Natural stranding of Atlantic sturgeon (Acipenser oxyrinchus Mitchill, 1815) in Scot's Bay, Bay of Fundy, Nova Scotia, from populations of concern in the United States and Canada.

Natural mortality of Atlantic sturgeon (Acipenser oxyrinchus) has been determined to be low (M = 0.07). Reported herein is the mortality by beach stranding of 11 Atlantic sturgeon in Scot's Bay, part of the inner Bay of Fundy in Nova Scotia, Canada on 22 June 2014. Genetic analyses, histological analysis and age determination were performed to determine origin, maturity stage and age of the stranded Atlantic sturgeon. Microsatellite and mitochondrial DNA analyses indicated that four of the Atlantic sturgeon (2 males and 2 females) were from the Saint John River, NB population, which was designated as threatened by the Committee on the Status of Endangered Wildlife in Canada. Seven Atlantic sturgeon (1 male, 5 females, 1 unknown) were from the Kennebec River, Maine population, that was listed as threatened under the Endangered Species Act in the U. S. Ageing of A. oxyrinchus by pectoral fin spine analysis determined that the mean age of the individuals from the Saint John River ( [Formula: see text] years, sd = 5.0) and the Kennebec River ( [Formula: see text] years, sd = 3.5) were not significantly different. This is the first report of a stranding event of Atlantic sturgeon, and describes a source of natural mortality affecting populations of concern in both Canada and the U. S.

Occupational hypersensitivity pneumonitis: an EAACI position paper.

The aim of this document was to provide a critical review of the current knowledge on hypersensitivity pneumonitis caused by the occupational environment and to propose practical guidance for the diagnosis and management of this condition. Occupational hypersensitivity pneumonitis (OHP) is an immunologic lung disease resulting from lymphocytic and frequently granulomatous inflammation of the peripheral airways, alveoli, and surrounding interstitial tissue which develops as the result of a non-IgE-mediated allergic reaction to a variety of organic materials or low molecular weight agents that are present in the workplace. The offending agents can be classified into six broad categories that include bacteria, fungi, animal proteins, plant proteins, low molecular weight chemicals, and metals. The diagnosis of OHP requires a multidisciplinary approach and relies on a combination of diagnostic tests to ascertain the work relatedness of the disease. Both the clinical and the occupational history are keys to the diagnosis and often will lead to the initial suspicion. Diagnostic criteria adapted to OHP are proposed. The cornerstone of treatment is early removal from exposure to the eliciting antigen, although the disease may show an adverse outcome even after avoidance of exposure to the causal agent.

Impaired function of regulatory T-cells in hypersensitivity pneumonitis.

Hypersensitivity pneumonitis (HP) is characterised by lung lymphocytosis. Most individuals exposed to HP antigens remain asymptomatic. The mechanisms involved in the impaired immune tolerance leading to HP are unclear. Normally, T-regulatory (Treg)-cells control the immune response. The aim of the present study was to determine whether Treg-cell suppressive function deficiency can explain the uncontrolled inflammation in HP. Bronchoalveolar lavage (BAL) and blood samples were obtained from normal subjects, asymptomatic individuals and HP patients. BAL and blood Treg-cells were isolated. The ability of Treg-cells to suppress T-cell proliferation and the role of interleukin (IL)-17 was verified. BAL and blood Treg-cells from normal subjects suppressed the proliferative response of activated T-cells by 47.1 and 42%, respectively. BAL and blood Treg-cells from asymptomatic subjects had a slightly decreased activity and suppressed proliferation by 29.4 and 31.8%, respectively. BAL and blood Treg-cells from HP patients were totally nonfunctional and unable to suppress proliferation. Low levels of IL-17 were detected in sera and BAL from both normal and asymptomatic individuals, whereas measurable levels were found in patients. Treg-cells may be involved in antigen tolerance in asymptomatic subjects. Defective Treg-cell function, potentially caused by increased IL-17 production, could account for the exacerbated immune response characteristic of HP.

Mature CD11c(+) cells are enhanced in hypersensitivity pneumonitis.

