BACKGROUND: Beating cardiomyocyte regeneration therapies have revealed as alternative therapeutics for heart transplantation. Nonetheless, the importance of nitric oxide (NO) in cardiomyocyte regeneration has been widely suggested, little has been reported concerning endogenous NO during cardiomyocyte differentiation. METHODS: Here, we used P19CL6 cells and a Myocardiac infarction (MI) model to confirm NO-induced protein modification and its role in cardiac beating. Two tyrosine (Tyr) residues of b2-tubulin (Y106 and Y340) underwent nitrosylation (Tyr-NO) by endogenously generated NO during cardiomyocyte differentiation from pre-cardiomyocyte-like P19CL6 cells. RESULTS: Tyr-NO-b2-tubulin mediated the interaction with Stathmin, which promotes microtubule disassembly, and was prominently observed in spontaneously beating cell clusters and mouse embryonic heart (E11.5d). In myocardial infarction mice, Tyr-NO-b2-tubulin in transplanted cells was closely related with cardiac troponin-T expression with their functional recovery, reduced infarct size and thickened left ventricular wall. CONCLUSION This is the first discovery of a new target molecule of NO, b2-tubulin, that can promote normal cardiac beating and cardiomyocyte regeneration. Taken together, we suggest therapeutic potential of Tyr-NO-b2-tubulin, for ischemic cardiomyocyte, which can reduce unexpected side effect of stem cell transplantation, arrhythmogenesis.

Evidence suggests that the human respiratory tract, as with the gastrointestinal tract, has evolved to its current state in association with commensal microbes. However, little is known about how the airway microbiome affects the development of airway immune system. Here, we uncover a previously unidentified mode of interaction between host airway immunity and a unique strain (AIT01) of Staphylococcus epidermidis, a predominant species of the nasal microbiome. Intranasal administration of AIT01 increased the population of neutrophils and monocytes in mouse lungs. The recruitment of these immune cells resulted in the protection of the murine host against infection by Pseudomonas aeruginosa, a pathogenic bacterium.Interestingly, an AIT01-secreted protein identified as GAPDH, a well-known bacterial moonlighting protein, mediated this protective effect. Intranasal delivery of the purified GAPDH conferred significant resistance against other Gram-negative pathogens (Klebsiella pneumoniae and Acinetobacter baumannii) and influenza A virus. Our findings demonstrate the potential of a native nasal microbe and its secretory protein to enhance innate immune defense against airway infections. These results offer a promising preventive measure, particularly relevant in the context of global pandemics.

Background/Aims: Accurate diagnosis and the effects of allergen-specific immunotherapy for pollinosis are greatly dependent on the potency and stability of the extract. This study aimed to examine factors, such as temperature and storage buffer composition, that affect the stability of allergen extracts from pollens of allergenic importance in Korea. Methods: We prepared four pollen allergen extracts from ragweed, mugwort, Japanese hop, and sawtooth oak, which are the most important causes of seasonal rhinitis in Korea. Changes of protein and major allergen concentration were measured over 1 year by Bradford assay, two-site enzyme-linked immunosorbent assay, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis after reconstitution of the lyophilized allergen extract in various buffers and stored at room temperature (RT, 18°C to 26°C) or refrigerated (4°C). Results: More than 90% of the original protein concentration in all four extracts examined was detected over 1 year when 50% glycerol was added and refrigerated, whereas 57.9% to 94.5% remained in the extracts at RT. The addition of 50% glycerol to the storage buffer was found to prevent protein degradation at RT. Amb a 1, a major allergen of ragweed, was almost completely degraded in 9 weeks at RT when reconstituted in a buffer without 50% glycerol. However, 55.6% to 92.8% of Amb a 1 content was detected after 1 year of incubation at 4°C in all buffer conditions except 0.3% phenol. Conclusions Addition of 50% glycerol as well as refrigeration was found to be important in increasing the shelf-life of allergen extracts from pollens of allergenic importance.

