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BACKGROUND: The diagnostic and staging approach for the mediastinal lymphadenopathies, with or whithout pulmonary lesions endoscopically visible, is based on transbronchial needle aspiration (TBNA) during fiberoptic bronchoscopy and on mediastinoscopy. One important factor impacting on TBNA sensitivity is the rapid on site cytological examination (ROSE). AIM: The aim of this study was to evaluate the economic impact of TBNA and TBNA + ROSE, in the diagnosis of these lesions. PATIENTS AND METHODS: 120 patients, affected by mediastinal lymphadenopathies suspected for lung cancer, underwent TBNA during fiberoptic bronchoscopy: 60 patients without ROSE (group A) and other 60 with ROSE (group B). Whenever needle aspirations failed to provide diagnosis, the patient underwent mediastinoscopy. The economic impact of the diagnostic process was performed. RESULTS: In group A, 39 patients (65%) obtained a diagnosis with TBNA while 21 patients (35%) required mediastinoscopy. In group B, 48 patients (80%) obtained a diagnosis with TBNA + ROSE, while 12 patients (20%) required mediastinoscopy. With regards to the costs of the procedures performed in the diagnostic process, the use of TBNA with ROSE as first diagnostic approach has saved a considerable amount of euros (19,413) compared to the use of TBNA without ROSE and the combined procedure increased (p < 0.02; chi square test) the sensitivity of TBNA by 15%. CONCLUSIONS: ROSE significantly impacts on the diagnostic yield, as well as on the overall management costs of patients with mediastinal lymphadenopathy, suspected for lung cancer. [ABSTRACT FROM AUTHOR]

Introduction: Lipoid pneumonia is a clinical condition that may be initially asymptomatic or confused with an infectious or malignant lung disease., Objectives: We report four cases of this pathological condition., Methods: The first case concerned an 85-year old woman with bilateral confluent pulmonary opacities, ground-glass type. Diagnosis was based on the cytology of the bronchoalveolar lavage (BAL) fluid followed by its ultrastructural examination. The second case was a 47-year-old man with an isolated pulmonary nodule, which was surgically removed; the diagnosis of lipoid pneumonia was formulated on the basis of the histological and electron microscopy examination. The third case concerned a 73-year-old woman, with bilateral hypodense areas at the bases of the lungs where FDG PET/CT scan showed an increased uptake. Diagnosis was formulated by BAL cytology and electron microscopy examination. The fourth case was a 69-year-old man, who performed a virtual colonoscopy for diverticulosis putting in evidence a round mass (3 cm in diameter) with two small peripheral nodules, located in the pulmonary left lower lobe. The histopathological examination of transthoracic biopsy confirmed a lipoid pneumonia., Results and Conclusion: In all four cases, it was put in evidence a prolonged use of a nasal decongestant containing mineral oils. In literature, the most cases described are characterized by a subclinical evolution and were presented as ground glass opacities which evolve, in the later phases, in an interstitial involvement or in a peripheral mass, simulating a lung tumour., (© 2016 John Wiley & Sons Ltd.)

Late breast implant seroma may be the presentation of a breast implant-associated anaplastic large cell lymphoma (BI-ALCL), which claims for a prompt recognition. However, BI-ALCL diagnosis on fine-needle aspiration (FNA) might be challenging for pathologists lacking experience with peri-implant breast effusions. Sixty-seven late breast implant seromas collected by FNA from 50 patients were evaluated by Papanicolaou smear stain and immunocytochemistry on cell blocks. A diagnostic algorithm based on the cellular composition, cell morphology and percentage of CD30+ cells was developed. Histological evaluation of the corresponding peri-prosthetic capsules was also performed. Most of the effusions (91% of the samples) were classified as reactive and 9% as BI-ALCL. In the BI-ALCL cases, medium-to-large atypical cells expressing CD30 represented more than 70% of the cellularity, whereas in in the reactive effusions CD30+ elements were extremely rare (<5%) and consisted of non-atypical elements. The reactive effusions were categorized into three patterns: i) acute infiltrate with prominent neutrophilic component (33% of the samples); ii) mixed infiltrate characterized by a variable number of neutrophils, lymphocytes and macrophages (30% of the samples); iii) chronic infiltrate composed predominantly of T lymphocytes or macrophages with only sporadic granulocytes (37% of the samples). The inflammatory cytological patterns were consistent with the histology of the corresponding capsules. Our results indicate that cytological analysis of late breast implant effusions, supported by the knowledge of the heterogeneous cytomorphological spectrum of late seromas, is a valuable approach for the early recognition of BI-ALCL.

Transbronchial needle aspiration (TBNA) is a bronchoscopic technique allowing the sampling of cytological/histological material from mediastinal lymph nodes. TBNA is routinely used only in few centers for the staging of lung cancer, and even less frequently for the diagnosis of mediastinal metastases from extrapulmonary tumors. We illustrate 5 cases of mediastinal metastases from extrapulmonary tumors observed at our center in order to emphasize the usefulness of cytology and TBNA in the diagnosis of these pathologies. The 5 cases illustrated were: seminoma, uterine cervical carcinoma, pleural mesothelioma, pancreatic carcinoma, pericardial mesothelioma. In these 5 cases, albeit not of lung cancer, the cytology on TBNA allowed the rapid formulation of the correct diagnosis; its main advantage is that it can be performed during a simple fiberbronchoscopy under local anesthesia with less risk and at a lower cost than a computed tomography-guided needle biopsy or mediastinoscopy.

