Role and regulation of SPLUNC1 in respiratory tract infection / 国际儿科学杂志

Respiratory tract and the external environment are interlinked,long-term exposure to a variety of physical and chemical substances and pathogenic microorganism stimulation,the antimicrobial activity of proteins secreted by epithelial cells is critical for maintaining health.short palate,lung,and nasal epithelium clone 1 (SPLUNC1) is a lately discovered protein with antibacterial activity.Its structure is similar to the bactericidal permeability-increasing protein(BPI).SPLUNC1 binds specifically to lipopolysaccharide(LPS) of cell walls of gram-negative bacteria,and contributes to maintain homeostasis and a sterile environment in the lung,and acts as a goalkeeper role in the rapid activation of innate immunity and the initiation of adaptive immunity.SPLUNC1 is expected to become an antimicrobial agents for the treatment of respiratory tract bacterial infectious diseases.

Genotype analysis of Acanthamoeba isolated from human nasal swabs in the Philippines

Objective: To analyze the genotypes of Acanthamoeba species isolated from human nasal swabs in the Philippines. Methods: Human nasal swabs were collected from two groups: a low exposure group composed of students of the University of the Philippines-Diliman and a high exposure group composed of laborers frequently exposed to garbage, soil and dust. After isolation using non-nutrient agar plate lawned with Escherichia coli and DNA extraction using Chelex-100 resin, the ASA.S1 region of the gene (Rns) coding for nuclear, small subunit ribosomal RNA of Acanthamoeba was amplified through polymerase chain reaction. Purified polymerase chain reaction products were then sequenced. Neighbor-joining, maximum parsimony, and maximum likelihood phylogenetic trees were then constructed. Results: In the low exposure group, 1 out of 70 (1.43%) students and 7 out of 110 (6.36%) in the high exposure group were culture-positive. Four soil samples were also obtained for comparison, all of which were tested culture-positive. Of the 12 Acanthamoeba isolates, only 9 were successfully sequenced. The basic local alignment search tool of the US National Center for Biotechnology Information was used to identify most similar sequences. Five isolates were identified as genotype T5, and 3 isolateds were genotype T4. Genotype T11 was also isolated from soil, the first to be reported in the Philippines. Conclusions: Genotype T11 is a possible pathogenic strain and both T4 and T5 can be pathogenic to human, hence, healthy provisions, especially for high exposure groups, should be given more attention and reevaluated.

Genotype analysis of Acanthamoeba isolated from human nasal swabs in the Philippines

OBJECTIVE@#To analyze the genotypes of Acanthamoeba species isolated from human nasal swabs in the Philippines.@*METHODS@#Human nasal swabs were collected from two groups: a low exposure group composed of students of the University of the Philippines-Diliman and a high exposure group composed of laborers frequently exposed to garbage, soil and dust. After isolation using non-nutrient agar plate lawned with Escherichia coli and DNA extraction using Chelex-100 resin, the ASA.S1 region of the gene (Rns) coding for nuclear, small subunit ribosomal RNA of Acanthamoeba was amplified through polymerase chain reaction. Purified polymerase chain reaction products were then sequenced. Neighbor-joining, maximum parsimony, and maximum likelihood phylogenetic trees were then constructed.@*RESULTS@#In the low exposure group, 1 out of 70 (1.43%) students and 7 out of 110 (6.36%) in the high exposure group were culture-positive. Four soil samples were also obtained for comparison, all of which were tested culture-positive. Of the 12 Acanthamoeba isolates, only 9 were successfully sequenced. The basic local alignment search tool of the US National Center for Biotechnology Information was used to identify most similar sequences. Five isolates were identified as genotype T5, and 3 isolateds were genotype T4. Genotype T11 was also isolated from soil, the first to be reported in the Philippines.@*CONCLUSIONS@#Genotype T11 is a possible pathogenic strain and both T4 and T5 can be pathogenic to human, hence, healthy provisions, especially for high exposure groups, should be given more attention and reevaluated.

