Clara cell protein in nasal lavage fluid and nasal nitric oxide - biomarkers with anti-inflammatory properties in allergic rhinitis

Background

Clara cell protein (CC16) is ascribed a protective and anti-inflammatory role in airway inflammation. Lower levels have been observed in asthmatic subjects as well as in subjects with intermittent allergic rhinitis than in healthy controls. Nasal nitric oxide (nNO) is present in high concentrations in the upper airways, and considered a biomarker with beneficial effects, due to inhibition of bacteria and viruses along with stimulation of ciliary motility. The aim of this study was to evaluate the presumed anti-inflammatory effects of nasal CC16 and nNO in subjects with allergic rhinitis.

Methods

The levels of CC16 in nasal lavage fluids, achieved from subjects with persistent allergic rhinitis (n = 13), intermittent allergic rhinitis in an allergen free interval (n = 5) and healthy controls (n = 7), were analyzed by Western blot. The levels of nNO were measured by the subtraction method using NIOX^?. The occurrences of effector cells in allergic inflammation, i.e. metachromatic cells (MC, mast cells and basophiles) and eosinophils (Eos) were analyzed by light microscopy in samples achieved by nasal brushing.

Results

The levels of CC16 correlated with nNO levels (r² = 0.37; p = 0.02) in allergic subjects. The levels of both biomarkers showed inverse relationships with MC occurrence, as higher levels of CC16 (p = 0.03) and nNO (p = 0.05) were found in allergic subjects with no demonstrable MC compared to the levels in subjects with demonstrable MC. Similar relationships, but not reaching significance, were observed between the CC16 and nNO levels and Eos occurrence. The levels of CC16 and nNO did not differ between the allergic and the control groups.

Conclusions

The correlation between nasal CC16 and nNO levels in patients with allergic rhinitis, along with an inverse relationship between their levels and the occurrences of MC in allergic inflammation, may indicate that both biomarkers have anti-inflammatory effects by suppression of cell recruitment. The mechanisms behind these observations warrant further analyses.     

Dynamic changes and clinical significance of serum tryptase levels in STEMI patients treated with primary PCI

Abstract

Objective: To investigate the dynamic changes in serum tryptase levels and their association with clinical data in patients with acute ST-elevation myocardial infarction (STEMI) treated with primary percutaneous coronary intervention (PCI).

Methods: Serum tryptase levels were measured in 99 STEMI patients and 25 control subjects.

Results: Tryptase levels were significantly increased at admission, and descended after primary PCI. Tryptase levels at 0.5, 2 and 6?h after PCI were negatively correlated with the percentage of ST-segment resolution (STR) as well as left ventricular ejection fraction (LVEF).

Conclusions: High tryptase levels after PCI were associated with poor myocardial reperfusion and poor cardiac function.     

Changes in glucose transporter expression and nitric oxide production are associated with liver injury in diabetes

In diabetes mellitus (DM), both hyperglycaemia and hyperlipidaemia can initiate accumulation of fat in the liver, which might be further mediated by inducible nitric oxide synthase. We have studied changes in GLUT1, nitric oxide (NO^·) concentration and liver damage in two rat DM models. STZ model was induced by strepozotocin 50 mg/kg. HS model was induced by high‐fat diet and 30 mg/kg streptozotocin. GLUT1 expression was studied by means of real‐time RT‐PCR and immunohistochemistry. Production of NO^· was monitored by means of erythrocyte sedimentation rate spectroscopy of Fe‐DETC‐NO complex. Liver damage was assessed using histological activity index (HAI). NO^· concentration was increased in the liver of STZ rats, but it did not change in HS rats (control 36.8 ± 10.3; STZ 142.1 ± 31.1; HS 35.4 ± 9.8 ng/g). Liver HAI was higher in STZ group, 8.6 ± 0.17 versus HS 4.7 ± 0.31, p < 0.05. GLUT1 protein expression was elevated only in STZ group, 16 ± 3 cells/mm² versus Control 5 ± 2 cells/mm², p = 0.007. Hyperglycaemia sooner causes severe liver damage in rat models of DM, compared with hyperlipidaemia, and is associated with increased NO^· production. GLUT1 transporter expression might be involved in toxic effects of glucose in the liver. We have obtained novel data about association of GLUT1 expression and NO^· metabolism in the pathogenesis of liver injury in DM. Increased GLUT1 expression was observed together with overproduction of NO^· and pronounced liver injury in severely hyperglycaemic rats. On the contrary, moderately hyperglycaemic hyperlipidaemic rats developed only moderate liver steatosis and no increase in GLUT1 and NO^·. GLUT1 overexpression might be implicated in the toxic effects of glucose in the liver. Glycotoxicity is associated with oxidative stress and NO^· hyperproduction. GLUT1 and NO^· metabolism might become novel therapeutic targets in liver steatosis. Copyright © 2015 John Wiley & Sons, Ltd.     