Serous effusions are frequently a clinical manifestation of metastatic disease, with lung, breast and ovarian carcinoma and mesothelioma leading the list. The diagnosis of malignant effusion signifies disease progression and is associated with a worsening patient prognosis. The ability to grow in a dense exudative fluid suggests that the malignant cells are capable of acquiring nutrients, surviving and proliferating, despite the lack of a solid-phase scaffold. During proliferation, neoplastic cells release ligands and matrix metalloproteinases (MMPs) into their environment, which dissolve the extracellular matrix (ECM). Tissue inhibitors of metalloproteinase (TIMPs) are endogenous regulators of MMPs, the principal enzymes responsible for the degradation of ECM in metastasis, and reduce their proteolytic activity. TIMP-2 has demonstrated an association between high tumor tissue expression levels and poor prognosis. The purpose of this preliminary study is to investigate, by immunocytochemistry, TIMP-2 expression in non-neoplastic and metastatic adenocarcinoma pleural effusions. We selected 16 cases of reactive mesothelio, 7 of normal mesothelio, 14 of lung adenocarcinoma, 9 from the ovary, 4 from the gastrointestinal tract and 3 from the breast. In 23/30 cases (76%), we detected adenocarcinoma cells with strong TIMP-2 expression. Positive TIMP-2 expression was found in 2/7 cases (28%) of normal and 2/16 (12%) of reactive mesothelio. A statistical association was detected between TIMP-2 expression and metastatic adenocarcinoma cells compared to reactive and normal mesothelial cells (p<0.00003). The calculated sensitivities for TIMP-2 compared to CEA and Ber-EP4 were, respectively, 76.7, 80.0 and 93.3%, and the specificities 82.6, 95.7 and 87.0%. In conclusion, immunocytochemical detection of TIMP-2 could be considered an interesting marker in metastatic adenocarcinoma pleural effusions, and could possibly be used as a component of an antibody panel in diagnostic cytopathology.

The gene of mammalian acetylcholinesterase (AChE) generates multiple molecular forms, by alternative splicing of its transcripts and association of the tailed variant (AChET) with structural proteins. In the mammalian brain, the major AChE species consists of AChET tetramers anchored to the cell membrane of neurons by the PRiMA protein (Perrier et al., 2002). Stress and anticholinesterase inhibitors have been reported to induce rapid and long-lasting expression of the readthrough variant (AChER) in the mouse brain (Kaufer et al., 1998). In the readthrough transcript, there is no splicing after the last exon encoding the catalytic domain, so that the entire alternatively spliced 3' region is maintained. It encodes a C-terminal peptide with no specific interaction properties: COS cells transfected with AChER produce a soluble, nonamphiphilic monomeric form. We quantified AChER and total AChE expression in the mouse brain after an immobilization stress and after heat shock in neuroblastoma cells, and compared the observed effects with those induced by irreversible AChE inhibition (Perrier et al., 2005).

In vertebrates, the catalytic domain of acetylcholinesterase (AChE) may be associated with several C-terminal peptides generated by alternative splicing in the 3' region of transcripts. The "readthrough" (R) variant results from a lack of splicing after the last exon encoding the catalytic domain. Such a variant has been observed in Torpedo and in mammals; its C-terminal r peptide, also called "AChE Related Peptide" (ARP), is poorly conserved between rodents and humans. In rodents, it is significantly expressed in embryonic tissues and at a very low level in the brain of adult mice; it may be increased under various stress conditions, but remains very low. The "hydrophobic" (H) variant generates glycolipid (GPI)-anchored dimers, which are expressed in muscles of Torpedo, and in blood cells of mammals; H variants exist in Torpedo and in mammals, but apparently not in other vertebrate classes, suggesting that they were lost during evolution of early vertebrates and re-appeared independently in mammals. The "tailed" (T) variant exists in all vertebrate cholinesterases and their C-terminal t peptides are strongly conserved; in mammals, AChE(T) subunits represent the major type of acetylcholinesterase in cholinergic tissues. They produce a wide variety of oligomeric forms, ranging from monomers to heteromeric assemblies containing the anchoring proteins ColQ (collagen-tailed forms) and PRiMA (membrane-bound tetramers), which constitute the major functional enzyme species in mammalian muscles and brain, respectively. The oligomerization of AChE(T) subunits depends largely on the properties of their C-terminal t peptide. These peptides contain seven conserved aromatic residues, including three tryptophans, and are organized in an amphiphilic alpha helix in which these residues form a hydrophobic cluster. The presence of a cysteine is required for dimerization, while aromatic residues are necessary for tetramerization. In the collagen-tailed molecules, four t peptides form a coiled coil around a proline-rich motif (PRAD) located in the N-terminal region of ColQ. The t peptide also strongly influences the folding and cellular trafficking of AChE(T) subunits: the presence of hydrophobic residues induces partial misfolding leading to inactive protein, while aromatic residues, organized or not in an amphiphilic helix, induce intracellular degradation through the "Endoplasmic Reticulum Associated Degradation" (ERAD) pathway, rather than secretion. It has been proposed that the r and t C-terminal peptides, or fragments of these peptides, may exert independent, non cholinergic biological functions: this interesting possibility still needs to be documented, especially in view of their various degrees of evolutionary conservation.