Efeitos da participação de esteroides-like provenientes da poluição atmosférica no epitélio das vias aéreas em camundongos machos e fêmeas / Effects of the participation of steroid-like compounds from air pollution in the airway epithelium of male and female mice

O epitélio nasal é a primeira porção do sistema respiratório a entrar em contato com o ambiente externo. Partículas da poluição do ar, principalmente os compostos orgânicos absorvidos, podem atuar como liberadores endócrinos. O receptor aril hidrocarboneto (AhR) é um importante competidor dos receptores de estrógeno-beta (ERbeta) que regulam a transcrição do gene para enzimas de metabolização xenobióticas (enzimas do citocromo P450). O objetivo deste estudo é identificar e quantificar ERbeta, AhR, CYP1A1, CYP1A2, CYP1B1 e o perfil de muco no epitélio nasal de camundongos machos e fêmeas em diferentes fases do ciclo estral. Camundongos BALB/c machos (n=32) e fêmeas (n=84) foram expostos ao ar ambiente e ao MP2,5 concentrado a 600 ug.m-³ em um concentrador de partículas ambientais (CPAs). As fêmeas foram divididas de acordo com as fases do ciclo estral: proestro, estro e diestro. O epitélio nasal foi avaliado por RT-PCR e imuno-histoquímica para análise de expressão de ERbeta (proteína), Erbeta-1 e Erbeta-2 (gene), AhR (proteína e gene) e Cyp1a1, Cyp1a2 and Cyp1b1 (gene). A quantificação de muco neutro - Periodic Acid Schiff's (PAS+) e ácido - Alcian Blue (AB+) foi avaliada por morfometria. As exposições foram realizadas durante 5 dias/semana, por 45 ± 55 dias. A expressão de Erbeta-2 RNAm apresentou diferenças em resposta à exposição ao CPAs (p=0,016), bem como uma diminuição em fêmeas, quando comparadas aos camundongos machos (p=0,036). A expressão de Cyp1b1 RNAm foi significantemente menor no grupo exposto ao CPAs, em relação ao grupo exposto ao ar ambiente nas fêmeas em diestro (p=0,036). A expressão de Erbeta foi aumentada no epitélio nasal de fêmeas em estro expostas ao CPAs (p=0,005) e a expressão de AhR foi menor em fêmeas em proestro expostas ao CPAs (p=0,048). A exposição ao CPAs levou ao aumento do conteúdo de muco ácido em camundongos machos (p=0,048), o qual diminuiu em fêmeas (p=0,040), quando comparados ao grupo ar ambiente. Este estudo mostrou...

The nasal epithelium is the first portion of the respiratory system to reach contact with the external environment. Air pollution particles, mainly the organic compounds absorbed into them, may act as endocrine releasers. The aryl hydrocarbon (AhR) receptor is an important competitor of estrogenic receptors-beta (ERbeta) that regulate transcription of gene coding for xenobiotic-metabolizing enzymes (cytochrome P450 enzymes). The aim of this study is to identify and quantify in the nasal epithelium of male and female mice in different estrous cycle phases related with ERbeta, AhR, CYP1A1, 1A2, 1B1 and the mucus profile. Male (n=32) and female (n=84) BALB/c mice were exposed to ambient air and PM2.5 concentrated at 600 ug.m-³ in an ambient particle concentrator with a particulate matter diameter of 2.5 um (PM2.5). Females were subdivided in three estrous cycles: proestrus, estrus and diestrus. Nasal epithelium was evaluated through RT-PCR and immunohistochemistry for the expression of ERbeta (protein), Erbeta-1 and Erbeta-2 (gene expression), AhR (protein and gene expression) and Cyp1a1, Cyp1a2 and Cyp1b1 (gene expression). Morphometry was applied for evaluation of mucus profile: acid - Alcian Blue (AB+) and neutral - Periodic Acid Schiff's (PAS+). Exposure happened for 5 days/week, for 45 ± 55 days. There were differences in Erbeta-2 mRNA in response to exposition to CPAs (p=0.016), and a significant decrease in female compared male mice (p=0.036). Cyp1b1 mRNA was significantly smaller in the CPAs-exposed group compared with the ambient air group in diestrus female mice (p=0.036). The ERbeta expression increased in the nasal epithelium of CPAs-exposed females in the estrus cycle (p=0.005), and the AhR expression decreased in the proestrus cycle of CPAs-exposed females (p=0.048). The exposure to the CPAs led to an increase in the acidic content of mucus in male mice (p=0.048), and decreased in female mice (p=0.040), compared to the ambient air group. This...

