The ethyl acetate fractions prepared from the leaves of Thuja orientalis significantly inhibited nitric oxide (NO) production in lipopolysaccharide-stimulated BV2 microglial cells. According to bioassay-coupled LC-QTOF MS/MS, the components near 22 and 25 mins in the mass chromatogram highly inhibited NO production and were expected to be labdane diterpenes, and the active components were characterized via further isolation. The results of the NO production inhibitory assay of the isolated compounds correlated well with the results of bioassay-coupled LC-QTOF MS/MS. Among the identified constituents, NO production inhibitory activities of 16-hydroxy-labda-8(17),13-diene-15,19-dioic acid butenolide (2) and 15-hydroxypinusolidic acid (3) were newly reported. Taken together, these results demonstrated that LC-QTOF MS/MS coupled with NO production inhibition assay was a powerful tool for accurately predicting new anti-inflammatory constituents in the extracts from natural products. Moreover, it provided a potential basis for broadening the application of bioassay-coupled LC-QTOF MS/MS in natural product research.

The proliferation, migration, cytokine release, and contraction of airway smooth muscle cells are key events in the airway remodeling process that occur in lung disease such as asthma, chronic obstruction pulmonary disease, and cancer. These events can be modulated by a number of factors, including cigarette smoke extract (CSE). CSE-induced alterations in the viability, migration, and contractile abilities of normal human airway cells remain unclear. This study investigated the effect of CSE on cell viability, migration, tumor necrosis factor (TNF)-alpha secretion, and contraction in normal human bronchial smooth muscle cells (HBSMCs). Treatment of HBSMCs with 10% CSE induced cell death, and the death was accompanied by the generation of reactive oxygen species (ROS). CSE-induced cell death was reduced by N-acetyl-l-cysteine (NAC), an ROS scavenger. In addition, CSE reduced the migration ability of HBSMCs by 75%. The combination of NAC with CSE blocked the CSE-induced reduction of cell migration. However, CSE had no effect on TNF-alpha secretion and NF-kappaB activation. CSE induced an increase in intracellular Ca2+ concentration in 64% of HBSMCs. CSE reduced the contractile ability of HBSMCs, and the ability was enhanced by NAC treatment. These results demonstrate that CSE treatment induces cell death and reduces migration and contraction by increasing ROS generation in normal HBSMCs. These results suggest that CSE may induce airway change through cell death and reduction in migration and contraction of normal HBSMCs.
Acetylcysteine ; Airway Remodeling ; Asthma ; Bronchioles ; Cell Death ; Cell Movement ; Cell Survival ; Contracts ; Emigration and Immigration ; Humans ; Lung Diseases ; Muscle, Smooth ; Myocytes, Smooth Muscle ; NF-kappa B ; Reactive Oxygen Species ; Smoke ; Tobacco Products ; Tumor Necrosis Factor-alpha

Background: Suicide is one of the leading causes of death in the military as in the general population. To mitigate such a serious public health problem, identifying the risk or protective factors of suicide behaviors is crucial. Methods: We analyzed the representative data of the 2014 Korean Armed Forces to explore the relationship between past year adverse events (PAE), accumulated lifetime trauma (ALT), mental illness vulnerability, perceived social support, and suicidal ideation in the previous year. Results: Among the 6,377 subjects, 3.7% of males and 6.2% of females reported suicidal ideation in the previous year. Multivariate analytic models identified significant associations of PAE and ALT with suicidal ideation with a dose-response pattern. The mental illness vulnerability showed the most significant association with suicidal ideation even after controlling PAE or ALT. We found that perceived social support may be potentially linked with a reduced risk of suicidal ideation. Conclusion This Korean military representative data demonstrates mental illness vulnerability; PAE; and lifetime trauma as significant risk factors of suicidal ideation, while perceived social support was found as a potential protective factor. Given the importance of the prevention of suicide in the military, those risk and protective factors may be used to screen soldiers at risk of suicide and provide further support on mental health services as needed.

Background: Suicide is one of the leading causes of death in the military as in the general population. To mitigate such a serious public health problem, identifying the risk or protective factors of suicide behaviors is crucial. Methods: We analyzed the representative data of the 2014 Korean Armed Forces to explore the relationship between past year adverse events (PAE), accumulated lifetime trauma (ALT), mental illness vulnerability, perceived social support, and suicidal ideation in the previous year. Results: Among the 6,377 subjects, 3.7% of males and 6.2% of females reported suicidal ideation in the previous year. Multivariate analytic models identified significant associations of PAE and ALT with suicidal ideation with a dose-response pattern. The mental illness vulnerability showed the most significant association with suicidal ideation even after controlling PAE or ALT. We found that perceived social support may be potentially linked with a reduced risk of suicidal ideation. Conclusion This Korean military representative data demonstrates mental illness vulnerability; PAE; and lifetime trauma as significant risk factors of suicidal ideation, while perceived social support was found as a potential protective factor. Given the importance of the prevention of suicide in the military, those risk and protective factors may be used to screen soldiers at risk of suicide and provide further support on mental health services as needed.