Background and Aim of the Study: Pulmonary sarcoidosis is a chronic granulomatous disease characterized by macrophage and CD4+ T-cell accumulation at the site of inflammation. Analysis of the cytokine network has substantially improved knowledge on immunopathogenesis of sarcoidosis. We hypothesize that neurotrophins (NTs), nerve growth factor (NGF), brain derived neurotrophic factor (BDNF) and NT-3, besides their importance in immune system activities, participate in chronic inflammatory disorders and in repair processes., Methods: The expression of NTs and NT receptors was assessed in broncho alveolar lavage (BAL) macrophages, CD4 and CD8 T-cells, from 10 patients with pulmonary sarcoidosis, using molecular biology, Western blotting and immunocytochemistry., Results: Increased levels of NTs and of high affinity NT receptor (Trks) transcripts and proteins in BAL macrophages, CD4+ and CD8+ T-cells from pulmonary sarcoidosis patients were demonstrated in comparison with healthy controls. Contrarily to healthy controls, in pulmonary sarcoidosis the expression of NGF was increased in alveolar macrophages as well as NGF and BDNF in CD4+ and CD8+ T-cells. An increased expression of TrkA, TrkB and TrkC receptors was also noticeable. Furthermore, BDNF expression in alveolar macrophages and NT-3 expression in the three different BAL immune cell populations investigated were induced during sarcoidosis. A significant correlation was observed between CD4:CD8 ratio, lymphocytosis, radiological stage and CD4 and CD8 NT expression., Conclusions: These findings suggest that NTs are exaggeratedly expressed in BAL immune cells in pulmonary sarcoidosis and may participate in the progression of disease modulating immune cell functions.

Acetylcholinesterase (AChE) exists in various molecular forms, depending on alternative splicing of its transcripts and association with structural proteins. Tetramers of the 'tailed' variant (AChE(T)), which are anchored in the cell membrane of neurons by the PRiMA (Proline Rich Membrane Anchor) protein, constitute the main form of AChE in the mammalian brain. In the mouse brain, stress and anticholinesterase inhibitors have been reported to induce expression of the unspliced 'readthrough' variant (AChE(R)) mRNA which produces a monomeric form. To generalize this observation, we attempted to quantify AChE(R) and AChE(T) after organophosphate intoxication in the mouse brain and compared the observed effects with those of stress induced by swimming or immobilization; we also analyzed the effects of heat shock and AChE inhibition on neuroblastoma cells. Active AChE molecular forms were characterized by sedimentation and non-denaturing electrophoresis, and AChE transcripts were quantified by real-time PCR. We observed a moderate increase of the AChE(R) transcript in some cases, both in the mouse brain and in neuroblastoma cultures, but we did not detect any increase of the corresponding active enzyme.

Despite numerous studies demonstrating an association between asthma and many other chronic conditions and signs of Chlamydia pneumoniae (Cp) infection, the role of Cp in the pathogenesis of these illness remain still unclear. We investigated the prevalence of Cp antigen in the upper airways and the prevalence of detectable Cp serum antibodies in an unselected population of 207 9-yr-old schoolchildren. We also sought the presence of asthma, chronic or recurrent respiratory symptoms by means of questionnaire completed by the parents. Nasal aspirate, blood sampling and allergen skin prick tests were also performed. None of the children had obvious signs of acute infection at physical examination. Cp DNA was detected in nasal aspirates from 20 of the 207 children tested and serum IgG antibodies for Cp in 68 children. No association was found between atopy or history of atopic illness and the presence of Cp DNA or antibody production. This finding is explained by the fact that our study was conducted in an unselected childhood population, inherently including few children with asthma. A strong association between the status of antigen carrier and the presence of detectable Cp serum immunoglobulin (Ig)G or IgM suggests that subjects with detectable Cp antibodies have an impaired ability to eliminate this pathogen when infected. Because Cp eradication requires a strong Th1 lymphocyte response, the previously proven association between Cp and asthma, might reflect the known association of asthma with Th2-oriented lymphocytic activity., (Copyright (c) 2005 Blackwell Munksgaard)

The C-terminal 40-residue t peptide of acetylcholinesterase (AChE) forms an amphiphilic alpha helix with a cluster of seven aromatic residues. It allows oligomerization and induces a partial degradation of AChE subunits through the endoplasmic reticulum-associated degradation pathway. We show that the t peptide induces the misfolding of a fraction of AChE subunits, even when mutations disorganized the cluster of aromatic residues or when these residues were replaced by leucines, indicating that this effect is due to hydrophobic residues. Mutations in the aromatic-rich region affected the cellular fate of AChE in a similar manner, with or without mutations that prevented dimerization. Degradation was decreased and secretion was increased when aromatic residues were replaced by leucines, and the opposite occurred when the amphiphilic alpha helix was disorganized. The last two residues (Asp-Leu) somewhat resembled an endoplasmic reticulum retention signal and caused a partial retention but only in mutants possessing aromatic residues in their t peptide. Our results suggested that several "signals" in the catalytic domain and in the t peptide act cooperatively for AChE quality control.

Background: Previous studies have shown that histamine skin reactivity (the dimensions of a skin wheal elicited by a prick with histamine 10 mg/ml) in unselected school children has increased in Italy during the past two decades and is higher in Italy than in Poland. Hence this variable can probably be influenced by a changing or different lifestyle. The aim of this study was to compare skin reactivity to histamine and codeine (a marker of histamine releasability from mast cells) in schoolchildren from countries with different lifestyles., Methods: Six previously unstudied unselected populations of 9-year-old schoolchildren (two each from Poland, Italy, and Libya; n = 863 subjects; 49.0% males) were pricked with two concentrations of histamine (10 and 1 mg/ml) and codeine (90 and 9 mg/ml)., Results: The higher concentrations of both pharmacologic agents tested yielded significantly different wheal areas in the three countries: Poland < Italy < Libya (histamine, 11.8, 16.1 and 20.7 mm2; codeine, 9.2, 13.2 and 16.2 mm2; p < 0.001 for all comparisons). The lower concentrations elicited almost matching results. Histamine wheal areas correlated closely with areas elicited by codeine in the same individual: angular coefficients of the histamine to codeine regression lines were 0.535, Italy; 0.551, Libya; 0.612, Poland; and 0.581 for the whole population. More histamine was needed to produce a wheal in Poland than in Libya: a 20-mm2 wheal required an injected histamine concentration of about 8.8 mg/ml in Libya, 29.5 mg/ml in Italy and 102.1 mg/ml in Poland., Conclusion: More studies are necessary to explain the observed international differences in skin histamine reactivity and their effect on the prevalence of positive allergen skin tests.

