Resident mast cells are important for eotaxin-induced eosinophil accumulation in vivo

Cryosurgery may be considered for patients whose hepatic lesions are not amenable to surgical resection, i.e., patients with multiple hepatic lesions and/or lesions abutting major vascular structures. Because the size of the iceball created during the procedure can be carefully controlled, cryosurgery has the advantage of being a focal technique that spares much more noncancerous liver tissue than surgical resection. The major complications of hepatic cryosurgery are the same as those of hepatic resection: hemorrhage, pleural effusion, bile leak fistula, perihepatic abscess, and hepatic failure. In addition, there is a risk of coagulopathy when large tumors are frozen using multiple freeze-thaw cycles. In general, operative morbidity is related to the volume of frozen tissue, the number of freeze-thaw cycles, and number of cryoprobes. Further experience and accrual of long-term data should better define the indications for hepatic cryosurgery and minimize the incidence of complications.     

Effect of fluticasone propionate on sputum of patients with chronic bronchitis and emphysema

The effects of fluticasone propionate (FP) on sputum chemotactic activity, elastase inhibitory potential, albumin concentrations, and peripheral neutrophil function were studied in a group of patients with clinically stable, smoking-related chronic bronchitis and emphysema. Seventeen patients (50 to 75 yr of age) were entered into a double-blind, placebo-controlled study of 1.5 mg inhaled FP/d for 8 wk. Following treatment with FP the chemotactic activity of the sputum sol phase was lower than the corresponding values for the placebo group (p < 0.01). Values fell from a mean of 21.75 (+/- 1.58) during the run-in period to 18.37 (+/- 1.46; p < 0.01) after 4 wk and 17.63 (+/- 1.86; p < 0.05) after 8 wk treatment returning to 22.08 (+/- 1.26) cell/field after the washout period. The neutrophil elastase inhibitory capacity of the sputum sol phase increased (p < 0.025) with treatment from a mean of 0.177 microM elastase inhibited/L (+/- 0.05) pretreatment to 0.413 microM (+/- 0.054) after 4 wk and 0.415 microM (+/- 0.054) after 8 wk returning to 0.270 microM (+/- 0.07) after the washout period. Treatment with FP did not result in a change in the peripheral neutrophil functions studied or sputum albumin and myeloperoxidase concentrations. The results suggest that FP may play a protective role in these patients through a reduction in the chemotactic activity of lung secretions and potentially a reduction in the recruitment of neutrophils to the lung, and also by directly affecting the proteinase/antiproteinase balance, in favor of antiproteinases, within lung secretions.     

Induced sputum eosinophil cationic protein (ECP) measurement in asthma and chronic obstructive airway disease (COAD)

BACKGROUND: Induced sputum is a useful way to monitor airway inflammation in asthma, but cell counts are time-consuming and labour intensive. OBJECTIVE: The aim of this study was to evaluate a novel processing method using eosinophil cationic protein (ECP) as a biochemical marker of sputum eosinophil number and activation in subjects with asthma and other airway diseases. METHODS: Sputum was dispersed with dithiothreitol and centrifuged to yield cell free supernatant and a cell pellet. The pellet was treated with a cellular lysis buffer to release cell-associated ECP. ECP was measured in sputum supernatant and in the lysed cell pellet and was compared with sputum eosinophil counts in 31 adults with asthma, chronic obstructive airway disease (COAD), bronchiectasis and healthy controls. The ratio of supernatant to pellet ECP was evaluated as an index of eosinophil degranulation. The effect of sputum processing reagents and storage time on ECP measurement was also evaluated. RESULTS: ECP measured in the cell pellet lysate correlated closely with sputum absolute eosinophil counts across a range of subject groups (r = 0.72, P = 0.004). Sputum eosinophil counts were less well correlated with supernatant ECP levels (r = 0.54, P < 0.05). Incubation with dithiothreitol or lysis buffer did not influence ECP measurement and sputum ECP levels were stable over a 6-9 month period. Sputum supernatant and pellet lysate ECP concentrations were increased in stable asthma, asthma exacerbations and COAD/bronchiectasis (P < 0.05). The ratio of supernatant to pellet ECP was used as an index of eosinophil degranulation and found to be elevated in asthma exacerbations, COAD and bronchiectasis, but not in stable asthma. CONCLUSION: The measurement of ECP in the sputum cell pellet provides a reliable and efficient estimate of sputum eosinophil counts which can potentially be used in clinical trials and epidemiological surveys. The ECP ratio may be a useful marker of eosinophil activation, and was increased in asthma exacerbation and COAD. The increased ECP in COAD reflects a non-selective accumulation of eosinophils in this condition.     