The present study verified the hypothesis that enhanced maturation of antigen-presenting CD11c(+) cells could explain the viral-induced exacerbated immune response to Saccharopolyspora rectivirgula (SR), the main antigen responsible for farmer's lung, a classic form of hypersensitivity pneumonitis (HP). Four groups of mice were studied: group 1 received intranasal instillations of saline; group 2 received instillations of SR for 12 weeks; group 3 received instillations of saline and a single infection with Sendai virus on week 3; and group 4 received instillations of SR for 12 weeks with a single administration of Sendai virus on week 3. On week 13, mice were sacrificed and bronchoalveolar lavage was performed. Lungs were harvested, digested with enzymes, and CD11c(+) cells were analysed in flow cytometry with anti-CD11c, anti-CD86 and anti-major histocompatibility complex class II markers. Immunofluorescence studies were also performed with the same cell surface markers. Both flow cytometry and immunofluorescence results demonstrate that mature CD11c(+) cells are significantly enhanced in SR-challenged mice simultaneously infected with Sendai virus, compared with other groups. These CD11c(+) cells persist in the lung for 9 weeks after the virus infection. Maturation of CD11c(+) cells could explain, at least in part, the virus-induced increased immune response to SR antigens in this model of HP, but mechanisms have still to be elucidated.

Hypersensitivity pneumonitis.

The first few cases of hypersensitivity pneumonitis (HP) were described in the early 20th century in farmers exposed to moldy hay or straw. As then, HP has been ascribed to multiple inhaled antigens found in a large variety of environmental settings. Hypersensitivity pneumonitis results from an exaggerated immune response, which gives rise to acute infection-like symptoms or to progressive, sometimes irreversible lung damage. The diagnosis is based on a combination of clinical characteristics of the disease. Clinical diagnostic criteria have recently been published. The immune mechanisms leading to HP are still incompletely understood. Initially, believed to be a classes III and IV immune response, we now have a clearer understanding of the complex inflammatory events involved. These include the release of pro inflammatory cytokines and a decrease in the immune control mechanisms via surfactant, dendritic and T-regulatory cells. Despite the improved understanding, the treatment and outcome of HP have not changed. Oral corticosteroids remain the only effective drugs and contact withdrawal constitutes the ideal solution. If unchecked, HP can lead to irreversible lung damage in the form of fibrosis or emphysema, respiratory insufficiency and even death.

Respiratory response to endotoxin and dust predicts evidence of inflammatory response in volunteers in a swine barn.

BACKGROUND: It has been shown that exposure in intense exposure in swine barn facilities is associated with increased respiratory symptoms and reduction in pulmonary functions. This study investigated if systemic response could be predicted by FEV(1) response following swine barn exposure. METHODS: Naïve males were tested at baseline, low and high endotoxin and dust levels. Subjects were classified as "more responsive" (n = 9) or "less responsive" (n = 11) based on FEV(1) reduction following high endotoxin exposure. Health measures included pulmonary function testing, blood samples and nasal lavage. Environmental samples were collected from the barn. RESULTS: White blood cells and blood lymphocytes at low exposure were significantly greater in those who were "more responsive" compared to those who were "less responsive". There was a significant increase in blood lymphocytes, serum IL6, total nasal lavage cells and nasal IL8 at high exposure among "more responsive" subjects compared to "less responsive" subjects. CONCLUSIONS: Respiratory response to high-level endotoxin and dust exposure predicts evidence of inflammatory response throughout a range of endotoxin and dust exposures.

Modulation of airway inflammation and resistance in mice by a nicotinic receptor agonist.

Nicotinic agonists, including 1,1-dimethyl-4-phenylpiperazinium (DMPP), have anti-inflammatory properties and in some instances smooth muscle relaxing effects. Since inflammation and airway smooth muscle contraction are two major components of asthma, the present authors investigated the effects of DMPP on airway inflammation and airway resistance in a mouse model of asthma. Mice were sensitised and challenged with ovalbumin (OVA) and treated either intraperitoneally or intranasally with DMPP. The effect of DMPP was tested on airway inflammation, airway resistance and on the increase of intracellular calcium in bronchial smooth muscle cells. DMPP given either during sensitisation, OVA challenges or throughout the protocol prevented lung inflammation and decreased the serum level of OVA specific immunoglobulin E. DMPP administration reduced the number of total cells, lymphocytes and eosinophils in the bronchoalveolar lavage (BAL) fluid. Intranasal DMPP administration was as effective as dexamethasone (DEXA) in reducing total cell count and eosinophil counts in BAL fluid. DMPP, but not DEXA, reduced tissue inflammation. Intranasal DMPP, given 10 min before the test, reduced airway responsiveness to metacholine. DMPP also reduced the increase in intracellular calcium in response to bradykinin. In conclusion, these results show that 1,1-dimethyl-4-phenylpiperazinium reduces lung inflammation and prevents airway hyperresponsiveness in the mouse model of asthma.

