.27 in the cavitary nodular bronchiectatic type and significantly lower as 1.88 and 0.23 in the non-cavitary nodular bronchiectatic type. CONCLUSION: The baseline NLR and MLR showed a distinct difference in accordance with the radiologic severity of MAC-PD.

Incidence of cancer after asthma development: 2 independent population-based cohort studies.

BACKGROUND: Evidence regarding the risk of cancer development after asthma diagnosis is controversial and inconclusive. OBJECTIVE: This study sought to determine whether asthma is associated with an increased risk for incident cancer. METHODS: Two independent, population-based, longitudinal cohorts were examined, and estimated hazard ratios were determined using Cox regression. One group consisted of an unmatched cohort of 475,197 participants and a propensity score-matched cohort of 75,307 participants from the National Health Insurance Service-National Sample Cohort (NHIS-NSC; claims-based data from 2003 to 2015). The other group consisted of 5,440 participants from the Ansan-Ansung cohort (interview-based data from 2001 to 2014). RESULTS: The NHIS-NSC matched cohort had 572,740 person-years of follow-up, 6,885 people with new asthma diagnoses, and 68,422 people without asthma diagnoses. Adults with asthma had a 75% greater risk of incident cancer overall. The excess risk for incident cancer was greatest during the first 2 years after asthma diagnosis, and this risk remained elevated throughout follow-up. Patients with nonatopic asthma had a greater risk of overall cancer than those with atopic asthma. A high cumulative dose of inhaled corticosteroids among asthma patients was associated with a 56% reduced risk of lung cancer, but had no effect on the risk of overall cancer. The results from the NHIS-NSC unmatched cohort and the Ansan-Ansung cohort were similar to the primary results from the NHIS-NSC matched cohort. CONCLUSIONS: Asthma development was associated with an increased risk of subsequent cancer in 2 different Korean cohorts. Our findings provide an improved understanding of the pathogenesis of asthma and its relationship with carcinogenesis and suggest that clinicians should be aware of the higher risk of incident cancer among patients with asthma.

Effects of Radix Polygalae on Cognitive Decline and Depression in Estradiol Depletion Mouse Model of Menopause.

Postmenopausal syndrome refers to symptoms caused by the gradual decrease in female hormones after mid-40 years. As a target organ of estrogen, decrease in estrogen causes various changes in brain function such as a decrease in choline acetyltransferase and brain-derived neurotrophic factor; thus, postmenopausal women experience cognitive decline and more depressive symptoms than age-matched men. Radix Polygalae has been used for memory boosting and as a mood stabilizer and its components have shown neuroprotective, antidepressant, and stress relief properties. In a mouse model of estrogen depletion induced by 4-vinylcyclohexene diepoxide, Radix Polygalae was orally administered for 3 weeks. In these animals, cognitive and depression-related behaviors and molecular changes related to these behaviors were measured in the prefrontal cortex and hippocampus. Radix Polygalae improved working memory and contextual memory and despair-related behaviors in 4-vinylcyclohexene diepoxide-treated mice without increasing serum estradiol levels in this model. In relation to these behaviors, choline acetyltransferase and brain-derived neurotrophic factor in the prefrontal cortex and hippocampus and bcl-2-associated athanogene expression increased in the hippocampus. These results implicate the possible benefit of Radix Polygalae in use as a supplement of estrogen to prevent conditions such as postmenopausal depression and cognitive decline.

Association of phthalate exposure and airway dysfunction with mediation by serum periostin.