BACKGROUND AND OBJECTIVES: Recent studies have shown that many kinds of K+ channels, including the muscarinic K+ channel (KACh), are activated in the ischemic heart. It is known that these channels can be modulated by phosphorylation. However, little is known about the function of the KACh in ischemic hearts. In this study, we examined whether the KACh channel is mediated by protein kinase C (PKC) activation in rat atrial myocytes under ischemic conditions. MATERIALS AND METHODS: Single atrial cells of adult rat heart were prepared by collagenase digestion. Channel activity of KACh was recorded by cell-attached configuration from single atrial cells under ischemic conditions, using a patch clamp technique. To simulate ischemia, adenosine or potassium cyanide (KCN) was applied to atrial myocytes, and Western blot was performed to specify PKC isoforms. RESULTS: Adenosine and KCN markedly increased KACh channel activity. The responses to adenosine and KCN were increased 3-fold at mean open time from that observed with control. Channel activity of KACh was blocked by pretreatment with PKC antagonists such as sphingosine, Go 6976, and rottlerin. PKC alpha and PKC betaI isoform levels were increased in the membrane fraction of ischemic heart, indicating that ischemic stress might trigger translocation of cytosolic PKC to the cell membrane. CONCLUSION: These results show that KACh channels are modulated by PKC activation under ischemic conditions induced by adenosine or KCN. Therefore, the channels can protect the heart from ischemic stress by increasing channel activity.
Adenosine ; Adult ; Animals ; Blotting, Western ; Cell Membrane ; Collagenases ; Cytosol ; Digestion ; Heart ; Humans ; Ischemia ; Membranes ; Muscle Cells* ; Phosphorylation ; Potassium Cyanide ; Protein Isoforms ; Protein Kinase C* ; Protein Kinases* ; Rats* ; Sphingosine

Endometritis is one of the primary reasons for reproductive failure. In order to investigate endometritis-associated marker proteins, proteomic analysis was performed on bovine endometrium with endometritis. In bovine endometritis, desmin, alpha-actin-2, heat-shock protein (HSP) 27, peroxiredoxin-6, luteinizing hormone receptor isoform 1, collectin-43 precursor, deoxyribonuclease-I (DNase-I), and MHC class I heavy chain (MHC-Ih) were up-regulated. In contrast, transferrin, interleukin-2 precursor, hemoglobin beta subunit, and potassium channel tetramerisation domain-containing 11 (KCTD11) were down-regulated in comparison to normal endometrium. The proteomic results were validated by semiquantitative-PCR and immunoblot analysis. The mRNA levels of desmin, transferrin, alpha-actin-2, HSP27, KCTD11, and MHC-Ih were up-regulated by over 1.5-fold, and showed a pattern similar to their proteomic profiles. Desmin and alpha-actin-2 protein showed positive correlations between proteomic analysis and immunoblot analysis. These results suggest that desmin and alpha-actin-2 may play important roles in endometritis-related function, and could be useful markers for the diagnosis of bovine endometritis.
Actins ; Collectins ; Desmin ; Endometritis ; Endometrium ; Female ; Heat-Shock Proteins ; Hemoglobins ; Interleukin-2 ; Potassium Channels ; Proteins ; Proteomics ; Receptors, LH ; RNA, Messenger ; Transferrin

Single-channel recordings of TASK-1 and TASK-3, members of two-pore domain K+ channel family, have not yet been reported in dorsal root ganglion (DRG) neurons, even though their mRNA and activity in whole-cell currents have been detected in these neurons. Here, we report single-channel kinetics of the TASK-3-like K+ channel in DRG neurons and up-regulation of TASK-3 mRNA expression in tissues isolated from animals with spinal cord injury (SCI). In DRG neurons, the single-channel conductance of TASK-3-like K+ channel was 33.0+/-0.1 pS at -60 mV, and TASK-3 activity fell by 65+/-5% when the extracellular pH was changed from 7.3 to 6.3, indicating that the DRG K+ channel is similar to cloned TASK-3 channel. TASK-3 mRNA and protein levels in brain, spinal cord, and DRG were significantly higher in injured animals than in sham-operated ones. These results indicate that TASK-3 channels are expressed and functional in DRG neurons and the expression level is up-regulated following SCI, and suggest that TASK-3 channel could act as a potential background K+ channel under SCI-induced acidic condition.
Acidosis ; Animals ; Brain ; Clone Cells ; Diagnosis-Related Groups ; Ganglia, Spinal ; Humans ; Hydrogen-Ion Concentration ; Kinetics ; Neurons ; Rats ; RNA, Messenger ; Spinal Cord ; Spinal Cord Injuries ; Spinal Nerve Roots ; Up-Regulation

