5-lipoxygenase and cyclooxygenase-2 in porcine parasitic bronchopneumonia: immunohistochemical and biochemical investigations.

Eicosanoids are products of arachidonic acid metabolism and have numerous biological roles. The present study aimed to investigate the role of 5-lipoxygenase (5-LOX)- and cyclooxygenase-2 (COX-2)- dependent enzymatic pathways in the pathogenesis of porcine parasitic bronchopneumonia caused by Metastrongylus spp. Pulmonary tissue samples from healthy control and parasitized pigs were processed for histopathological, immunohistochemical and biochemical investigations. In control animals, immunohistochemistry demonstrated that 5-LOX and COX-2 expression was almost exclusively limited to the bronchiolar epithelial cells. Parasitized pigs had greater 5-LOX- and COX-2- specific immunoreactivity, involving a wide range of cell types within foci of granulomatous and eosinophilic bronchopneumonia. Biochemical investigations demonstrated the presence of 5-LOX (and the related product Leukotriene B(4)) and COX-2 (and the related product prostaglandin E(2); PGE(2)) in all tissues under study. COX-2 activity and PGE(2) concentration were significantly higher in diseased lungs compared with normal healthy controls. These findings demonstrate that 5-LOX and COX-2 are differentially expressed in normal versus lungworm-infected lungs and therefore suggest that both biochemical pathways are likely to be involved in the pathogenesis of porcine parasitic bronchopneumonia.

Expression of secretory phospholipase A2 enzymes in lungs of humans with pneumonia and their potential prostaglandin-synthetic function in human lung-derived cells.

Although a number of sPLA2 (secretory phospholipase A2) enzymes have been identified in mammals, the localization and functions of individual enzymes in human pathologic tissues still remain obscure. In the present study, we have examined the expression and function of sPLA2s in human lung-derived cells and in human lungs with pneumonia. Group IID, V and X sPLA2s were expressed in cultured human bronchial epithelial cells (BEAS-2B) and normal human pulmonary fibroblasts with distinct requirement for cytokines (interleukin-1b, tumour necrosis factor a and interferon-g). Lentivirus- or adenovirus-mediated transfection of various sPLA2s into BEAS-2B or normal human pulmonary fibroblast cells revealed that group V and X sPLA2s increased arachidonate release and prostaglandin production in both cell types, whereas group IIA and IID sPLA2s failed to do so. Immunohistochemistry of human lungs with pneumonia demonstrated that group V and X sPLA2s were widely expressed in the airway epithelium, interstitium and alveolar macrophages, in which group IID sPLA2 was also positive, whereas group IIA sPLA2 was restricted to the pulmonary arterial smooth muscle layers and bronchial chondrocytes, and group IIE and IIF sPLA2s were minimally detected. These results suggest that group V and X sPLA2s affect lung pathogenesis by facilitating arachidonate metabolism or possibly through other functions.

In vitro and in vivo activities of meropenem and comparable antimicrobial agents against Haemophilus influenzae, including beta-lactamase-negative ampicillin-resistant strains.

The in vitro activity of ampicillin, cefotaxime, meropenem, panipenem, imipenem and biapenem was assayed using ampicillin-susceptible, beta-lactamase-positive and beta-lactamase-negative ampicillin-resistant (BLNAR) Haemophilus influenzae isolated recently in Japan. Against ampicillin-susceptible isolates, cefotaxime was the most potent (MIC(90) 0.016 mg/mL). Both cefotaxime and meropenem (MIC(90) of both, 0.5 mg/L) were the most potent against beta-lactamase-positive isolates. Against BLNAR isolates, meropenem (MIC(90) 0.5 mg/L) was the most potent. In murine bronchopneumonia caused by ampicillin-susceptible and BLNAR H. influenzae, cefotaxime showed the best efficacy, followed by meropenem. Our results indicate that meropenem could be a useful intravenous agent for infections caused by H. influenzae, including BLNAR strains.

Specific enzyme activities in bronchoalveolar lavage fluid as an aid to diagnosis of tracheobronchitis and bronchopneumonia in dogs.