Detection of K-ras point mutations in sputum from patients with adenocarcinoma of the lung by point-EXACCT

PURPOSE: Kirsten ras (K-ras) point mutations are found in 30% to 56% of pulmonary adenocarcinomas by means of highly sensitive techniques. Recently, the Point-EXACCT (point mutation detection using exonuclease amplification coupled capture technique) method was described, which detected one cell with a mutation in 15,000 normal cells. The aim of this study was to examine whether K-ras point mutations could be found with this rapid method in the sputum of patients with adenocarcinoma of the lung. PATIENTS AND METHODS: DNA from paraffin-embedded adenocarcinoma and corresponding sputum samples were analyzed for mutations of the K-ras gene. Twenty-eight biopsy specimens and 54 sputum samples of 22 patients were used for amplification and K-ras codon 12 point mutation detection. RESULTS: In 11 of 22 patients (50%), a mutation in K-ras codon 12 was shown in the tumor sample. In five of 11 patients (45%) with a K-ras mutation in the tumor, the same type of mutation was identified in at least one sputum sample. A mutation could not be detected in any of the sputum samples from patients with a K-ras-negative tumor. Time between K-ras point mutation detection in sputum and clinical diagnosis of lung cancer varied from 1 month to almost 4 years. In two of the five patients with K-ras-positive sputum specimens, malignant cells were found with cytologic examination. CONCLUSION: Point-EXACCT is suitable for the detection of K-ras point mutations in sputum samples of patients with adenocarcinoma of the lung. This approach may be an important adjunct to cytology in the early diagnosis of lung cancer.     

The relationship of eosinophil viability enhancing activity in sputum and clinical symptoms in asthma

We studied the correlation of eosinophil viability enhancing activity (EVEA) in sputum from asthmatic children and clinical symptoms. Sputum from asthmatic children and equal volume of saline were mixed with a Vortex mixer and centrifuged at 40000 G. Clear supernatants were obtained and filtered with 0.22 micron membrane filter. Periphral blood eosinophils purified by Percoll density gradient centrifugation and CD16 negative selection/ immunomagnetic beads technique were incubated with sputum extract for 4 days. Eosinophil viability was examined by staining the cells with fluorescen diacetate and propidium iodide and expressed as eosinophil viability enhancing activity (EVEA). Eosinophil cationic protein (ECP) concentrations in sputum were also measured by a radioimmunoassay. Correlation between EVEA in sputum and pulmonary functions, attack score, treatment score, or sputum ECP was determined by Spearman correlation test. Sputum EVEA was correlated 1) inversely with pulmonary functions on sampling, 2) with attack score of 2 weeks period before sampling, 3) with treatment score of 1 month to 1 week before and 1 to 2 weeks after sampling, and 4) with sputum ECP in individual cases. Sputum EVEA may serve as a suitable parameter for monitoring airway inflammation in asthma.     

Increased iron and ferritin content of sputum from patients with cystic fibrosis or chronic bronchitis

PURPOSE: Extracellular free iron, or iron bound to ferritin, may promote oxidative injury and bacterial growth in airways of patients with chronic airway inflammation due to cystic fibrosis (CF) or chronic bronchitis (CB). In this study, we assessed sputum content of total iron, ferritin, and transferrin in patients with CF or CB as well as sputum from normal subjects with acute airway inflammation caused by viral upper respiratory tract infections (URTIs). METHODS: Spontaneously produced sputum was obtained from 33 subjects, including 10 subjects with CF, 18 subjects with CB (10 acute exacerbations, 8 with stable CB), and 5 subjects with URTIs (control subjects). After lysing and dilution, total iron concentrations were determined by controlled coulometry, ferritin was measured by radioimmunoassay, and transferrin was measured by enzyme-linked immunosorbent assay. RESULTS: Iron was not present in detectable amounts in control sputums, but ferritin was present (6+/-2 ng/mg protein, mean+/-SE), as was transferrin (2.37+/-0.44 microg/mg). Compared with control subjects, concentrations of iron in sputum were increased in patient groups with higher amounts in CF patients (242+/-47 ng/mg, p<0.01) than CB patients with acute exacerbations or patients with stable CB (98+/-50 and 42+/-12 ng/mg, p<0.05 for both). Ferritin content of sputum was also increased in each group, with CF patients (113+/-22 ng/mg, p<0.001) higher than CB patients (acute, 45+/-10 ng/mg; stable, 87+/-24 ng/mg; p<0.01 for both). Compared with control subjects, sputum transferrin was decreased in CF patients (1.09+/-0.40 microg/mg, p<0.05), but not CB patients. CONCLUSIONS: These findings indicate there are increased airway concentrations of total iron and ferritin-bound iron in patients with CB and, to a greater extent, in patients with CF. Particularly in CF patients who also demonstrated decreased airway concentrations of transferrin, ferritin-bound iron in airways may promote oxidative injury and enhance bacterial growth.     