BACKGROUND: Phthalates can cause respiratory and immunological disorders. However, little is known about the role of serum periostin and YKL-40 levels in mediating the effects of phthalates. We investigated the mediating role of these biomarkers in the relationship between phthalates and airway dysfunction. METHODS: A total of 487 children (aged 10-12 years old) were examined. Four high-molecular-weight phthalate (HMWP) [Σ4 HMWP] metabolites and 3 low-molecular-weight phthalate (LMWP) [Σ3 LMWP] metabolites in urine samples were measured. Serum periostin and YKL-40 levels were measured. Airway function was measured using impulse oscillometry. A mediation model was used to quantify the mediating effects of periostin and YKL-40 on airway dysfunction. RESULTS: After adjustment for height, gender, BMI z-score, aeroallergen sensitization, secondary smoking, and vitamin D level, the level of urinary Σ3 LMWP metabolites was significantly associated with respiratory system resistance at 5 Hz (Rrs5; adjusted ß: 0.020, 95% CI: 0.005-0.034; p = .010). The levels of urinary Σ4 HMWP and Σ3 LMWP metabolites were significantly associated with periostin level, but not with YKL-40 level. In addition, the periostin level was associated with Rrs5 (adjusted ß: 0.048, 95% CI: 0.015-0.081; p = .005) and Rrs20-5 (adjusted ß: 0.040, 95% CI: 0.011-0.069; p = .007). Serum periostin level had a significant effect in mediating the relationship between Σ3 LMWP and Rrs5 (13.9%, 95% CI: 10.7-77.0; p < .001). CONCLUSION: Exposure to LMWPs was significantly associated with airway dysfunction, and this effect was partially attributable to increased serum periostin level.

Protaetiibacter larvae sp. nov. and Agromyces intestinalis sp. nov., isolated from the gut of larvae of Protaetia brevitarsis seulensis, reclassification of Lysinimonas yzui as Pseudolysinimonas yzui comb. nov. and emended description of the genus Pseudolysinimonas.

Two bacterial strains, FWR-8T and CFWR-9T, were isolated from the gut of larvae of Protaetia brevitarsis seulensis that were raised at the National Institute of Agricultural Sciences, Wanju-gun, Republic of Korea. Both strains were strictly aerobic, Gram-stain-positive and non-motile. Strain FWR-8T possessed the highest sequence similarity (98.7 %) to that of Protaetiibacter intestinalis 2DFWR-13T and the phylogenetic tree revealed that strain FWR-8T formed a cluster with Ptb. intestinalis 2DFWR-13T. Pseudolysinimonas kribbensis MSL-13T and Lysinimonas yzui N7XX-4T shared a high 16S rRNA gene sequence similarity (97.8 %) and formed a cluster adjacent to the cluster that included Ptb. intestinalis 2DFWR-13T. The 16S rRNA gene sequence of strain CFWR-9T exhibited the highest similarity (97.7 %) to that of Agromyces binzhouensis OAct353T and the phylogenetic tree indicated that strain CFWR-9T formed one independent cluster with A. binzhouensis OAct353T that was within the radius of the genus Agromyces. The peptidoglycan type, major fatty acids, major menaquinones and total polar lipids of strain FWR-8T were characterized as type B1, iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0, MK-15, MK-16 and MK-14, and diphosphatidylglycerol, phosphatidylglycerol, two unidentified glycolipids and one unidentified lipid, respectively. Those from strain CFWR-9T were type B1, iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0, MK-11, MK-12 and MK-10, and diphosphatidylglycerol, phosphatidylglycerol, two unidentified glycolipids and one unidentified lipid, respectively. Based on the phenotypic and genotypic data, strains FWR-8T and CFWR-9T each represent a novel species within the genera Protaetiibacter and Agromyces, respectively. For these species, the names Protaetiibacter larvae sp. nov. and Agromyces intestinalis sp. nov. have been proposed, with the type strains FWR-8T (=KACC 19322T=NBRC 113051T) and CFWR-9T (=KACC 19306T=NBRC 113046T), respectively. Our results also justify a reclassification of Lysinimonas yzui as Pseudolysinimonas yzui comb. nov. and an emended description of the genus Pseudolysinimonas isprovided.

Microbacterium protaetiae sp. nov., isolated from gut of larva of Protaetia brevitarsis seulensis.