The C-terminal t peptide (40 residues) of vertebrate acetylcholinesterase (AChE) T subunits possesses a series of seven conserved aromatic residues and forms an amphiphilic alpha-helix; it allows the formation of homo-oligomers (monomers, dimers and tetramers) and heteromeric associations with the anchoring proteins, ColQ and PRiMA, which contain a proline-rich motif (PRAD). We analyzed the influence of mutations in the t peptide of Torpedo AChE(T) on oligomerization and secretion. Charged residues influenced the distribution of homo-oligomers but had little effect on the heteromeric association with Q(N), a PRAD-containing N-terminal fragment of ColQ. The formation of homo-tetramers and Q(N)-linked tetramers required a central core of four aromatic residues and a peptide segment extending to residue 31; the last nine residues (32-40) were not necessary, although the formation of disulfide bonds by cysteine C37 stabilized T(4) and T(4)-Q(N) tetramers. The last two residues of the t peptide (EL) induced a partial intracellular retention; replacement of the C-terminal CAEL tetrapeptide by KDEL did not prevent tetramerization and heteromeric association with Q(N), indicating that these associations take place in the endoplasmic reticulum. Mutations that disorganize the alpha-helical structure of the t peptide were found to enhance degradation. Co-expression with Q(N) generally increased secretion, mostly as T(4)-Q(N) complexes, but reduced it for some mutants. Thus, mutations in this small, autonomous interaction domain bring information on the features that determine oligomeric associations of AChE(T) subunits and the choice between secretion and degradation.

Several studies have shown a higher prevalence of positive skin-prick tests to airborne allergens in Western than in Eastern European countries. We have recently reported that skin histamine reactivity significantly increased in Italy over the past 15 years. Population differences in skin histamine reactivity could, at least in part, explain the reported differences in positive allergen skin tests. To test this hypothesis we compared histamine skin reactivity and the prevalence of allergen positive skin-prick tests in a sample of Italian and Polish schoolchildren. A total of 336 unselected 9-year-old-schoolchildren (198 in Italy and 138 in Poland) underwent skin-prick tests with three different histamine concentrations (10, 1 and 0.2 mg/ml) and with a panel of common airborne allergens according to the ISAAC protocol, phase two. Mean wheals elicited by skin-prick tests with the three serial concentrations of histamine were significantly larger (p < 0.001) and shifted more toward higher values (p < 0.001) in Italian than in Polish children. The differences were greater for the intermediate histamine concentration tested (1 mg/ml) than for the highest concentration (10 mg/ml). Skin-prick tests for airborne allergens were more frequently positive in Italian children: wheals >or= 3 mm induced by any allergen [odds ratio (OR) 1.69; confidence interval (CI) 0.98-2.92] by Dermatophagoides pteronyssinus (OR 1.92; CI 0.97-3.80) and by D. farinae (OR 3.15; CI 1.16-8.63). Labeling as positive allergen wheal reactions half the size of the 10 mg/ml histamine wheal or larger reduced but did not abolish the Italian-Polish differences. The significantly higher skin histamine reactivity observed in Italian children could help to explain why allergen skin-test reactions differ in the East and West European populations. Moreover, differences in nonallergen-specific factors among populations should be considered in the interpretation of skin test results (e.g. cut-off points). To obtain meaningful results, epidemiological studies of allergies should include serial histamine dilutions.

In this paper we describe the gene organisation of sea bass Dicentrarchus labrax interleukin-1beta (IL-1beta) and the presence of incompletely spliced forms of this gene. Interestingly, the sea bass IL-1beta gene comprises five exons and four introns, thus being remarkably different from other known teleost and mammalian genes. The sizes of the introns were typically much shorter than in mammals and this feature, together with the loss of two introns, gave a much smaller gene than in mammals (2.7 vs 6.5-7.0kb). The 5'-end exon-intron organisation seems quite different in the known IL-1beta genes with three exons in human and carp, two in trout and only one in sea bass. The highest percentage of exon identity and similarity is between sea bass exon 4, trout exon 5, carp, mouse, chicken, pig and human exon 6. RT-PCR analysis revealed the presence of two incompletely spliced transcripts. Southern blot analysis suggests the presence of only one copy of IL-1beta gene in sea bass genome.