Effect of 1,1-dimethylphenyl 1,4-piperazinium on mouse tracheal smooth muscle responsiveness.

Bronchial hyperresponsiveness is one of the main features of asthma. A nicotinic receptor agonist, 1,1-dimethylphenyl 1,4-piperazinium (DMPP), has been shown to have an inhibitory effect on airway response to methacholine in an in vivo model of asthma. The aims of this study were to 1) verify whether nicotinic acetylcholine receptors (nAChR) were present on mouse tracheal smooth muscle, 2) verify whether bronchoprotection observed in mice was due to a direct effect on airway smooth muscle, and 3) compare the effects of nicotinic agonists to that of salbutamol. Alpha3-, alpha4-, and alpha7-nAChR subunits were detected by immunofluorescence on tracheal tissues from normal BALB/c mice. The effect of DMPP on tracheal responsiveness was verified by an isometric method. Tracheas were isolated from normal mice, placed in organ baths, and contracted with a single dose of methacholine. Cumulative doses of DMPP or salbutamol were added to the baths. Results show that mouse tracheal smooth muscle is positive for alpha4- and alpha7-nAChR subunits and that the epithelium is positive for alpha3-, alpha4-, and alpha7-subunits. DMPP induced a greater dose-dependent relaxation of tracheal smooth muscles precontracted with methacholine than with salbutamol. These results suggest that the smooth muscle-relaxing effect of DMPP could have some interest in the treatment of obstructive pulmonary diseases.

[Clinical and immunopathological aspects of hypersensitivity pneumonitis]. / Aspects cliniques et immunopathologiques des pneumopathies d'hypersensibilité.

INTRODUCTION: Hypersensitivity pneumonitis (HP) is a pulmonary disease with symptoms of dyspnoea and cough resulting from the inhalation of an antigen to which the patient has been previously sensitized. STATE OF ART: Acute and subacute HP represent the most active forms of the disease which may become chronic while remaining progressive. HP may also evolve to end-stage lung disease. Clinical symptoms and signs tend to be non-specific and the diagnosis of HP often relies on the clinical context. The immune response is initiated when the alveolar macrophage phagocytoses the antigen, provoking the expansion of lymphocytes T and B that reach the pulmonary parenchyma through the systemic circulation. This reaction is amplified by the expression of a number of inflammatory mediators. PERSPECTIVE AND CONCLUSION: This article summarizes our current understanding of the diagnostic approach and immunological mechanisms related to HP.

Occupational asthma in newly employed workers in intensive swine confinement facilities.

Respiratory symptoms, reductions in pulmonary function and increased bronchial responsiveness have been described in exposed workers and in naïve volunteers exposed to intensive swine production facilities. Typically, this occurs in persons who have been employed for a long duration or in previously unexposed, naïve volunteers. The current authors describe four cases, all female, who developed acute onset of wheezing and cough suggestive of asthma within weeks of commencing full-time employment in intensive swine production facilities. None of the workers were aware of any previous asthma, allergies or hay fever. All four employees reported improvement of symptoms on cessation of work in the facilities and consequent withdrawal from exposure. However, when seen at the respiratory clinic, cases 1 and 3 continued to be either mildly symptomatic or were taking medications with continued borderline airways responsiveness, as measured by methacholine challenge test up to 4 and 5 months, respectively, following work cessation. Case 2 continued to have symptoms for > or =3 months after work cessation. Only case 1, however, was seen at repeated visits in the respiratory clinic. One worker participated in a work re-entry trial and experienced profound coughing and chest tightness within an hour of entry, after which, the trial had to be terminated. Provocative concentration causing a 20% fall in forced expiratory volume in one second (PC20) measured 5 h later was lower than pre-trial PC20. No acute exposure event was recorded in the workers prior to the onset of symptoms. To the current authors' knowledge, this is the first report of occupational asthma occurring in newly employed full-time intensive swine production workers after a short-term exposure and should raise awareness that previously unexposed workers may be at risk of developing what would appear to be long-term asthma after relatively short-term exposure.