Lactate dehydrogenase (LDH), alkaline phosphatase (ALP), alanine aminotransferase and gamma-glutamyl transferase enzyme activities, and total protein (TP), calcium, inorganic phosphate, urea nitrogen (UN) and creatinine concentrations in bronchoalveolar lavage fluid were investigated for their relative importance in the diagnosis of respiratory diseases in dogs. Bronchoalveolar lavage (BAL) fluid was obtained from 26 dogs (20 with respiratory diseases and six controls) following anaesthesia with sodium pentothal. Enzyme activities and biochemical parameters were measured in BAL fluid. LDH and ALP levels were significantly increased in 12 dogs with bronchopneumonia, but not in eight dogs with tracheobronchitis. Insignificant and variable levels of TP and UN concentrations were found in both groups. It was concluded that LDH and ALP enzyme activities could be considered as pointers to pulmonary inflammation and/or damage while TP and UN measurements in BAL fluid may have a place in the identification of changes in respiratory and vascular permeability.

Expression of inducible nitric oxide synthase in spontaneous bovine bronchopneumonia.

The expression of inducible nitric oxide synthase (iNOS), major histocompatibility class II molecules (MHC-II), CD68, and the calcium-binding proteins S100A8 and S100A9 (also called MRP8 and MRP14, respectively) was assessed in lung tissues from cattle that succumbed to pneumonia. Expression patterns of these markers were related to the types of lung lesion. iNOS expression was only observed in lungs infected with Arcanobacterium pyogenes or Pasteurella haemolytica but not in lungs from cattle with subacute chronic interstitial pneumonia and acute interstitial pneumonia due to Escherichia coli infection. High levels of iNOS were expressed by cells (probably leukocytes) surrounding necrotic foci. Occasionally, iNOS was expressed by intraalveolar macrophages in viable parenchyma, by leukocytes within the airways, and by some chondrocytes in the supporting cartilage of bronchi. Cells expressing MHC-II were distributed relatively evenly throughout areas of inflammation and did not display any clear association with necrotic foci. Cell types expressing MHC-II included type II alveolar epithelial cells, spindle-shaped cells of the interstitium, cells in bronchus-associated lymphoid tissue, and leukocytes in lymph and blood vessels but largely excluded iNOS-positive cells. Likewise, CD68-positive cells were rarely positive for iNOS and were not confined to the areas surrounding necrotic tissue. As with MHC-II and CD68, there was little if any coexpression of iNOS and either of the S100 proteins tested. Thus, in cattle with necrotizing bronchopneumonia, iNOS-expressing cells were largely restricted to the cellular zone surrounding necrotic areas.

A morphologic study of lung secretory leukoprotease inhibitor in pneumonia.

The present study was undertaken to determine the localization of cells laden with secretory leukoprotease inhibitor (SLPI) in bronchial/bronchiolar epithelium (B/Br-E) by histochemical techniques to see whether SLPI production occurs in conjunction with pathologic bronchopneumonia. Ten lung were obtained at autopsy from patients between 63 and 100 yr of age, including six with pathologic pneumonia and four without pneumonia. SLPI-laden cells in the B/Br-E corresponded mostly to goblet cells with apparent hyperplasia. A morphometric study performed on the B/Br-E indicated that the percentage of SLPI-laden cells was significantly correlated with the percentage of mucus-containing cells (r = 0.72, p < 0.001). This trend was similar in the bronchi (r = 0.60, p < 0.05) and in the bronchioles (r = 0.90, p < 0.01). The increased percentage of mucus and SLPI-laden cells in the B/Br-E was closely correlated with acute inflammatory changes in the adjacent alveoli, particularly in bronchi rather than in bronchioles. From these observations we conclude that the number of SLPI-laden cells in the airways increases in correlation with goblet cell hyperplasia. In addition, these morphologic changes are associated with the existence of acute inflammatory cell infiltration in the alveolar area.

Inositol-specific phospholipase D activity in health and disease.

1. We report the first demonstration of the pathophysiological importance and clinical applications of the relatively recently discovered circulating enzyme, phosphoinositol-specific phospholipase D. This enzyme is known to cleave the large variety of important cell-surface molecules linked to the cell membrane by glycan-phosphatidylinositol linkages (glycan-phosphatidylinositol anchors). 2. When measured in the sera of healthy individuals, phosphoinositol-specific phospholipase D activity was found to show a strong negative correlation with age, the degree of depreciation being greater than that measured for most other analytes. 3. Serum phosphoinositol-specific phospholipase D activity was considerably depressed in patients presenting with conditions leading to reduced liver synthetic reserve, such as hepatocellular carcinoma or liver cirrhosis caused by chronic viral hepatitis, and correlated with reduced albumin levels in these conditions, indicating that the liver is the site of phosphoinositol-specific phospholipase D synthesis and that phosphoinositol-specific phospholipase D may be used as an additional marker of liver synthetic reserve. 4. When measured in patients with acute liver disease, such as acute viral hepatitis, or in patients with bronchopneumonia, phosphoinositol-specific phospholipase D activity was found to be significantly raised, demonstrating features characteristic of an acute-phase reactant. 5. These findings indicate that, besides its pathophysiological importance, phosphoinositol-specific phospholipase D and the measurement of its activity in serum may have a useful place in the investigation of a range of clinical conditions, including tissue injury and inflammation.