Objective: Although rhinitis is extremely frequent in children, methods for assessing the severity of nasal inflammation produce results with wide variability and hence weak clinical significance. We designed this epidemiologic investigation to define the clinical usefulness of assessing nasal cellularity in children., Methods: We studied 183 of 203 eligible unselected schoolchildren who were aged 9 to 11 years and whose parents gave informed consent and completed a questionnaire on the history of atopic and respiratory symptoms. In all children, nasal swabs were obtained from both nostrils and eluted in saline and slides were prepared from cytospin preparations for staining and white cell counts. Children also underwent determination of nasal volume, skin prick tests with 7 common local allergens, flow volume curves, and nitric oxide measurement in expired air. Blood samples were drawn for the measurement of total immunoglobulin E, eosinophil percentage, and detection of Chlamydia pneumoniae antibodies. C pneumoniae DNA was also sought in eluates from nasal swabs. The percentage, standard deviations, and percentiles of the various nasal white cell populations were determined., Results: No correlation of the percentage of these cells was found with the history of allergies or respiratory disease or with functional or laboratory finding. Repeat nasal swabs obtained 1 month after the initial examination in 31 children (20 with neutrophils higher and 11 lower than 14%) in 77.4% of the cases confirmed the previous (high or normal) result. Twelve of the 16 eligible children with persistently high nasal neutrophil counts completed a 15-day cycle of intranasal flunisolide therapy (200 micro g twice a day). Therapy significantly reduced nasal neutrophil percentage and increased nasal volume., Conclusions: Increased nasal neutrophils, although related neither to the clinical history nor to laboratory variables, are a common important finding in children. A 15-day cycle of intranasal flunisolide is sufficient to restore normal nasal neutrophilia.

Epidemiological information on symptoms affecting extra-respiratory organs and apparatuses in asthmatic children is scarce. The aim of this study therefore was to evaluate, at a population level, if and what extra-respiratory symptoms are associated with asthma. Two questionnaire-based, cross-sectional surveys were carried out on 1,262 students (651 males; mean age 9.57 years, age-range 6-14 years) in 1992 and on 1,210 students (639 males; mean age 9.02 years, age-range 6-14 years) in 1998, from two elementary and two junior high schools in Rome, Italy. Questionnaires included queries about asthma and its risk factors and extra-respiratory symptoms (headache, restlessness, sleep disturbances, urticaria, itching, and abdominal pain). Of responders, 11.9% (279/2,342) had a history of asthma. After adjustment for gender, family history of atopic disease, low birth weight, early respiratory problems, and damp house, asthma was significantly associated with recurrent abdominal pain (odds ratio [OR] 1.90; 95% confidence interval [CI]: 1.04, 3.16), itching (OR 3.15; 95% CI: 1.75, 5.68), and urticaria (OR 2.52; 95% CI: 1.02, 6.20). Asthma was reported by 10.2% (201/1,962) of children unaffected by this triad, by 20.1% (56/279; OR 2.20) with one of the symptoms, and by 31.6% (12/38; OR 4.04) with two or more symptoms. An emerging characteristic of pediatric asthma in our setting appears to be its association with certain extra-respiratory symptoms (abdominal pain, itching, and urticaria). A global, internistic approach to asthmatic children is increasingly required both in the clinical setting and in future epidemiological studies.

Asthmatic bronchial inflammation is associated with increased nitric oxide concentrations in exhaled air (eNO). Recent data suggest that this effect arises from atopy. Our aim in this study was to find out whether atopy and sensitization to particular allergens influences eNO levels. A total of 213 subjects (41 asthmatics and 172 controls) (96 boys and 117 girls, 7.3-14 years of age) were studied. Parents completed a questionnaire that sought information on their children's respiratory symptoms and exposure to tobacco smoke. Subjects underwent skin-prick tests for the following common allergens: Dermatophagoides pteronyssinus (Dpt), cat fur, Aspergillus fumigatus, Alternaria tenuis, mixed grass, mixed tree pollen, Parietaria officinalis, egg, and cow's milk. eNO was collected in 1-l mylar bags (exhaled pressure 10 cmH2O, flow 58 ml/s) and analyzed by using chemiluminescence. Atopic and non-atopic children without a history of chronic respiratory symptoms had a similar geometric mean eNO (atopics, n = 28, 11.2 p.p.b.; non-atopics, n = 96, 10.0 p.p.b.; mean ratio 1.1, 95% confidence interval [CI]: 0.7-1.6). Conversely, atopic asthmatic subjects had significantly higher eNO values than non-atopic asthmatic subjects (atopics, n = 25, 24.8 p.p.b.; non-atopics, n = 16, 11.4 p.p.b.; mean ratio 2.2, 95% CI: 1.2-3.9, p= 0.000). In children with rhinitis alone (n = 15) and those with lower respiratory symptoms other than asthma (n = 33), eNO increased slightly, but not significantly, with atopy. eNO levels correlated significantly with Dpt wheal size (r = 0.51) as well with the wheal size for cat, mixed grass, and Parietaria officinalis (r = 0.30-0.29), and with the sum of all wheals (r = 0.47) (p= 0.000). Subjects sensitized only for Dpt (but not those subjects sensitized only for grass pollen or other allergens) showed significantly higher eNO levels than non-atopic subjects (16.4 p.p.b. vs. 10.2 p.p.b., mean ratio 1.6, 95% CI: 1.1-2.3, p= 0.002). In asthmatic subjects, Dpt sensitization markedly increased eNO levels (Dpt-sensitized subjects: 28.0 p.p.b.; Dpt-unsensitized subjects: 12.2 p.p.b.; mean ratio 2.3, 95% CI: 1.5-3.5, p= 0.000). Non-asthmatic Dpt-sensitized subjects also had significantly higher eNO values than non-asthmatic, non-Dpt-sensitized subjects (14.2 p.p.b. vs. 10.1 p.p.b.; mean ratio 1.4, 95% CI: 1.1-1.9, p= 0.008). No difference was found between eNO levels in asthmatic subjects and control subjects exposed or unexposed to tobacco smoke. In conclusion, eNO concentrations are high in atopic asthmatic children and particularly high in atopic asthmatics who are sensitized to house-dust mite allergen.