A Gram-stain-positive, strictly aerobic, polar flagellated, short rod-shaped bacterium, designated DFW100M-13T, was isolated from gut of the larva of Protaetia brevitarsis seulensis collected from Wanju-gun, South Korea. The growth range of NaCl concentration was 0-3 % (w/v) (optimally 0 % (w/v)), the temperature range for growth was 10-40 °C (optimally 28-30 °C), and the pH range for growth was pH 6.0-9.0 (optimally pH 7.0-8.0). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain DFW100M-13T had a high sequence similarity to members of the genus Microbacterium, having the highest similarity with Microbacterium luticocti DSM 19459T (97.7 %), Microbacterium rhizosphaerae CHO1T (97.1 %), and Microbacterium immunditiarum SK 18T (97.0 %), and formed a distinct lineage with Microbacterium luticocti DSM 19459T within the genus Microbacterium. A phylogenetic tree based on house-keeping genes also showed the result similar to the 16S rRNA gene-based tree. The main respiratory quinone (>10 %) was MK-11, MK-12 and MK-10, and the predominant cellular fatty acids (>10 %) were iso-C16 : 0, anteiso-C17 : 0 and anteiso-C15 : 0. The polar lipids were composed of diphosphatidylglycerol, phosphatidylglycerol, an inidentified glycolipid and an unidnetified lipid. The peptidoglycan type was supposed to be the B2ß with amino acids d-alanine, d-glutamic acid, glycine, l-homoserine and d-ornithine. The genomic DNA G+C content was 68.0 mol%. Based on the polyphasic taxonomic data, strain DFW100M-13T is considered to represent a novel species, for which the name Microbacterium protaetiae sp. nov. is proposed. The type strain is DFW100M-13T (=KACC 19323T=NBRC 113120T).

Lactococcus protaetiae sp. nov. and Xylanimonas protaetiae sp. nov., isolated from gut of larvae of Protaetia brevitarsis seulensis.

Two bacterial strains, designated 2DFWM-2T and FW10M-9T, were isolated from gut of larva of Protaetia brevitarsis seulensis grown at the National Institute of Agricultural Sciences, Wanju-gun, South Korea. 16S rRNA and rpoB gene sequences showed that strain 2DFWM-2T formed a separate branch with Lactococcus allomyrinae 1JSPR-7T in the genus Lactococcus, adjacent to a group of Lactococcus lactis subspecies. ANI and dDDH values between 2DFWM-2T and Lactococcus allomyrinae 1JSPR-7T were 93.30% and 53.20%, respectively. Based on the 16S rRNA gene sequence analysis, strain FW10M-9T was classified into the genus Xylanimonas revealing 96.9-98.5% sequence similarities with the Xylanimonas species. The ANI values of strain FW10M-9T with the closely species Xylanimonas pachnodae NBRC 107786T, Xylanimonas allomyrinae 2JSPR-7T, Isoptericola variabilis JCM 11754T and Xylanimonas cellulosilytica DSM 15894T was 81.5%, 81.2%, 81.0% and 84.1%, respectively, and the dDDH values estimated by GGDC was 24.3%, 24.3%, 29.3% and 28.1%, respectively. On the basis of the phenotypic and genotypic data, it is proposed that strain 2DFWM-2T represents a novel species of the genus Lactococcus, for which the name Lactococcus protaetiae sp. nov. is proposed, and the type strain is 2DFWM-2T (= KACC 19320T = NBRC 113069T). And, strain FW10M-9T represents a novel species of the genus Xylanimonas, for which the name Xylanimonas protaetiae sp. nov. is proposed, and the type strain is FW10M-9T (= KACC 19331T = NBRC 113053T).

Impact of Body Mass Index Change on the Prognosis of Chronic Obstructive Pulmonary Disease.