BACKGROUND: We developed skin prick test (SPT) reagents for common inhalant allergens that reflected the real exposure in Korea. The study aim was to evaluate diagnostic usefulness and allergen potency of our inhalant SPT reagents in comparison with commercial products. METHODS: We produced eight common inhalant allergen SPT reagents using total extract (Prolagen): Dermatophagoides farinae, Dermatophagoides pteronyssinus, oak, ragweed, mugwort, Humulus japonicus pollens, as well as cat and dog allergens. We compared the newly developed reagents with three commercially available SPT reagents (Allergopharma, Hollister-Stier, Lofarma). We measured total protein concentrations, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), major allergen concentration, and biological allergen potencies measured by immunoglobulin E (IgE) immunoblotting and ImmunoCAP inhibition test. RESULTS: Diagnostic values of these SPT reagents were expressed as positivity rate and concordance rate of the results from ImmunoCAP allergen-specific IgE test in 94 allergic patients. In vitro analysis showed marked differences in protein concentrations, SDS-PAGE features, major allergen concentrations, and biological allergen potencies of four different SPT reagents. In vivo analysis showed that positive rates and concordance rates of Prolagen® SPT reagents were similar compared to the three commercial SPT reagents. CONCLUSION: The newly developed Prolagen® inhalant SPT reagents are not inferior to the commercially available SPT reagents in allergy diagnosis.
Allergens ; Allergy and Immunology ; Ambrosia ; Animals ; Artemisia ; Cats ; Dermatophagoides farinae ; Dermatophagoides pteronyssinus ; Diagnosis ; Dogs ; Electrophoresis ; Electrophoresis, Polyacrylamide Gel ; Humans ; Humulus ; Hypersensitivity ; Immunoblotting ; Immunoglobulin E ; Immunoglobulins ; In Vitro Techniques* ; Indicators and Reagents* ; Korea ; Methods* ; Pollen ; Skin* ; Sodium

PURPOSE: Japanese hop (Humulus japonicus) is a major cause of weed pollinosis in East Asia. However, supplies of commercial allergen extract from this plant have not met clinical demand. The pollen of common hop (Humulus lupulus), a closely related species, may provide an alternative source if there is strong IgE cross-reactivity between these two species. We aimed to compare the IgE cross-reactivity and allergenicity of common hop and Japanese hop pollen. MATERIALS AND METHODS: Cross-reactivity was measured by inhibition ELISA. One- and two-dimensional (2D) gel analyses combined with IgE immunoblotting and mass spectrometry [liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS)] were performed to detect IgE-reactive pollen components. RESULTS: Up to 16.7% of IgE reactivity to Japanese hop was inhibited by common hop. A 12-kDa protein component of Japanese hop pollen that showed the most potent IgE reaction was absent from common hop. Six IgE-reactive components from Japanese hop were detected by 2D gel electrophoresis and LC-ESI-MS/MS, but showed low Mascot scores, preventing positive identification. CONCLUSION: No significant IgE cross-reaction was observed for Japanese and common hop pollen allergens. Development of allergy diagnostic and immunotherapeutic reagents based on Japanese hop pollen are urgently needed.
Allergens ; Asian Continental Ancestry Group ; Chromatography ; Electrophoresis, Gel, Two-Dimensional ; Enzyme-Linked Immunosorbent Assay ; Equipment and Supplies ; Far East ; Humans ; Humulus* ; Hypersensitivity ; Immunoblotting ; Immunoglobulin E* ; Indicators and Reagents ; Mass Spectrometry ; Plants ; Pollen ; Rhinitis, Allergic, Seasonal ; Tandem Mass Spectrometry