Significance of precipitins and asymptomatic lymphocytic alveolitis: a 20-yr follow-up.

What is the significance of serum precipitins and lymphocytic alveolitis in a healthy subject exposed to antigens responsible for hypersensitivity pneumonitis (HP)? This study was done to evaluate the 20-yr outcome of asymptomatic dairy farmers with or without precipitins or lymphocytic alveolitis in their bronchoalveolar lavage (BAL). Twenty-seven of the initial 43 farmers were restudied. Of the missing 16, 11 refused the follow-up, three had died and two could not be located. The restudied farmers had a clinical evaluation, lung function tests and a high resolution computed tomography (HRCT). Only one subject, an asthmatic, had dyspnoea, none described present or past symptoms suggestive of HP. Of those who were no longer on the farm, none had quit because of respiratory problems. Lung function changes were as expected with aging. The only difference was that farmers with positive precipitins had a statistically significant greater decrease in their forced expiratory volume in one second than the sero-negative farmers. Four HRCTs showed signs of minimal parenchymal abnormalities; these were not more prevalent for subjects with or without precipitins or BAL lymphocytosis. Serum precipitins and asymptomatic lymphocytic alveolitis in an asymptomatic, exposed dairy farmer have no clinically meaningful long-term consequences.

Apoptosis of bronchoalveolar lavage lymphocytes in hypersensitivity pneumonitis.

The aim of this study was to look at the apoptosis of alveolar lymphocytes in hypersensitivity pneumonitis (HP). HP patients and normal unexposed controls were studied. The percentage of apoptotic lymphocytes was significantly lower in HP patients than in normal patients (37.4 +/- 3.4 versus 56.5 +/- 5.5% for Annexin V and propidium iodine detection methods and 0.4 +/- 0.1 versus 1.0 +/- 0.2% for dUTP nick end-labelling technique (TUNEL)). The proportion of bronchoalveolar lavage (BAL) lymphocytes positive for Fas antigen was significantly higher in HP patients than in normal subjects (71.7 +/- 5.4 versus 50.4 +/- 9.0%). However, no significant difference was found in the proportion of BAL lymphocytes positive for Fas ligand (FasL) between the two groups. Soluble Fas (sFas) levels in the BAL fluid of the patients and normals were 80.5 +/- 8.5 pg x mL(-1) and 23.2 +/- 3.1 pg x mL(-1), respectively. A positive correlation was found between the percentage of BAL lymphocytes and the levels of sFas for the total subjects but not within the separate study groups. The intracellular quantity of the inducible anti-apoptotic gene Bcl-xL product was significantly higher in the pulmonary lymphocytes of HP patients than in lymphocytes of the control, while no difference was found for constitutive anti-apoptotic protein (Bcl-2). In conclusion, the apoptosis of pulmonary lymphocytes is lower in hypersensitivity pneumonitis than in normal subjects. This could be explained, at least in part, by an increase of soluble Fas, the anti-apoptic gene, and Bcl-xL.

Adaptation to organic dust exposure: a potential role of L-selectin shedding?

Swine confinement workers, exposed to high levels of organic dust, present a high prevalence of respiratory symptoms but show only mild lung inflammation. This contrasts with the intense inflammatory response observed when naive subjects are exposed to the same environment. Shedding of L-selectin may regulate the recruitment of inflammatory cells and explain this discrepancy. Soluble L-selectin (sL-selectin) levels were measured in sera of 36 workers, 35 control subjects and eight healthy volunteers briefly and repeatedly exposed to swine confinement buildings. White blood cell counts (WBC) and serum interleukin (IL)-6 levels were measured as markers of systemic inflammation. Higher concentrations of sL-selectin were found in the sera of workers than in controls (1452+/-62 ng x mL(-1) and 872+/-25 ng x mL(-1), respectively) whereas no differences were detected before and after acute repeated exposures of exposed volunteers. WBC were increased after exposure in exposed volunteers but not in workers. Both workers and exposed volunteers had increased IL-6 serum levels, although it was more pronounced for the exposed volunteers. These results support the hypothesis that shedding of L-selectin may downregulate the inflammatory response to organic dust-contaminated environments and constitute one mechanism of adaptation to the farm environment.