The concentration of exhaled nitric oxide (eNO) is a useful marker of asthmatic bronchial inflammation. eNO can now be measured away from the laboratory (off-line), even in children. Short exhalation maneuvers (8 sec) and small samples (1 L) of exhaled gas are probably sufficient in children, but more information is needed about the effect of different measurement conditions. As a preliminary step before conducting epidemiological studies in schoolchildren, we investigated the effects of expiratory flow, dead space, and expiratory time on eNO concentrations collected in 1-L mylar collection bags. We studied 101 cooperative subjects (62 males) aged 5-18 years (30 healthy volunteers, 51 asthmatics, and 20 children with various other respiratory diseases) in our pulmonary function laboratory. On-line and off-line eNO were compared in a single session, and analyzed with a Sievers NOA 280 nitric oxide analyzer. For both methods of collecting expired gas, subjects did a single exhalation without breath-holding against an expiratory pressure 10 cm H(2)O. We investigated the effects of expiratory flow, dead space, and exhalation time on eNO; we also compared on-line and off-line eNO measurements, and the repeatability of both techniques at a given flow rate. Expiratory flows of 58 mL/sec provided more reproducible data than lower flows (coefficient of repeatability 1.1 ppb for 58 mL/sec vs. 2.8 for 27 mL/sec vs. 5.7 for 18 mL/sec). eNO concentrations were about 25% higher in off-line than in on-line recordings if the initial 250 mL of exhaled gas were not eliminated, and 37% higher if exhalation lasted longer (16 sec vs. 8 sec). Eliminating 250 mL of dead space and shortening the filling time to 8 sec yielded off-line eNO values close to those on-line (geometric mean off-line eNO 14.4 ppb, 95% confidence interval: 12.2-17.0) vs. on-line eNO 13.8 ppb (95% confidence interval: 11.6-16.5). On-line and off-line results were highly correlated (r = 0.996, P = 0.000) and had similar coefficients of variation (on-line eNO 2.6%, off-line 2.8%). Neither agreement nor repeatability of eNO measurements were affected by disease status or baseline FEV(1) (% predicted values). Once standardized, the off-line eNO technique using 1-L gas collection bags will provide results similar to those recorded on-line., (Copyright 2001 Wiley-Liss, Inc.)

Time trends in the prevalence of asthma, family history of asthma and atopy in Roman schoolchildren were assessed. The study population consisted of all children (aged 6-14 yrs) attending two primary schools in Rome, situated in urban areas that differed markedly in socioeconomic conditions and environmental pollution. Three questionnaire-based surveys were conducted in 1974, 1992 and 1998 in 2,259, 1,229 and 1,139 children. The prevalence of asthma in males and females increased significantly during 1974-1992 and remained stable from 1992-1998. In age groups born in the subsequent 4-yr periods it increased almost linearly, for children born from 1962-1965 to 1982-1985 (4.4%-12.5%), and remained remarkably stable in children born after 1985. Because the prevalence of asthma had a steeper trend in males than in females (approximately 0.55% x yr(-1) versus 0.25% x yr(-1)), the male:female asthma ratio increased (1:38 in 1974; 1:84 in 1992 and 1:62 in 1998). No single environmental factor, including area of residence, seemed to influence the prevalence of asthma. Family history of asthma and atopy also increased steadily (0.72% x yr(-1) and 0.30% x yr(-1) respectively) more than doubling during the 24-yr study period. The strong relationship between asthma and a family history of atopy not only persisted but also strengthened over time (23.3% of asthmatic children belonged to families with atopic illnesses in 1974 but 44.2% in 1998). The environmental factors that might explain the almost three-fold rise in childhood asthma between 1974 and 1992 remain unknown but the genetic background of the disease has presumably remained unchanged since the early 1970s. The fact that the prevalence of asthma increased no further during the past 6 yrs suggests that the progressive induction of asthma symptoms in genetically predisposed subjects is a self-limiting process that has probably come to an end in the authors' study area.

Children with acquired immune deficiency syndrome (AIDS) commonly have recurrent infectious and noninfectious lung complications that ultimately end in death. To study the intensity of alveolar inflammation and to evaluate the clinical utility of bronchoalveolar lavage (BAL) in children with HIV-1 infections, we retrospectively analyzed differential cell counts, lymphocyte subsets, and fibronectin and hyaluronic acid concentrations in BAL fluid of 18 HIV-1-positive children (9 boys, mean age 3.5 years, range 5 months-8 years) with radiological evidence of interstitial lung disease, and 19 control children who had undergone BAL for clinical indications not involving the lung parenchyma (13 boys, mean age 3 years, range 2 months-14 years). BAL fluid from 89% of the HIV-1 infected children showed CD8+ve lymphocytic alveolitis expressing HLA-DR, CD54, and CD 69 antigens. BAL fluid from HIV-infected patients typically contained markedly increased percentages and numbers of lymphocytes (P < 0.0001) and eosinophils (P < 0.04) and significantly higher concentrations of albumin (P < 0.05) and fibronectin (P < 0.0006) than fluids from control children. Whereas BAL cellular components did not differ in P. carinii-positive and P. carinii-negative HIV-1-infected children, fibronectin concentrations were significantly higher in P. carinii-positive than negative children. BAL cell differentials and noncellular components were related neither to severity of disease nor to patients' disease progression. These findings indicate that BAL is useful in studying the intensity of lung inflammation in children with HIV-1 infections and radiologically documented interstitial lung disease, but provides no information on the subsequent clinical course., (Copyright 2001 Wiley-Liss, Inc.)