BACKGROUND: Low body mass index (BMI) is an important prognostic factor in chronic obstructive pulmonary disease (COPD). However, the prognostic value of longitudinal BMI change in COPD has not been well studied. OBJECTIVE: We aimed to evaluate the association between longitudinal change of BMI and prognosis of COPD in Korean COPD cohort. METHODS: This study was conducted in a prospective Korean Obstructive Lung Disease (KOLD) cohort where COPD patients were recruited on an outpatient basis at 17 hospitals in South Korea. Annual BMI was measured over a period of 3 years or more. All patients were categorized into underweight (UW), normal weight (NW), and overweight (OW) groups by BMI. Clinical characteristics and outcomes including exacerbation and mortality were compared based on initial BMI grade and longitudinal change of BMI. RESULTS: This analysis included 537 COPD patients (mean age = 67.4 ± 7.9 years, male = 97.0%, mean BMI = 23.0 ± 3.1) of KOLD cohort. The proportions of UW, NW, and OW groups were 6.9% (n = 37), 68.9% (n = 370), and 24.2% (n = 130) respectively. The UW group showed lower forced expiratory volume in 1 s (FEV1) (p < 0.001), shorter 6-minute walk distance (p < 0.001), higher modified Medical Research Council score (p = 0.002), higher St. George Respiratory Questionnaire score (p < 0.001), higher emphysema index (p < 0.001) and air-trapping index (p < 0.001), and more frequent (p < 0.001) and severe exacerbations (p = 0.003). Multivariable analyses demonstrated that decrease of BMI (hazard ratio [HR] = 0.786, p = 0.038) and the descent of BMI group (HR = 3.167, p = 0.016) at 3-year follow-up along with age, initial BMI, post-bronchodilator FEV1, and severe exacerbations were significantly associated with mortality. CONCLUSIONS: This study demonstrated that BMI decrease during follow-up was independently associated with exacerbation and higher mortality of COPD, suggesting BMI reduction in COPD should be carefully managed.

Protective Effect of Tetrahydroquinolines from the Edible Insect Allomyrina dichotoma on LPS-Induced Vascular Inflammatory Responses.

The larva of Allomyrina dichotoma (family Scarabaeidae) is an edible insect that is registered in the Korean Food Standards Codex as a food resource. The chemical study on the larvae of A. dichotoma resulted in the isolation of three new tetrahydroquinolines, allomyrinaines A-C (1-3), one new dopamine derivative, allomyrinamide A (4), and four known compounds (5-8). The structures were elucidated on the basis of 1D and 2D nuclear magnetic resonance (NMR) and MS spectroscopic data analysis. Allomyrinaines A-C (1-3) possessed three stereogenic centers at C-2, C-3, and C-4, whose relative configurations were determined by analyses of the coupling constants and the nuclear Overhauser enhancement spectroscopy (NOESY) data, as well as DP4+ calculation. The anti-inflammatory effects of compounds 1-4 were evaluated in human endothelial cells. Allomyrinaines A-C (1-3) could stabilize vascular barrier integrity on lipopolysaccharide (LPS)-induced vascular inflammation via inhibition of the nuclear factor-κB (NF-κB) pathway. The physiologically relevant concentration was confirmed by Q-TOF-MS-based quantitative analysis on allomyrinaines A-C in crude extract. This study suggests that allomyrinaines A-C (1-3) are bioactive constituents of A. dichotoma to treat vascular inflammatory disorder.

Extract of Boehmeria nivea Suppresses Mast Cell-Mediated Allergic Inflammation by Inhibiting Mitogen-Activated Protein Kinase and Nuclear Factor-κB.

Mast cells are effector cells that initiate allergic inflammatory immune responses by inducing inflammatory mediators. Boehmeria nivea (Linn.) Gaudich is a natural herb in the nettle family Urticaceae that possesses numerous pharmacological properties. Despite the various pharmacological benefits of Boehmeria nivea, its effects on allergic inflammation have not yet been determined. Here, we investigated the effect of the ethanol extract of Boehmeria nivea (BNE) on degranulation rat basophilic leukemia (RBL)-2H3 mast cells stimulated with anti-dinitrophenyl (anti-DNP) and bovine serum albumin (BSA) during immunoglobulin E (IgE)-mediated allergic immune response. The results showed inhibition of the release of ß-hexosaminidase and histamine from the cells. BNE suppressed pro-inflammatory cytokines (Tumor necrosis factor (TNF)-α, Interleukin (IL)-1ß, and IL-6) and reduced T helper (Th)2 cytokine IL-4 expression and/or secretion correlated with the downregulation of p38, extracellular signal-regulated kinases (ERK) mitogen-activated protein kinase (MAPK), and nuclear factor-κB (NF-κB) signaling pathways in treated RBL-2H3 mast cells. In passive cutaneous anaphylaxis, treatment with BNE during IgE-mediated local allergic reaction triggered a reduction in mouse ear pigmentation and thickness. Taken together, these results indicated that BNE suppressed mast cell-mediated inflammation, suggesting that BNE might be a candidate for the treatment of various allergic disorders.