Interaction of caveolin-1, nitric oxide, and nitric oxide synthases in hypoxic human SK-N-MC neuroblastoma cells

Neuroblastoma cells are capable of hypoxic adaptation, but the mechanisms involved are not fully understood. We hypothesized that caveolin-1 (cav-1), a plasma membrane signal molecule, might play a role in protecting neuroblastoma cells from oxidative injury by modulating nitric oxide (NO) production. We investigated the alterations of cav-1, cav-2, nitric oxide synthases (NOS), and NO levels in human SK-N-MC neuroblastoma cells exposed to hypoxia with 2% [O2]. The major discoveries include: (i) cav-1 but not cav-2 was up-regulated in the cells exposed to 15 h of hypoxia; (ii) NO donor 1-[N, N-di-(2-aminoethyl) amino] diazen-1-ium-1, 2-diolate up-regulated the expression of cav-1, whereas the non-selective NOS inhibitor N(G)-nitro-L-arginine methyl ester and inducible NOS (iNOS) inhibitor 1400W each abolished the increase in cav-1 expression in the hypoxic SK-N-MC cells. These results suggest that iNOS-induced NO production contributes to the up-regulation of cav-1 in the hypoxic SK-N-MC cells. Furthermore, we studied the roles played by cav-1 in regulating NO, NOS, and apoptotic cell death in the SK-N-MC cells subjected to 15 h of hypoxic treatment. Both cav-1 transfection and cav-1 scaffolding domain peptide abolished the induction of iNOS, reduced the production of NO, and reduced the rates of apoptotic cell death in the hypoxic SK-N-MC cells. These results suggest that increased expression of cav-1 in response to hypoxic stimulation could prevent oxidative injury induced by reactive oxygen species. The interactions of cav-1, NO, and NOS could be an important signal pathway in protecting the neuroblastoma cells from oxidative injury, contributing to the hypoxic tolerance of neuroblastoma cells.     

Endothelial nitric oxide synthase genotype and haplotype are not associated with diabetic retinopathy in diabetes type 2 patients

Polymorphisms in the endothelial nitric oxide synthase (eNOS) gene have been associated with the development of diabetic retinopathy (DR) in patients with type 1 diabetes mellitus (T1DM), but not with T2DM. However, no previous study has analyzed combinations of genetic markers (haplotypes), which can be more informative. We studied three eNOS genetic polymorphisms: a single nucleotide polymorphism in the promoter region (T(-786)C), in exon 7 (Glu298Asp), and a variable number of tandem repeats in intron 4 (b/a) in 103 healthy controls, and in 170 patients with T2DM (without DR, N=114; with DR, N=56). We also examined the association of eNOS gene haplotypes with T2DM and with DR. No differences were found in the frequencies of genotypes and alleles of the three polymorphisms among the three groups of subjects. However, the "C-Glu-b" haplotype was more common in healthy controls (24%) than in T2DM patients (7%) (P=0.0001). Finally, no significant difference in the distribution of eNOS haplotypes frequencies was found when T2DM patients with or without DR were compared (P=0.7372). These findings suggest no association between DR and individual eNOS haplotypes in T2DM patients. The "C-Glu-b" haplotype, however, may have a protective effect against T2DM. Further studies should be conducted to address the molecular basis for such an effect.     

Superantigen-induced apoptotic death of tumor cells is mediated by cytotoxic lymphocytes, cytokines, and nitric oxide.