In an asymptomatic 4 yr old child with radiographic evidence of parenchymal lung disease, bronchoalveolar lavage (BAL) yielded the diagnosis of chronic lipid pneumonia caused by chronic aspiration of mineral oil given as a laxative. BAL analysis showed a marked reduction in the total number of alveolar macrophages; almost 70% of these cells contained intracytoplasmic lipid vacuoles. It also disclosed lymphocytic (cytotoxic/suppressor) alveolitis. A high percentage of lymphocytes expressed antigen markers of activation (human leucocyte antigen (HLA)-DR), CD54 and CD25). BAL analysis 18 months after mineral oil intake revealed that lymphocytes bearing antigen markers of activation had markedly decreased whereas alveolar macrophages (normal and lipid-laden) had increased. A subsequent whole lung BAL was considered unnecessarily invasive in this otherwise healthy child.

The american primate Cebus apella has been used as an experimental model for the study of acute and chronic Chagas' disease. The antibody response elicited by 4 x 10(6) blood trypnomastigotes injected into four monkeys was analysed. Peak titres of IgM and IgG of anti-Trypanosoma cruzi antibodies were found at day 22, and between days 20 and 40 post-infection (p.i.), respectively. The ability of a Mr 37kDa (T37K) glycoprotein purified from T. cruzi epimastigotes to generate IgG anti-T. cruzi antibodies in monkeys, and protect them against a challenge with trypomastigotes, was also studied. Monkeys non-immunized with T37K reached peak values of parasitaemia between days 18 and 21 post-infection, whereas immunized monkeys had lower parasitaemias without important variation. Anti-T37K antibodies in immunized monkeys decreased from day 2 with the lowest titres between days 14 to 22 p.i., coincident with the peak of parasitaemia in control non-immunized monkeys. These results suggest that anti-T37K antibodies could be responsible for the low parasitaemia detected in immunized monkeys.

The objective was to study the secretory pattern, both basal and stimulated either by histamine (0.1 mg/kg) or pentagastrin (64 micrograms/kg) in eighteen Cebus apella monkeys chronically infected with different T. cruzi strains (CA1, n = 10; Colombian, n = 4 and Tulahuen, n = 4) and to describe the morphological findings in the gastrointestinal tract in twelve infected (6 sacrificed and 6 spontaneously dead) and four healthy monkeys. All infected monkeys and 35 healthy ones were evaluated by contrast X-ray examination. No differences were observed in basal acid output between control and infected groups. Animals infected with the Tulahuen and Colombian strains showed significant lower values of peak acid output in response to histamine or pentagastrin (p < 0.01 and p < 0.05 respectively; "t" test) in comparison to the controls. Barium contrast studies showed enlargement and dilatation of the colon in three infected animals. Histopathological lesions were seen in 75% of the autopsied animals either in colon alone (33%) or both, in colon and esophagus (42%). The normal secretion observed in the CA1 infected group could be due to a lower virulence of the strain, a lower esophageal tropism or the necessity of a longer post-infection time to cause lesions.

Purpose: To evaluate electrocardiographic data of Cebus apella monkeys with Chagas' disease., Material and Methods: 53 Cebus apella monkeys (juvenile and adult of both sexes) were used: 35 as a control group and 18 inoculated four-five years ago with 3 different Trypanosoma cruzi strains (CA1, n = 10; Colombian, n = 4; Tulahuen, n = 4)., Results: The normal electrocardiogram (ECG) showed differences with that of man, a) high cardiac rate; b) presence of "pulmonary p" wave without pulmonary pathology. The ECT alterations found between 11 and 58 months after last inoculation were: right bundle branch block; intermittent right bundle branch block; left ventricle overload; repolarization disturbances; left anterior hemiblock; extra systole. These alterations resemble those found in humans, as well as clinical parasitological and immunological alterations. Their incidence and the time at which they appeared, seem to vary according to the route, strain, inoculum and frequency of the inoculation. Three of the monkeys died spontaneously 46, 48 and 52 months after the infections due to the natural evolution of the disease, and six were sacrificed during the follow-up. In both cases histopathological lesions were found, and their intensity was directly related to the time and resembled the human disease., Conclusion: The Cebus apella, as it reproduces human electrocardiographic and histopathological alterations, a short time after experimental infection, is a suitable model for the study of the different aspects of the physiopathology, immunopathology and therapeutics of the indeterminate and chronic phases of Chagas' disease.

Eighteen Cebus apella monkeys, (juvenile and adult of both sexes) were inoculated five years ago, with three Trypanosoma cruzi strains (CA1, n = 10; Colombian, n = 4 and Tulahuen, n = 4), either by conjunctival or intraperitoneal route, once or repeatedly. Parasitological, hematological, serological, enzymatic, radiographic, electro and echocardiographic findings have been previously published and they are similar to those observed in human pathology. The most frequent electrocardiographic alteration was right branch bundle block. Six animals, chosen at random, were sacrificed. Those sacrificed 20 to 25 months post-first inoculation showed focal accumuli of leukocytes with myocytolysis. Foci of diffuse interstitial fibrosis with mild infiltrate of leukocytes among fibers were observed in the animals sacrificed 36 to 47 months post-inoculation. No parasites were seen. The lesions were more prominent in the ventricular walls and the septum. The fact that the infiltrates were predominant in the animals sacrificed at a shorter time after first inoculation and that fibrosis was more severe in those sacrificed at a longer time suggests that there is a progression of the infiltrative lesions to fibrosis, with a leukocytic activity indicative of a chronic phase. These lesions are similar to those described in human chronic Chagas' disease. This would demonstrate that this model is useful in evaluating a progress in the knowledge of the pathogenesis which is still a controversial issue, immunology, immunogenesis and chemotherapeutic agents of the chronic and indeterminate phases of this disease.