Expression of Bitter Taste Receptors in Human Nasal Respiratory Epithelium

The nasal cavity encounters various irritants during inhalation such as dust and pathogens. To detect and remove these irritants, it has been postulated that the nasal mucosa epithelium has a specialized sensing system. The oral cavity, on the other hand, is known to have bitter taste receptors (T2Rs) that can detect harmful substances to prevent ingestion. Recently, solitary chemosensory cells expressing T2R subtypes have been found in the respiratory epithelium of rodents. In addition, T2Rs have been identified in the human airway epithelia. However, it is not clear which T2Rs are expressed in the human nasal mucosa epithelium and whether they mediate the removal of foreign materials through increased cilia movement. In our current study, we show that human T2R receptors indeed function also in the nasal mucosa epithelium. Our RT-PCR data indicate that the T2R subtypes (T2R3, T2R4, T2R5, T2R10, T2R13, T2R14, T2R39, T2R43, T2R44, T2R 45, T2R46, T2R47, T2R48, T2R49, and T2R50) are expressed in human nasal mucosa. Furthermore, we have found that T2R receptor activators such as bitter chemicals augments the ciliary beating frequency. Our results thus demonstrate that T2Rs are likely to function in the cleanup of inhaled dust and pathogens by increasing ciliary movement. This would suggest that T2Rs are feasible molecular targets for the development of novel treatment strategies for nasal infection and inflammation.

Interleukin-1beta Decreases Caveolin-1 Expression in Human Nasal Epithelium / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Caveolin-1 (Cav-1) is the structural protein that is necessary for the formation of caveolae membrane domains. It is known as an inhibitor of various signaling pathways and associated with several diseases such as cancer, atherosclerosis, restrictive lung disease and obesity. However, studies for Cav-1 in nose has been hardly performed. The objectives of our study were to detect Cav-1 expression in human nasal epithelium and to investigate the change of Cav-1 expression in the inflammation of nasal epithelium. SUBJECTS AND METHOD: We obtained nasal polyp specimens from three patients undergoing endoscopic sinus surgery. Cells from specimens were cultured using the air-liquid interface technique and IL-1beta was treated. The expression of Cav-1 mRNA and protein was determined by reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis, respectively. RESULTS: Both RT-PCR and Western blot analysis demonstrated the presence of Cav-1 mRNA and protein in human nasal epithe-lium. Furthermore, the expression of both Cav-1 mRNA and protein was decreased by IL-1beta stimulation. CONCLUSION: Cav-1 was expressed in human nasal epithelial cells. It is assumed that Cav-1 may play a role in nasal inflammatory disease. However, further studies to confirm the interaction between Cav-1 and signaling molecules in the nasal inflammatory process should be followed.