OBJECTIVE: This study investigated whether the resilience of males with probable bipolar depression (PBD) can be strengthened and compared it to that of males with probable unipolar depression (PUD). METHODS: Prospective data for 198 participants (PBD: 66, PUD: 66, normal control: 66) were analyzed. The participants' resilience, bipolarity and severity of depressive symptoms were evaluated at baseline and after 5 weeks. Analysis of variance (ANOVA) and repeated measure ANOVA was performed for comparing resilience between three groups through a basic military training. RESULTS: The PBD group demonstrated more resilience than the PUD group at baseline. Participants with PBD became significantly more resilient than participants with PUD after 5 weeks (p<0.01, F=6.967, η²(p)=0.052). CONCLUSION: The study indicates that interventions that strengthen resilience need to be developed for males with PBD and that such interventions are more effective for males with PBD than PUD.
Bipolar Disorder* ; Depression ; Depressive Disorder* ; Humans ; Male* ; Military Personnel ; Prospective Studies

Background: The urinary levels of volatile organic compound (VOC) metabolites provide individual exposure levels compared to data obtained by measuring these compounds in ambient air. We aimed to investigate the association between personal urinary concentrations of VOC metabolites and symptoms of atopic dermatitis in schoolchildren. Methods: Nine urinary VOC metabolites were analyzed from urine samples of 149 children. Diagnosis of atopic dermatitis was determined using standardized questionnaires. Pediatricians visited the schools and rated the severity of symptoms using the SCORing Atopic Dermatitis (SCORAD) in all children. Results: Forty-five children (30.2%) had atopic dermatitis based on the International Study of Asthma and Allergies in Childhood (ISAAC) results and 35 children (23.8%) had symptoms of atopic dermatitis with positive SCORAD index values (defined as SCORAD ≥ 5). Children with benzylmercapturic acid detected in personal urines were associated with presence of atopic dermatitis and positive SCORAD index values. Children in the highest quartile of mandelic acid concentration were associated with presence of atopic dermatitis and positive SCORAD results. Conclusion Personal exposure to VOCs, as indicated by urinary levels of VOC metabolites, was associated with presence of atopic dermatitis and the SCORAD index value.

Neuropathic pain is a complex state showing increased pain response with dysfunctional inhibitory neurotransmission. The TREK family, one of the two pore domain K+ (K2P) channel subgroups were focused among various mechanisms of neuropathic pain. These channels influence neuronal excitability and are thought to be related in mechano/thermosensation. However, only a little is known about the expression and role of TREK-1 and TREK-2, in neuropathic pain. It is performed to know whether TREK-1 and/or 2 are positively related in dorsal root ganglion (DRG) of a mouse neuropathic pain model, the chronic constriction injury (CCI) model. Following this purpose, Reverse Transcription Polymerase Chain Reaction (RT-PCR) and western blot analyses were performed using mouse DRG of CCI model and compared to the sham surgery group. Immunofluorescence staining of isolectin-B4 (IB4) and TREK were performed. Electrophysiological recordings of single channel currents were analyzed to obtain the information about the channel. Interactions with known TREK activators were tested to confirm the expression. While both TREK-1 and TREK-2 mRNA were significantly overexpressed in DRG of CCI mice, only TREK-1 showed significant increase (~9 fold) in western blot analysis. The TREK-1-like channel recorded in DRG neurons of the CCI mouse showed similar current-voltage relationship and conductance to TREK-1. It was easily activated by low pH solution (pH 6.3), negative pressure, and riluzole. Immunofluorescence images showed the expression of TREK-1 was stronger compared to TREK-2 on IB4 positive neurons. These results suggest that modulation of the TREK-1 channel may have beneficial analgesic effects in neuropathic pain patients.
Animals ; Blotting, Western ; Constriction* ; Diagnosis-Related Groups ; Fluorescent Antibody Technique ; Ganglia, Spinal* ; Humans ; Hydrogen-Ion Concentration ; Mice* ; Neuralgia* ; Neurons ; Polymerase Chain Reaction ; Reverse Transcription ; Riluzole ; RNA, Messenger ; Spinal Nerve Roots* ; Synaptic Transmission