慢性阻塞性肺疾病患者气道过敏毒素C3a的变化

目的:研究补体裂解产物过敏毒素(C3a)在慢性阻塞性肺疾病(chronic obstruct1ve pulmonary disease,COPD)发病中的作用.方法:纳入COPD患者64例,其中男性51例、女性13例,COPD急性加重期患者组31例、稳定期患者组33例;另有健康对照25例.在入选时收集受试者的一般情况、测定肺功能、将诱导痰离心后留取痰液上清液测定C3a,对细胞沉渣进行炎症细胞分类计数.结果:COPD急性加重期组的诱导痰中C3a水平较COPD稳定期组及对照组升高,分别为44.2(13.2～48.5)μg/L、5.5(3.6～20.7)μg/L、1.7(0.1～5.9)μg/L,组间差异有统计学意义(Z=-2.974、-3.145,P=0.003、0.002).COPD稳定期组的诱导痰中C3a水平较对照组升高,但两组间差异无统计学意义(P＞0.017).健康对照组中的吸烟者与非吸烟者的诱导痰中C3a水平分别为1.97(0.12～6.27)μg/L、1.36(0.09～5.57)μg/L,两者差异无统计学意义(P＞0.05).COPD急性加重期患者组的诱导痰中C3a水平与炎症细胞总数呈正相关(r=0.543,P＜0.05).结论:C3a在COPD患者诱导痰中的水平增高,且与疾病的严重程度相关,提示其在疾病发生、发展中可能存在重要的作用.     

Detection of joint pathology by magnetic resonance imaging in patients with early rheumatoid arthritis

Magnetic resonance imaging (MRI) permits the visualization of anatomical structures not appreciated by conventional radiographic imaging, and may assess inflammatory disease and its progression with greater sensitivity than conventional radiography. In this study of 30 patients with early rheumatoid arthritis (RA), which could be considered as a pilot study because of the relatively small number of patients, we compare MRI of the knee and the fifth metatarsophalangeal joint with clinical and radiographic findings. A parallel study of 10 healthy individuals served as a reference group. In all but one of the 30 patients, MRI revealed some kind of joint abnormality, whereas conventional radiography was normal in 14 patients. The present study thus suggests that MRI may detect inflammatory and/or destructive joint changes in patients with early RA, and that these changes may occur in the absence of clinical symptoms or signs and/or radiographic signs in the examined joint. If these data prove to be confirmed in further controlled studies, MRI may be of importance both for the assessment of prognosis and for the decision to treat in the early critical stages of RA.     

Neutrophil collagenase in sputum from patients with cystic fibrosis

The potential role of neutrophil elastase in causing lung damage and exacerbating the inflammatory response in cystic fibrosis (CF) has received considerable attention. Although another potent neutrophil-derived enzyme, collagenase, is implicated in tissue destruction in several interstitial lung disorders, there has been no reference to this enzyme in CF. The objective of this study was to determine whether neutrophil collagenase is present in active form in CF sputum and, if so, whether it is related to disease severity. High levels of active collagenase were detected in sputum from patients with CF, and the majority of the enzyme present was of neutrophil origin. In a group of 16 patients with CF, negative relations between sputum collagenase activity and Shwachman score (r = -0.55, p < 0.05) and FEV1 (r = -0.59, p < 0.02) were noted, indicating an association between high collagenase activity and severity of disease. A positive correlation was observed between sputum collagenase and elastase activity (r = 0.62, p < 0.05). These results suggest that both neutrophil elastase and collagenase may play a significant role in lung destruction in CF.     