Flexible chemotherapy regimen with gemcitabine and vinorelbine for metastatic nonsmall cell lung carcinoma: a phase II multicenter trial.

BACKGROUND: This Phase II study evaluated a flexible 3- or 4-week dosing schedule of gemcitabine and vinorelbine to determine its effect on response rate and survival of patients with metastatic nonsmall cell lung carcinoma (NSCLC). METHODS: Thirty-four response-evaluable patients, 24 with performance status (PS) 0-1 and 10 with a PS of 2, 30 with Stage IV, and 4 with Stage IIIB NSCLC were treated with gemcitabine 1000 mg/m(2) intravenously and vinorelbine 25 mg/m(2) intravenously (first 15 patients) or 30 mg/m(2) intravenously (next 19 patients) on Days 1, 8, and 15 of a 4-week cycle, if on Day 15 neutrophils were > or = 1500/uL and platelets > or = 100,000/uL. If chemotherapy could not be administered on Day 15, then Day 22 became Day 1 of the next cycle. RESULTS: When vinorelbine 25 mg/m(2) was given with gemcitabine 1000 mg/m(2), 11 patients received 4-week cycles, 3 patients 3-week cycles, and 1 patient both 3- and 4-week cycles. With vinorelbine 30 mg/m(2) and gemcitabine 1000 mg/m(2), 7 patients received 4-week cycles, 2 patients 3-week cycles, and 10 patients both 3- and 4-week cycles. The partial response rate for 34 patients was 53% (18 patients). Median survival (MS) was 11.1 months, and 1-year survival 50% (17 patients). Patients with PS 0+1 had a MS of 17.5 months compared with patients with PS 2, who had MS of 3.3 months. Patients < 70 years of age had a MS of 18 months, and those >/= 70 years had a MS of 5.5 months. CONCLUSION: This flexible schedule with gemcitabine and vinorelbine enabled optimal dose delivery and suggested excellent efficacy but less toxicity than treatment with platinum regimens.

Random amplified ribosomal DNA restriction analysis for rapid identification of thermophilic Actinomycete-like bacteria involved in hypersensitivity pneumonitis.

Hypersensitivity pneumonitis (HP) is a pulmonary disease characterised by inflammation that can be caused by, amongst other substances, a subset of 4 thermophilic mycelial bacteria: Saccharopolyspora rectivirgula, Saccharomonospora viridis, Thermoactinomyces sacchari, and Thermoactinomyces vulgaris. Air sampling analyses in highly contaminated environments are often performed to evaluate exposure to these species which are difficult and fastidious to identify by conventional techniques. The aim of this study was to use amplified ribosomal DNA restriction analysis (ARDRA) to develop a method of identification for those thermophilic organisms that would be more rapid and simple. Strains of these 4 species were obtained from the American type culture collection (ATCC) and were characterized using biochemical tests and ARDRA patterns obtained on their partial-lenght amplified 16S rDNAs. To validate this approach, ARDRA with two restriction enzymes, TaqI and HhaI, was applied to 49 thermophilic actinomycete-like strains from environmental samples (sawmills). The results obtained show that combining some cultural characteristics and biochemical tests, such as xanthine or hypoxanthine decomposition, growth in the presence of NaCl, lysozyme or novobiocin, and spore resistance over 100 degrees C provide a rough identification and selection of the genera of interest. Consequently, target species could be confirmed by digestion of partial-lenght 16S rDNA with the use of Taql and HhaI restriction enzymes that gave specific restriction patterns. ARDRA analyses on the 49 environmental actinomycete-like organisms revealed the presence of 8 Saccharopolyspora rectivirgula, 2 Saccharomonospora viridis, and 15 Thermoactinomyces vulgaris strains, the other strains had restriction patterns different than those of the species of interest. Results of the present study will be applicable to other potential HP environments such as dairy barns, peat bogs and compost plants.

Comparison of endotoxin exposure assessment by bioaerosol impinger and filter-sampling methods.