PURPOSE: The AdvanSure™ AlloScreen assay is an advanced multiplex test that allows for simultaneous detection of specific IgE (sIgE) against multiple allergens. For precise identification of causative allergens in allergic patients, we compared this new multiplex sIgE assay with the ImmunoCAP assay, which is currently the gold-standard method for sIgE detection. MATERIALS AND METHODS: Serum samples from 218 Korean allergic disease patients were used to compare the ImmunoCAP and AlloScreen assays with respect to the following 13 allergens: Dermatophagoides pteronyssinus, Dermatophagoides farinae, cat and dog dander, Alternaria, birch, oak, ragweed, mugwort, rye grass, and food allergens (egg white, cow's milk, peanuts). RESULTS: A total of 957 paired tests using the 13 allergens were compared. The total agreement ratio ranged from 0.74 (oak) to 0.97 (Alternaria). With respect to class association analyses, the gamma index ranged from 0.819 (rye grass) to 0.990 (Alternaria). The intra-class correlation coefficients for house dust mites, cat and dog dander, Alternaria, birch, ragweed, egg white, cow's milk, and peanut sIgE titers were >0.8. CONCLUSION: The AlloScreen and ImmunoCAP assays exhibited similar diagnostic performance. However, due to methodological differences between the two systems, careful interpretation of their results is needed in clinical applications.
Allergens ; Alternaria ; Ambrosia ; Animals ; Arachis ; Artemisia ; Betula ; Cats ; Dander ; Dermatophagoides farinae ; Dermatophagoides pteronyssinus ; Dogs ; Egg White ; Humans ; Immunoassay ; Immunoglobulin E* ; Lolium ; Methods ; Milk ; Pyroglyphidae

PURPOSE: Hypotension after emergency endotracheal intubation (ETI) is one of the major complications from emergency airway management. The aim of this study was to determine the possible risk factors that may predict postintubation hypotension (PIH) and its impact on in-hospital mortality. METHODS: We conducted a retrospective, standardized chart review of consecutive emergency department patients that required intubation between January 2011 and December 2014. Patients were divided into 2 groups according to the presence or absence of PIH. PIH was defined as any recorded systolic blood pressure with less than 90 mmHg or mean arterial pressure with less than 65 mmHg within the 60-minute period after intubation. The outcome measures were inhospital mortality, as well as intensive care unit and hospital length of stay. RESULTS: The incidence of PIH was 23% (80 of 352 patients). Patients in the PIH group were slightly older and had more comorbid diseases than those in the non-PIH group. PIH patients had a significantly higher mortality rate (54% vs. 30%, p<0.01). PIH was a strong predictor for in-hospital mortality of intubated patients (hazard ratio, 2.3; 95% confidence interval, 1.3 to 3.4). CONCLUSION: Older age, lack of skill, history of hypertension, low albumin and pH, and elevated were risk factors for the occurrence of hypotension after ETI. Patients with PIH show increased risk of in-hospital mortality.
Airway Management ; Arterial Pressure ; Blood Pressure ; Emergencies ; Emergency Service, Hospital ; Hospital Mortality ; Humans ; Hydrogen-Ion Concentration ; Hypertension ; Hypotension ; Incidence ; Intensive Care Units ; Intubation ; Intubation, Intratracheal* ; Length of Stay ; Mortality ; Outcome Assessment (Health Care) ; Retrospective Studies ; Risk Factors

BACKGROUND: In a rapidly aging population, the importance of the quality of life of the elderly has been emphasized. The need for special care services according to an individual's physical, emotional, and social functions changes with aging. This study was conducted to analyze the needs of home dwellers in terms of health-care services for active aging. METHODS: The study subjects were 869 adults older than 40 years who lived in houses. They responded to a survey that inquired about the importance of each health-care item for active aging at home. The important items for active aging at home were graded by using a 5-point Likert scale. RESULTS: Among the items, "affection" topped the list, with 4.68 points, followed by cognitive function (4.67 points) and "social relations" (4.59 points). The top 3 items had no significant differences between the age groups. However, some items related to activities of daily living (ADL; including bathing, eating, and ambulating.) and instrumental ADL (IADL; including taking medicines and preparing meals) had significant differences between the age groups. Compared with the subjects in the older age groups, the subjects in the forties age group placed less importance on ADL and IADL. CONCLUSION: For the subjects who lived in houses, affection, cognitive function, and social relation are highly important for as active aging, regardless of age. ADL and IADL are regarded as more important in ≥50 years age groups.
Activities of Daily Living ; Adult ; Aged ; Aging* ; Baths ; Delivery of Health Care* ; Eating ; Humans ; Quality of Life