The bacterial superantigen staphylococcal enterotoxin A (SEA) is a potent inducer of CTL activity and cytokine production in vivo. Protein A (PA) of Staphylococcal aureus has been found to have diverse biological response modifying properties and to possess antitumor, antitoxic and antiparasitic effects. In this study we examined the anti-tumor effect of these two superantigens used separately as well as in combination in mice carrying the Ehrlich ascites tumor. With combined treatment, DNA cell cycle analysis of tumor cells showed a significant (P < 0.05) percentage of tumor cell death. Levels of the soluble mediators TNF-alpha, IFN-gamma and IL-1 as well as NO were elevated. Additionally, CD4(+) and CD8(+) specific T cells in spleen, thymus and PBMC in tumor carrying mice were increased (P < 0.01). Our data altogether suggests that enhanced tumor cell death is caused by the increased CTL activity, cytokine and nitric oxide levels, in response to the combined effect of SEA + PA.     

加味射干麻黄汤对重度支气管哮喘患者血清ECP、LPO、FeNO及肺功能的影响

目的:探讨加味射干麻黄汤对重度支气管哮喘患者血清嗜酸性粒细胞阳离子蛋白(ECP)、脂质过氧化物(LPO)、呼出气一氧化氮(FeNO)及肺功能的影响。方法:选择2015年9月至2017年9月我院接诊的96例重度支气管哮喘患者进行研究,通过随机数表法分为观察组(n=48)和对照组(n=48),在常规治疗基础上,对照组给予异丙托溴铵溶液、甲泼尼龙琥珀酸钠治疗,观察组联合加味射干麻黄汤治疗,均连续治疗4周。比较两组临床疗效、治疗前后中医证候积分、肺功能、血清ECP、LPO及FeNO的变化和不良反应。结果:治疗后,观察组临床疗效总有效率为91.67%(44/48),明显高于对照组的72.92%(35/48)(P0.05);治疗后,两组各中医证候积分、肺功能、血清ECP、LPO及FeNO较治疗前均显著改善(P0.05);中医证候积分中,观察组治疗后喘息、咳嗽、咳痰、胸闷、哮鸣音积分均明显低于对照组,肺功能中,观察组第1秒用力呼出量(FEV1)、FEV1/用力肺活量(FEV1/FVC)均明显高于对照组,呼气峰值流速(PEF)昼夜变异率明显低于对照组,且观察组血清ECP、LPO及FeNO明显比低于对照组,差异均具有统计学意义(P0.05);两组治疗期间不良反应比较无显著差异(P0.05)。结论:在重度支气管哮喘患者中应用加味射干麻黄汤效果显著,可有效改善肺功能及临床症状,其内在机制可能和调节气道慢性炎症反应、氧化应激反应,降低血清ECP、LOP及FeNO的表达相关,且用药安全性高,值得应用推广。     

Copper modulates activities of genistein, nitric oxide, and curcumin in breast tumor cells

Several papers have reported that low level of genistein (<8 microM), the major bioactive component of isoflavones, stimulates the growth of MCF-7 cells. In the present study, we found that genistein-induced growth stimulation of MCF-7 cells is inhibited in the presence of Cu(2+) (5 microM). Genistein induces the release of nitric oxide in MCF-7 cells in a concentration-dependent manner. The release of nitric oxide was inhibited by N(G)-nitro-L-arginine methyl ester, suggesting the possibility of the activation of nitric oxide synthase. The growth of MCF-7 cells also increases in the presence of low levels of sodium nitriprusside (<10 microM), a nitric oxide donor compound, while high levels (>25 microM) are toxic. The sodium nitroprusside-induced growth of MCF-7 cells is drastically suppressed in the presence of Cu(2+) (5 microM). This parallel behavior between Cu(2+)-genistein and Cu(2+)-sodium nitroprusside mixtures suggests that Cu(2+) and/or copper-protein complexes, that may be formed in the media, may be reacting with nitric oxide or nitric oxide-derived reactive species. The products of these reactions may be responsible for the toxic effects of these mixtures. In contrast, the effect of curcumin that inhibits the growth of both estrogen receptor-positive and -negative breast tumor cells appreciably decreased in the presence of Cu(2+). Since copper is known to overwhelmingly bind with proteins, present data suggest that an increase in copper-protein moieties or complexes formed in the serum containing media and their reactions with nitric oxide may be responsible for their toxic effects. Further studies are needed to characterize these reactions.     