Mucin and Lysozyme Expression in the Epithelium of Normal Human Nasal Inferior Turbinate and Polyps / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: In chronic bronchitis, rhinitis or cystic fibrosis, the number of goblet cells increases along with hypertrophy of mucous cells in submucosal gland, resulting ineffective mucociliary clearance. But, it is still not fully understood what role each gene plays in producing airway secretions. This study aimed to figure out which mucin gene is expressed in the epithelium of normal human nasal mucosa and nasal polyps, and to verify whether the epithelium of nasal polyp itself contributes to the increased nasal secretion as in chronic sinusitis with nasal polyp. MATERIALS AND METHODS: Normal nasal epithelial cells were obt assay. And, RT-PCR was used for the detection of mucin mRNA and lysozyme mRNA. RESULTS: The level of intracellular mucin was 2.9 times higher in the epithelium of nasal polyp, and this was statistically significant. Among seven mucin genes (MUC1, 2, 4, 5AC, 5B, 7, 8) expressed in the epithelium of normal inferior turbinate and polyps, MUC2 and MUC8 were more strongly expressed in the epithelium of nasal polyp than those of normal inferior turbinate. CONCLUSION: This results suggest that the polyp epithelium itself is contributing to increased secretion in chronic sinusitis, and MUC2 and MUC8 are thought tbe responsible for this change. However, further study is required to uncover the full sequence of MUC8 mRNA and its exact function.

Distribution of IL-4, IL-8 and GM-CSF in Nasal Turbinate Mucosa Following Sulfur Dioxide Exposure In Vitro

There is increasing evidence that airway epithelial cells, when exposed to various gas-derived air pollutants, play an important role in airway inflammation by releasing inflammatory cytokines. However, there is little information on air pollutant-induced cytokine expression at the tissue level and on the role of sulfur dioxide (SO2), one of the major ambient air pollutants, in cytokine production. We studied whether or not a low concentration of sulfur dioxide induces an increase in tissue expression of interleukin-4 (IL-4), interleukin-8 (IL-8), and granulocyte/macrophage colony stimulating factor (GM-CSF). After exposing surgically obtained normal human nasal turbinates to 0.05 ppm SO2 for one hour, we conducted specific immunohistochemical staining to assess the tissue expression of each cytokine. We found that the percent expression of IL-8 and GM-CSF in the surface epithelium was significantly higher in each SO2-exposed tissue than in the matched control tissue. However, there was no significant difference in the number of submucosal IL-4-positive cells between exposed and control specimens. These results suggest that exposure to a low concentration of SO2 increases airway inflammation, apparently by inducing an increase in the expression of GM-CSF and IL-8.

Expression of Inducible Nitric Oxide Synthase in Rat and Guinea PigRespiratory Epithelium after Capsaicin Treatment / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Nitric oxide (NO) production in the respiratory epithelium and the demonstration of inducible nitric oxide synthase in ciliated epithelium of the upper airway have recently been reported. The aim of this study was to investigate the expression of inducible nitric oxide synthase in the nasal epithelium after capsaicin treatment, which stimulates the substance P innervation. MATERIALS AND METHODS: In vivo treatment -Capsaicin (112 nM) was applied to the nasal cavities of the rat and guinea pig, and 30 nl of normal saline was applied for the control groups. After 2 hours, animals were sacrificed with cardiac perfusion of 4% paraformaldehyde and septal mucosa were removed. The 8 nm serial frozen tissue sections were made, and the expression of inducible nitric oxide synthase was determined using nicotinamide adenine diphosphate-diaphorase histochemistry. In vitro treatment- The nasal septum of the rats and the trachea of the guinea pigs were incubated in DMEM culture media with or without 112 nM capsaicin for experimental or control groups. After 0, 30 or 120 minutes of incubation, the tissues were fixed and processed for nicotinamide adenine diphosphate-diaphorase histochemistry. RESULTS: Both in vivo and in vitro studies demonstrated that the strong positive histochemical reactivity were observed in the respiratory epithelium of the rats and guinea pigs after capsaicin treatment compared to control groups. CONCLUSION: These data imply that capsaicin induces the expression of inducible nitric oxide synthase and that the substance P innervation of the nasal mucosa may have a protective role in the airway defense mechanism through nitric oxide production.