Protaetiibacter intestinalis gen. nov., of the family Microbacteriaceae, isolated from gut of Protaetia brevitarsis seulensis, reclassification of Lysinimonas kribbensis Jang et al. 2013 as Pseudolysinimonas kribbensis gen. nov., comb. nov. and emended description of the genus Lysinimonas Jang et al. 2013.

A Gram-stain-positive, strictly aerobic, rod-shaped, non-spore-forming, non-motile bacterium, designated strain 2DFWR-13T, was isolated from gut of the larva of Protaetia brevitarsis seulensis, in the Republic of Korea. Strain 2DFWR-13T showed high sequence similarities to Lysinimonas kribbensis MSL-13T (97.7 %), Homoserinibacter gongjuensis 5GH26-15T (97.2 %), Microbacterium deminutum KV-483T (97.1 %) and Herbiconiux ginsengi CGMCC 4.3491T (97.1 %). The predominant fatty acids (>10 % of the total fatty acids) were anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. The major menaquinones were MK-13 and MK-12. The peptidoglycan type was type B2 with the diagnostic amino acid d-DAB. The N-acyl type of the murein was glycolyl. The polar lipids consisted of diphosphatidylglycerol, an unidentified glycolipid and an unidentified lipid. The DNA G+C content was 71.5 mol%. Based on its phylogenetic distinctiveness and distinguishing phenotypic characteristics, we conclude that strain 2DFWR-13T represents a novel genus and species of the family Microbacteriaceae, for which the name Protaetiibacter intestinalis gen. nov., sp. nov. is proposed. The type strain of Protaetiibacter intestinalis is 2DFWR-13T (=KACC 19321T=NBRC 113050T). In addition, an emended description of the genus LysinimonasJang et al. 2013 and the reclassification of Lysinimonas kribbensisJang et al. 2013 as Pseudolysinimonas kribbensis gen. nov., comb. nov. are proposed.

Molecular characterization, expression analysis, and functional properties of multiple 5-hydroxytryptamine receptors in Pacific abalone (Haliotis discus hannai).

Neurotransmitters such as serotonin (5-hydroxytryptamine; 5-HT) in the central nervous system regulate diverse physiological functions, including reproduction, feeding, learning, and memory, in diverse animal phyla. 5-HT and the 5-HT1 subtype receptor play important roles in sexual maturation and in the initiation of gamete release in mollusks. However, little is known about the involvement of other 5-HT receptor subfamilies in the reproduction process. In the present study, we identified the cDNAs encoding eight subtypes of 5-HT receptors from the ganglia tissues of the Pacific abalone Haliotis discus hannai (Mollusca; Gastropoda; Haliotidae), and examined the gonadal expression of the transcripts of 5-HT receptors. A phylogenetic analysis indicated that the molluskan 5-HT receptors are largely classified into four major clades: 5-HT1/5/7, 5-HT2, 5-HT4, and 5-HT6. Among the H. discus hannai (Hdh) 5-HT1-7 transcripts, Hdh5-HT1B, 4A, 4B, and 6 were the major subtypes detected in the mature ovary. Estradiol-17ß injection into the pedal sinus induced the downregulation of 5-HT4B and upregulation of 5-HT6 transcripts in the ovary of mature abalone within 72 h. In HEK293 cells overexpressing Hdh5-HT1B, forskolin-stimulated cAMP response element luciferase (CRE-Luc) reporter activity was inhibited by 5-HT in a dose-dependent manner, whereas serum response element luciferase (SRE-Luc) activity was not affected. In Hdh5-HT4A-expressing HEK293 cells, forskolin-stimulated CRE-Luc and SRE-Luc reporter activities were both marginally increased by treatment with a high dose of 5-HT. Our results provide new insights into the roles of 5-HT through diverse G protein-coupled 5-HT receptors in the reproductive process of mollusks.