Production of heat shock proteins, cytokines, and nitric oxide in toxic stress

Expression of heat shock proteins Hsp27, Hsp90, and Hsp70 and production of tumor necrosis factors (TNF-alpha, TNF-beta), interferon-gamma (IFN-gamma), interleukin-2, -3, -6, and nitric oxide (NO) were studied under conditions of acute and chronic intoxication of animals with lipopolysaccharides. Injection of endotoxin increased expression of heat shock proteins Hsp70 and Hsp90-alpha in mouse cells. Acute toxic stress also provoked a sharp increase in the production of TNF-alpha, TNF-beta, and NO in mouse cells. The production of other cytokines (interleukins and IFN-gamma) was changed insignificantly. In the model of chronic toxic stress, changes in the production of Hsp70, Hsp90, TNF, and NO were followed during 11 days after the beginning of the toxin injections. The expression of Hsp70 and Hsp90 in acute stress was significantly higher than at the final stage of the chronic exposure. The changes in the TNF and NO productions, on one hand, and the production of heat shock proteins, on the other hand, were synchronous. The findings indicate that repeated injections of increasing endotoxin doses result in a decreased ability of the body cells to respond to stress by overproduction of heat shock proteins, TNF, and NO.     

Cardiac malformations are associated with altered expression of vascular endothelial growth factor and endothelial nitric oxide synthase genes in embryos of diabetic mice

The aim of this study was to investigate the role of nitric oxide (NO), and the expression of endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) genes in developing hearts at embryonic day 13.5 of embryos from diabetic mice. The protein and mRNA expression levels of eNOS and VEGF were significantly altered in the developing hearts of embryos from diabetic mice. The NO level was significantly decreased, whereas the VEGF concentration was significantly increased in the developing hearts of the embryos from diabetic mice. In vitro study showed a significant reduction in eNOS expression and cell proliferation in cardiac myoblast cells exposed to high glucose concentrations. Further, high glucose induced apoptosis in myoblast cells. Ultrastructural changes characteristics of apoptosis, including cell blebbing, aggregation of ribosomes and vacuoles in the cytoplasm were also evident in myoblast cells exposed to high glucose. It is suggested that hyperglycemia alters the expression of eNOS and VEGF genes that are involved in the regulation of cell growth and vasculogenesis, thereby contributing to the cardiac malformations seen in embryos from diabetic mice.     

Lower uterine artery blood flow and higher endothelin relative to nitric oxide metabolite levels are associated with reductions in birth weight at high altitude

Reduced uteroplacental blood flow is hypothesized to play a key role in altitude-associated fetal growth restriction. It is unknown whether reduced blood flow is a cause or consequence of reduced fetal size. We asked whether determinants of uteroplacental blood flow were altered prior to reduced fetal growth and whether vasoactive and/or angiogenic factors were involved. Women residing at low (LA; 1,600 m, n = 18) or high altitude (HA; 3,100 m, n = 25) were studied during pregnancy (20, 30, and 36 wk) and 4 mo postpartum (PP) using Doppler ultrasound. In each study, endothelin (ET-1), nitric oxide metabolites (NO(x)), soluble fms-like tyrosine kinase (sFlt-1) and placental growth factor (PlGF) levels were quantified. At HA, birth weights were lower (P < 0.01) and small-for-gestational age was more common (P < 0.05) compared with LA. HA was associated with lower uterine artery (UA) diameter (P < 0.01) and blood flow (P < 0.05). Altitude did not affect ET-1, sFlt-1 or PlGF; however, ET-1/NO(x) was greater and NO(x) lower during pregnancy and PP at HA vs. LA. ET-1/NO(x) was negatively associated with birth weight (20 wk, P < 0.01; 36 wk, P = 0.05) at LA and HA combined. At HA, UA blood flow (30 wk) was positively associated with birth weight (dagger). UA blood flow and ET-1/NO(x) levels accounted for 45% (20 wk) and 32% (30 wk) of birth weight variation at LA and HA combined, primarily attributed to effects at HA. We concluded that elevated ET-1/NO(x) and altered determinants of uteroplacental blood flow occur prior to altitude-associated reductions in fetal growth, and therefore, they are likely a cause rather than a consequence of smaller fetal size.     