We present a new next-generation sequencing-based method to identify somatic mutations of lung cancer. It is a comprehensive mutation profiling protocol to detect somatic mutations in 30 genes found frequently in lung adenocarcinoma. The total length of the target regions is 107 kb, and a capture assay was designed to cover 99% of it. This method exhibited about 97% mean coverage at 30x sequencing depth and 42% average specificity when sequencing of more than 3.25 Gb was carried out for the normal sample. We discovered 513 variations from targeted exome sequencing of lung cancer cells, which is 3.9-fold higher than in the normal sample. The variations in cancer cells included previously reported somatic mutations in the COSMIC database, such as variations in TP53, KRAS, and STK11 of sample H-23 and in EGFR of sample H-1650, especially with more than 1,000x coverage. Among the somatic mutations, up to 91% of single nucleotide polymorphisms from the two cancer samples were validated by DNA microarray-based genotyping. Our results demonstrated the feasibility of high-throughput mutation profiling with lung adenocarcinoma samples, and the profiling method can be used as a robust and effective protocol for somatic variant screening.
Adenocarcinoma ; DNA ; Exome ; High-Throughput Nucleotide Sequencing ; Lung ; Lung Neoplasms* ; Mass Screening ; Polymorphism, Single Nucleotide ; Sensitivity and Specificity

PURPOSE: Intravenous lipid emulsions have been used to treat the systemic toxicity of local anesthetics. The goal of this in vitro study was to examine the effects of lipid emulsions on the norepinephrine-mediated reversal of vasodilation induced by high doses of levobupivacaine, ropivacaine, and mepivacaine in isolated endothelium-denuded rat aorta, and to determine whether such effects are associated with the lipid solubility of local anesthetics. MATERIALS AND METHODS: The effects of lipid emulsions (0.30, 0.49, 1.40, and 2.61%) on norepinephrine concentration-responses in high-dose local anesthetic (6x10-4 M levobupivacaine, 2x10-3 M ropivacaine, and 7x10-3 M mepivacaine)-induced vasodilation of isolated aorta precontracted with 60 mM KCl were assessed. The effects of lipid emulsions on local anesthetic- and diltiazem-induced vasodilation in isolated aorta precontracted with phenylephrine were also assessed. RESULTS: Lipid emulsions (0.30%) enhanced norepinephrine-induced contraction in levobupivacaine-induced vasodilation, whereas 1.40 and 2.61% lipid emulsions enhanced norepinephrine-induced contraction in both ropivacaine- and mepivacaine-induced vasodilation, respectively. Lipid emulsions (0.20, 0.49 and 1.40%) inhibited vasodilation induced by levobupivacaine and ropivacaine, whereas 1.40 and 2.61% lipid emulsions slightly attenuated mepivacaine (3x10-3 M)-induced vasodilation. In addition, lipid emulsions attenuated diltiazem-induced vasodilation. Lipid emulsions enhanced norepinephrine-induced contraction in endothelium-denuded aorta without pretreatment with local anesthetics. CONCLUSION: Taken together, these results suggest that lipid emulsions enhance the norepinephrine-mediated reversal of local anesthetic-induced vasodilation at toxic anesthetic doses and inhibit local anesthetic-induced vasodilation in a manner correlated with the lipid solubility of a particular local anesthetic.
Amides/adverse effects ; Anesthetics, Local/*adverse effects ; Animals ; Bupivacaine/adverse effects/analogs & derivatives ; Emulsions/*chemistry/*therapeutic use ; Lipids/*chemistry ; Male ; Mepivacaine/adverse effects ; Norepinephrine/*therapeutic use ; Rats ; Rats, Sprague-Dawley ; Vasodilation/*drug effects