Environmental assessment data collected in two prior occupational hygiene studies of swine barns and sawmills allowed the comparison of concurrent, triplicate, side-by-side endotoxin measurements using air sampling filters and bioaerosol impingers. Endotoxin concentrations in impinger solutions and filter eluates were assayed using the Limulus amebocyte lysate assay. In sawmills, impinger sampling yielded significantly higher endotoxin concentration measurements and lower variances than filter sampling with IOM inhalable dust samplers. Analysis of variance for repeated measures showed that this association remained after controlling for other factors such as replicate, sawmill, sawmill operation, wood type, and interaction terms. Endotoxin concentrations in the swine barns were 10-fold higher on average than in sawmills. These samples demonstrated comparable endotoxin concentration estimates for impinger and filter methods although the variability was lower using the impinger method. In both occupational settings, side-by-side replicates were more uniform for the impinger samples than for the filter samples. This study demonstrates that impinger sampling is an acceptable method for quantitation of area endotoxin concentrations. Further, when sampling is performed with impingers for airborne microorganism quantitation, these same impinger solutions can yield valid endotoxin exposure estimates, negating the need for additional filter sampling.

Respiratory health impact of working in sawmills in eastern Canada.

Air contamination in sawmills can cause respiratory health problems. The authors measured respirable dust, bacteria, endotoxins, and molds collected from 17 sawmills in eastern Canada. A total of 1,205 sawmill workers answered a respiratory-health questionnaire, and they all participated in lung-function measurements, skin-prick tests, and venous blood sampling for specific immunoglobulins against molds found in the sawmills. Workers had normal lung functions, and most respiratory symptoms could be explained by smoking histories. Workers in pine sawmills had a greater prevalence of positive skin-prick test to pine than did workers in sawmills where other woods were used. High levels of specific antibodies were seen in some workers. The presence of a positive skin-prick test and/or specific antibodies had no impact on lung function(s). These Quebec sawmill workers did not experience significant respiratory illnesses; however, some of these workers may be at a higher risk of developing asthma and hypersensitivity pneumonitis than nonworkers.

Assessment of particulates and bioaerosols in eastern Canadian sawmills.

The purpose of this study was to quantify and identify the airborne contamination in eastern Canadian sawmills. Seventeen sawmills were chosen to cover a wide range of size, geographic distribution, and wood species processed. Within each sawmill different work sites (debarking, sawing, sorting, or planing) were studied separately. Area sampling was performed for exposure assessment. Microbial contaminants were assessed with all-glass impingers 30 and six-stage Andersen microbial samplers; appropriate selective media and culture conditions for bacteria, thermophilic actinomycetes, molds, and yeasts were used. Inhalable dust, endotoxins, temperature, and humidity also were measured. Penicillium species were the most predominant molds with up to 40 different Penicillium species identified. Debarking was the working site most highly contaminated by molds, bacteria, and endotoxins (p=0.0001). At this working site mold levels reached a maximum of 1.5 x 10(6) CFU/m3, whereas the median values for culturable bacteria and endotoxin were 21,620 CFU/m3 and 1,081 endotoxin units/m3, respectively. Planing sites were the most highly dust contaminated (median: 3.0 mg/m3) (p <0.05). Sawmills of eastern Canada contain airborne biological contaminants that vary between working sites, and their microflora is different from that previously described in European sawmills.

Granzyme activity in the inflamed lung is not controlled by endogenous serine proteinase inhibitors.

Numerous lung diseases, such as hypersensitivity pneumonitis (HP), are characterized by the presence of activated alveolar CTL and NK cells. Since these cells produce granzymes, granzyme A and B levels in bronchoalveolar lavage (BAL) fluids from 14 normal subjects and 12 patients with HP were measured by ELISA. Median (range) BAL granzyme A and B levels were 4 (0-37) and 0 (0-6) pg/ml in normal subjects. BAL granzyme levels were significantly higher in HP patients, being at 74 (0-1,889) and 10 (0-78) pg/ml for granzymes A and B, respectively. In vitro, neither of the three main serine protease inhibitors of the lung, namely alpha1-antitrypsin, secretory leukocyte protease inhibitor, and elafin, showed any effect on granzyme A or B activity. In addition, granzyme A was shown to be fully active in BAL fluids. Hence, these data show that granzyme activity may be poorly controlled by protease inhibitors in inflamed tissues. Thus, granzymes could contribute to tissue remodeling and inflammation characterizing HP.