Infestation status of head louse and treatment with lindane shampoo in children of primary school and kindergarten in Chinju-shi, Kyongsangnam-do, Korea

The infestation status of head louse among children attending primary schools and kindergartens in Chinju-shi, Kyongsangnam-do, Korea, was investigated between June and July 1999. Out of 2,288 children examined, 3.9% of boys (48/1,242) and 23.5% of girls (246/1,046) were infested with nits or adult/nymphs of lice. The effectiveness of lindane shampoo (1% gamma benzene hexachloride solution) was evaluated after one or two time applications to all the children infested. The negative conversion rate of pediculosis was 93.5%. Effective control measures are needed to control and prevent such ectoparasite infestation amongst children.     

Risk factors for asthma and allergy associated with urban migration: background and methodology of a cross-sectional study in Afro-Ecuadorian school children in Northeastern Ecuador (Esmeraldas-SCAALA Study)

Background

This study examined the attitudes and actions of 3415 physician-recruited adults aged ≥ 16 years with asthma in eleven countries who were prescribed regular maintenance therapy with inhaled corticosteroids or inhaled corticosteroids plus long-acting β₂-agonists.

Methods

Structured interviews were conducted to assess medication use, asthma control, and patients' ability to recognise and self-manage worsening asthma.

Results

Despite being prescribed regular maintenance therapy, 74% of patients used short-acting β₂-agonists daily and 51% were classified by the Asthma Control Questionnaire as having uncontrolled asthma. Even patients with well-controlled asthma reported an average of 6 worsenings/year. The mean period from the onset to the peak symptoms of a worsening was 5.1 days. Although most patients recognised the early signs of worsenings, the most common response was to increase short-acting β₂-agonist use; inhaled corticosteroids were increased to a lesser extent at the peak of a worsening.

Conclusion

Previous studies of this nature have also reported considerable patient morbidity, but in those studies approximately three-quarters of patients were not receiving regular maintenance therapy and not all had a physician-confirmed diagnosis of asthma. This study shows that patients with asthma receiving regular maintenance therapy still have high levels of inadequately controlled asthma. The study also shows that patients recognise deteriorating asthma control and adjust their medication during episodes of worsening. However, they often adjust treatment in an inappropriate manner, which represents a window of missed opportunity.     

食物过敏患儿肠道菌群特征及其与外周血Treg/Th17的相关性

分析食物过敏患儿肠道菌群分布与外周血免疫细胞Treg/Th17的关系。

选择2020年4月至2022年5月本院儿科收治的86例食物过敏患儿作为观察组,另选择同期80例健康儿童作为健康对照组。采集两组儿童新鲜粪便和外周血样本,分别测定肠道菌群数量以及外周血Treg/Th17细胞、血清相关细胞因子水平,并经Pearson相关性分析肠道菌群与外周血Treg/Th17的相关性。

观察组患儿肠道内乳杆菌、双歧杆菌数量均低于健康对照组（均P＜0.05）;观察组患儿外周血内Treg细胞水平、Treg/Th17比值均低于健康对照组,但Th17细胞水平高于健康对照组（均P＜0.05）;观察组患儿的血清白细胞介素-17（interleukin-17,IL-17）水平高于健康对照组,血清转化生长因子-β1（transforming growth factor-β1,TGF-β1）指标低于健康对照组（均P＜0.05）;Pearson相关性分析结果显示,乳杆菌、双歧杆菌与外周血Treg细胞、TGF-β1水平均呈正相关,而与外周血Th17细胞、IL-17水平均呈负相关（均P＜0.05）。

食物过敏患儿伴显著的肠道菌群紊乱与免疫功能异常状况,肠道内乳杆菌和双歧杆菌水平以及外周血Treg细胞、TGF-β1表达减少,外周血Th17细胞、IL-17表达增多。临床应改善患儿肠道微生态环境,调控肠道菌群结构,促进肠道菌群分布平衡恢复。