The aims of the present study were to determine in 14 healthy subjects and 14 duodenal ulcer patients the reproducibility of the acid secretory response to a modified sham-feeding test and the effect on this response of intrajejunal hypertonic glucose instillation, in order to evaluate the possibility of the existence of a defective inhibition of the cephalic phase of gastric acid secretion in duodenal ulcer disease. The reproducibility of the acid secretory response to a modified sham-feeding test was demonstrated in both groups in two consecutive tests. The hypertonic glucose instillation produced a significant inhibition of the acid secretory response to modified sham feeding only in the healthy subjects, suggesting that duodenal ulcer patients may have a defective mechanism of acid inhibition during vagal stimulation by modified sham feeding. Non-significant changes were observed in plasma gastrin and pancreatic glucagon levels.

Acid instillation into the duodenum inhibits basal and stimulated gastric secretion. In man vagotomy suppresses this secretory inhibition. It is postulated that such inhibition responds to a dual mechanism: an hormonal one (enterogastrone) and a nervous one (vagus nerve). This study showed that preoperative duodenal acidification of duodenal ulcer patients results in a decrease in basal gastric secretion and in gastrin levels. On the contrary, in patients submitted to vagal denervation--either through truncal division or highly selective vagotomy--duodenal acidification does not inhibit gastric secretion; however, a drop occurs in basal gastrin levels. An intact gastric vagal innervation therefore, seems necessary for the preservation of the sensitivity of the parietal cell to the effect of hormonal inhibitors, it being immaterial whether duodenal innervation is present or not, as that duodenal acidification provokes a significant fall in serum gastrin levels as determined by radioimmunoassay. This hormonal decrease produced by duodenal acidification can be explained by the inhibition of gastrin release from the antrum. Agreement is expressed with the opinion of other authors that highly selective vagotomy does not appear to carry any advantage over truncal section of the vagus nerves from the standpoint of the inhibitory mechanism of gastric secretion from the duodenum.

The humoral and cellular immunological parameters of the New World non-human primate Cebus apella were analysed. The study included: serum protein immunoelectrophoretic analysis; cross reactivity between monkey and human immunoglobulins by immunoprecipitation, ELISA and indirect immunofluorescence tests; immunoglobulin quantitation by radial immunodiffusion; and assays with peripheral blood lymphocytes involving tests for E and EAC rosettes and detection of surface markers (surface immunoglobulins and CD4-CD8 antigens). The results obtained showed that (a) at least three immunoglobulins with electrophoretic mobility corresponding to IgG, IgA and IgM which showed cross reactivity with the human ones were present in serum; (b) it was possible to evaluate the relative monkey immunoglobulin concentration using specific antibodies against human immunoglobulins and to obtain absolute values using adequate conversion factors; (c) lymphocytes forming E and EAC rosettes were found in peripheral blood in a similar proportion to that reported in man; (d) lymphocyte surface immunoglobulins were detected using anti-human immunoglobulin serum; (e) it was not possible to demonstrate the presence of T helper and T suppressor/cytotoxic lymphocytes using OK T4 and OK T8 monoclonal antibodies.

Recent reports cite an increased incidence in delayed gastric emptying following Roux-en-Y biliary diversion for alkaline reflux gastritis. The effect of Roux-en-Y diversion on the gastric emptying of liquids and solids was evaluated following vagotomy and antrectomy and vagotomy and subtotal gastrectomy. Twenty dogs underwent placements of large Thomas cannula in the stomach. Four dogs with intact stomachs served a controls. Eight dogs each with vagotomy and antrectomy were subdivided into Roux-en-Y gastrojejunostomy (RYA) and a Billroth II (B-IIA) group. Eight dogs each with vagotomy and subtotal gastrectomy were subdivided into similar groups. Four dogs - Roux-en-Y (RSTG) and four dogs - Billroth II (B-IISTG). Gastric emptying of solid food, normal saline and 25% dextrose was evaluated. RYA dogs demonstrated a significant delay in gastric emptying of solids compared with corresponding B-IIA animals. RYA dogs had 76, 61 and 42% of solid food retained at three, five and eight hours while B-II animals retained 56, 41 and 20%, respectively. The results are highly significant at all time intervals (p less than 0.001 at five and eight hours). Control animals retained 34, 17 and 3% of solid food at three, five and eight hours. RSTG animals had 73, 52 and 28% retained solids at three, five and eight hours, while B-IISTG animals had 55, 42 and 13% retention, respectively (p less than 0.05 at eight hours). Normal saline was significantly delayed in both Roux-en-Y subgroups compared with B-II dogs (p less than 0.02 in V/A, p less than 0.05 in V/STG). There was a trend toward delayed emptying of 25% dextrose in the Roux-en-Y groups, but significance was achieved only in the RYA compared with B-IIA groups (p less than 0.02 at 30 minutes). Delayed gastric emptying following Roux-en-Y gastrojejunostomy is documented in the experimental animals which underwent vagotomy and appears greater in magnitude than that observed following vagotomy and B-II gastrectomy. These data corroborate the clinical observations of severe delayed gastric emptying in patients undergoing Roux-en-Y diversions for alkaline gastritis.