Surgical Outcomes after Pulmonary Resection for Non-Small Cell Lung Cancer with Localized Pleural Seeding First Detected during Surgery.

OBJECTIVES: Curative resection is not indicated for non-small cell lung cancer (NSCLC) with pleural seeding, which is classified as stage IV (M1a) disease. However, some patients with a presumably resectable main tumor are diagnosed with localized pleural seeding during surgery. METHODS: A retrospective analysis was performed of 3,975 patients who underwent surgery for NSCLC from 2000 to 2011. Among these cases, 78 (2.0%) patients had unexpected pleural seeding detected during surgery. Exploration with pleural biopsy was performed in 42 of these patients (exploration-only group) and pulmonary resection, including for the main tumor, was performed in 36 cases (resection group; sublobar resection in 12, lobectomy in 21, and pneumonectomy in 3 patients). Survival and cancer progression rates were estimated using the Kaplan-Meier method. Cox proportional hazard regression was used to evaluate prognostic factors associated with survival. RESULTS: Adenocarcinoma was the predominant histological type in both the exploration and resection groups (88.1 and 86.1%, respectively). Epidermal growth factor receptor expression was detected in 22 (52.4%) patients of the exploration group and 21 (58.3%) patients of the resection group. Baseline characteristics including age, sex, comorbidity, pulmonary function, and clinical T/N status were not significantly different between the two groups. There were no postoperative deaths in either group but postoperative complications occurred in two (4.8%) patients of the exploration group and three (8.3%) patients of the resection group. The overall 3- and 5-year survival rates in the exploration group were 41.1 and 15.2%, respectively, with a median survival time (MST) of 33 months, whereas they were 66.7 and 42.7%, respectively, in the resection group, with a 52-month MST (p = 0.012). Local and regional progression-free rates were significantly different (p < 0.001 and p = 0.029, respectively) between groups, whereas no difference was seen in the distant metastasis rates (p = 0.957). In multivariate survival analysis, surgical resection was the only significant prognostic factor (p = 0.01). CONCLUSIONS: Pulmonary resection including the main tumor, regardless of resection extent, may increase long-term survival for NSCLC patients with localized pleural seeding first detected during surgery, without a significant increase in hospital mortality or morbidity.

Expression profile and reproductive regulation of APGWamide in Pacific abalone (Haliotis discus hannai).

Neuropeptides in the central nervous system regulate reproductive activities in vertebrates. Ala-Pro-Gly-Trp-NH2 (APGWamide), a neuromediator expressed in the neural ganglia of mollusks, controls sexual maturation and reproduction. To clarify the role of APGWamide in sexual behavior regulation and gamete cell maturation in mollusks, we cloned the cDNA of APGWamide precursor (Hdh-APGWamide) and examined the spatiotemporal expression of the transcript in the Pacific abalone Haliotis discus hannai. The 222-amino acid sequence of the precursor deduced from the cDNA sequence showed typical features of gastropod APGWamide precursors. Phylogenetic analysis revealed that Hdh-APGWamide is classified with other gastropod APGWamide precursors, which form a separate branch from those of the bivalves. Hdh-APGWamide mRNA was highly expressed in the neural ganglia in both sexes. In females, the three ganglia (pleuro-pedal ganglion, PPG; branchial ganglion, and cerebral ganglion) showed similar expression in immature and mature animals, whereas in males, the level in the PPG only was higher at maturity (P < 0.05). In vivo injection of APGWamide or 5-hydroxytryptamine (10-3 M) increased the frequency of spawning and the number of released sperm cells by mature males (P < 0.05), while concentrations above 10-7 M enhanced germinal vesicle breakdown in fully developed cultured oocytes (P < 0.05). Thus, the phylogenetic branch of the APGWamide precursor gene in Haliotidae was separate from the other branches under the phylum Mollusca, and this gene exhibited ganglion-specific expression, indicating that it may induce final maturation and spawning in both sexes of Haliotis spp.