Serum levels of eosinophil cationic protein and eosinophil protein x in pollen atopic patients with stable asthma and its relation with bronchial hyperresponsiveness

Eosinophils are important effector cells in allergic inflammation described in allergic rhinitis (AR) and allergic bronchial asthma (BA). During the pollen season serum levels of eosinophil cationic protein (ECP) and eosinophil X protein/eosinophil-derived neurotoxin (EPX/EDN) are increased in BA. The aim of the present study was to evaluate the serum levels of ECP and EPC in pollen atopic patients with AR and BA during the winter. 92 patients were studied. They were divided into three groups: I 29 patients with AR, II 51 patients with BA and III 12 healthy subjects. Allergic rhinitis and bronchial asthma were diagnosed by routine clinical tests: clinical history, skin tests, total IgE and specific IgE. In addition ECP and EPX were determined in serum. All patients were asymptomatic, stable and without medical treatment. Methacholine challenge test (MCT) was performed in all patients. MCT were positive in 4 patients of group I and 45 patients of group II. ECP levels (ug/l) were: 21 (I), 24 (II) and 7 (III). EPX levels (ug/l) were 35 (I), 45 (II) and 21 (III). Statistical differences (p < 0.01) were observed both in ECP and EPX levels in patients with MCT positive in relation to patients with MCT negative, and in allergic patients (I and II) in comparison with the healthy subjects (III) (p < 0.01). ECP and EPX serum levels are increased in patients with a positive MCT in the winter, out of the pollen season, when patients are asymptomatic, stable and without treatment. This fact suggests that eosinophils play an important role in the pathogenesis of bronchial asthma.     

Stem cell factor-induced downregulation of c-kit in human lung mast cells and HMC-1 mast cells

Recent data suggest that local overexpression of the tissue-hormone c-kit ligand (stem cell factor [SCF]) is associated with accumulation of mast cells (MCs) and a decrease in expression of c-kit in the accumulated MCs [28]. In the present study, the effects of recombinant human (rh) SCF on expression of c-kit mRNA and c-kit protein in isolated human MCs and a human mast cell line, HMC-1, were analyzed. Incubation of isolated lung MC with rhSCF (100 ng/mL) for 120 minutes resulted in decreased expression of c-kit mRNA (optical density [OD], control: 100% vs. rhSCF: 37%). Almost identical results were obtained with HMC-1 cells (OD, control: 100% vs. rhSCF: 40 to 45%). As assessed by flow cytometry and monoclonal antibodies (mAbs) to c-kit, the SCF-induced decrease of c-kit mRNA in HMC-1 was associated with a substantial decrease in surface expression of c-kit (MFI, control: 100 +/- 21%, vs. MFI in cells incubated with rhSCF [100 ng/mL at 37 degrees C for 12 hours]: 8 +/- 2%, vs. MFI in cells incubated with rhSCF, 100 ng/mL, at 4 degrees C: 34 +/- 3%). The effects of rhSCF on c-kit expression in HMC-1 cells were dose- and time-dependent with maximum effects observed with 10-100 ng/mL of rhSCF after 4 to 12 hours. The SCF-dependent loss of c-kit was also accompanied by a decreased chemotactic response to rhSCF (control: 100%; rhSCF: 71 +/- 2%). This study shows that exposure of human lung MC and HMC-1 cells to recombinant SCF results in downregulation of c-kit mRNA and surface c-kit expression. These data may explain the partial loss of c-kit on MCs in areas of SCF overexpression.     

Lactic dehydrogenase virus infection enhances parasite egg production and inhibits eosinophil and mast cell responses in mice infected with the nematode Nippostrongylus brasiliensis

OBJECTIVE: To determine if awareness of methods to reduce vertical transmission of human immunodeficiency virus (HIV) is associated with HIV test acceptance and to clarify patients' attitudes toward routine versus elective prenatal HIV testing. METHODS: In a cross-sectional study, 247 antenatal patients were surveyed regarding HIV knowledge, self-perceived HIV risk, and willingness to learn a positive test result. This information, along with demographic and risk factor data, was related to HIV test acceptance. Patients also indicated their attitudes toward routine versus elective prenatal testing for HIV and other common prenatal screening tests. RESULTS: Seventy-two percent of antenatal patients accepted HIV testing. Test acceptance was not associated with the presence of risk factors, self-perceived HIV risk, or demographic factors, including race and ethnicity. Test acceptance was associated positively with patients' knowledge of a medical intervention to reduce vertical transmission and their willingness to learn a positive HIV test result. Only 24% of patients knew that the risk of vertical transmission could be reduced using medication. Sixty-nine percent of patients said that prenatal HIV testing should be routine, whereas 27% said that it should be done only after specific written consent. As a group, our patients viewed HIV screening no differently from screening for other infections in pregnancy. CONCLUSION: Interventions aimed at increasing HIV testing rates among pregnant women should focus on educating patients about vertical transmission reduction and promising new therapies for HIV infection. Proponents of elective testing should re-evaluate the assumption that patients view HIV testing differently from other prenatal tests for which separate written consent is not required.