     

Sialyl-Lewis x and Sialyl-Lewis a are associated with MUC1 in human endometrium

Endometrial epithelial cells express MUC1 with increased abundance in the secretory phase of the menstrual cycle, when embryo implantation occurs. MUC1 is associated with the apical surface of epithelial cells and is also secreted, being detectable in uterine fluid at elevated levels in the implantation phase. However, its physiological role is uncertain; it may either inhibit intercellular adhesion by steric hindrance or carry carbohydrate recognition structures capable of mediating cell-cell interaction. Here we show that endometrial epithelium expresses both Sialyl-Lewis x (SLe^x) and Sialyl-Lewis a (SLe^a), with a distribution and pattern of menstrual cycle regulation similar to that of MUC1. Using Western blotting and double determinant ELISA of uterine flushings, we demonstrate that SLe^x is associated with MUC1 core protein. The endometrial carcinoma cell lines HEC1A and HEC1B are shown to express MUC1 in a mosaic pattern, while three other cell lines express much lower amounts. HEC1A expresses both SLe^x and SLe^a while HEC1B expresses SLe^a only. Immunoprecipitation has been used to demonstrate that SLe^a is associated with MUC1 in HEC1B cells, and both SLe^x and SLe^a are associated with MUC1 in HEC1A cells.     

Induction of iron regulatory protein 1 RNA-binding activity by nitric oxide is associated with a concomitant increase in the labile iron pool: implications for DNA damage

Iron regulatory protein 1 (IRP1) is a bifunctional [4Fe-4S] protein that controls iron homeostasis. Switching off its function from an aconitase to an apo-IRP1 interacting with iron-responsive element-containing mRNAs depends on the reduced availability of iron in labile iron pool (LIP). Although the modulation of IRP1 by nitric oxide has been characterized, its impact on LIP remains unknown. Here, we show that inhibition of IRP1 aconitase activity and induction of its IRE-binding activity during exposure of L5178Y mouse lymphoma cells to NO are associated with an increase in LIP levels. Removal of NO resulted in a reverse regulation of IRP1 activities accompanied by a decrease of LIP. The increased iron burden in LIP caused by NO exacerbated hydrogen peroxide-induced genotoxicity in L5178Y cells. We demonstrate that the increase in LIP levels in response to chronic but not burst exposure of L5178Y cells to NO is associated with alterations in the expression of proteins involved in iron metabolism.     

Upregulation of neuronal nitric oxide synthase, edema and cell injury following heat stress are reduced by pretreatment with EGB-761 in the rat

Rats exposed to 4 h heat stress (HS) at 38°C exhibited marked upregulation of neuronal nitric oxide synthase (nNOS) in the brain regions exhibiting blood–brain barrier (BBB) breakdown, brain edema and cell damage. Pretreatment with an anti-oxidant compound EGB-761 (an extract of Gingko biloba) administered 50 mg/kg, per os for 5 days, significantly attenuated nNOS expression, BBB disruption, brain edema and cell injury. These results suggest that EGB-761 is neuroprotective in heat stress and this effect of the compound is related with the inhibition of NOS expression, not reported earlier.     

Functions and dysfunctions of nitric oxide in brain

Nitric oxide (NO) works as a retrograde neurotransmitter in synapses, allows the brain blood flow and also has important roles in intracellular signaling in neurons from the regulation of the neuronal metabolic status to the dendritic spine growth. Moreover NO is able to perform post-translational modifications in proteins by the S-nitrosylation of the thiol amino acids, which is a physiological mechanism to regulate protein function. On the other hand, during aging and pathological processes the behavior of NO can turn harmful when reacts with superoxide anion to form peroxynitrite. This gaseous compound can diffuse easily throughout the neuronal membranes damaging lipid, proteins and nucleic acids. In the case of proteins, peroxynitrite reacts mostly with the phenolic ring of the tyrosines forming nitro-tyrosines that affects dramatically to the physiological functions of the proteins. Protein nitrotyrosination is an irreversible process that also yields to the accumulation of the modified proteins contributing to the onset and progression of neurodegenerative processes such as Alzheimer's disease or Parkinson's disease.