Human neutrophil elastase: degradation of basement membrane components and immunolocalization in the tissue.

Human neutrophil elastase was purified to homogeneity as two isozymes named E1 and E2. The isozymes degraded Type IV collagen, laminin, fibronectin, and heparan sulfate proteoglycan similarly to each other. The degradation of such basement membrane components by elastase may assist the extravasation of neutrophils in the process of inflammation. Among the substrates tested, only type V collagen, which is susceptible to neutrophil gelatinase, was resistant to elastase. This broad substrate specificity of the enzyme may also contribute to tissue destruction at the sites of inflammation. We produced a monoclonal antibody against the purified enzyme and applied it to immunohistochemical studies. In bronchopneumonia and polyarteritis nodosa, elastase was associated with the cleaved elastic fibers, indicating that the enzyme really destroys tissue in vivo. In the exudates of rheumatoid joint, elastase was stained as diffuse fine granules. Immunohistochemical studies with the monoclonal antibody will provide a complementary way to disclose the mechanism of diseases related to neutrophil infiltration.

[Lysozyme content in biological substrates and immunological indices of the blood in pneumonia patients]. / Soderzhanie lizotsima v biologicheskikh substratakh i immunologicheskie pokazateli krovi bol'nykh pnevmoniei.

An increase of lysozyme content in biological fluids observed during treatment of pulmonary diseases was shown to be followed by positive immunological and clinical effects. The parameters of immunological properties of lysozyme determined on the patient blood cells in vitro (before treatment) may be used for prediction of the drug efficacy during treatment.

[Phospholipid biosynthesis in the lungs in a chronic inflammatory bronchopulmonary process]. / Biosintez fosfolipidov v legkikh pri khronicheskom vospalitel'nom bronkholegochnom protsesse.

Glycerokinase, glycerophosphate dehydrogenase, glycerophosphate acyltransferase activity, glycerophosphate, dioxiacetonphosphate level and in vivo incorporation of (U-14C)-glucose into the lung phospholipid structure were studied in normal rats and in conditions of chronic bronchopulmonary inflammation. The inhibition of glycerokinase and glycolytic ways of glycerophosphate formation was demonstrated. The data obtained have shown that glucose incorporation into phosphatidyl cholines, phosphatidyl glycerols and phosphatidyl ethanolamines was decreased, while the incorporation of radioactivity into the sphingomyelins and lysophosphatidyl cholines was significantly activated.

[Local and general changes in inflammatory lung diseases]. / Mestnye i obshchie izmeneniia pri vospalitel'nykh zabolevaniiakh legkikh.

Composite histo-cytoenzymochemical characteristics of local (pulmonary tissue) and general (polymorphonuclear leukocytes, PMNL, blood lymphocytes) changes in inflammatory lung diseases were obtained. Local and general changes were found to be interrelated and to reflect morphological and clinical features of inflammatory lung diseases. Differential diagnosis between acute and protracted bronchopneumonia, chronic pneumonia, bronchoectases is possible on the basis of the condition of PMNL and blood lymphocytes.

A contribution to study of acute obstructive bronchitis and bronchopneumonia in infants.

103 infants with acute obstructive bronchitis and/or bronchopneumonia were studied. IgM and alpha-1-antitrypsin values were determined by single radial immunodiffusion. In each infant the values were studies in eight serum samples. Comparison of the time of clinical recovery and x-ray clearing of the lung and the time of normalization of IgM and alpha-1-antitrypsin was also done. In infants with acute obstructive bronchitis the clinical recovery and X-ray clearing occurred on about the 13th day from the beginning of investigation. However, the normalization of IgM and alpha-1-antitrypsin values, which were significantly increased in all investigated serum samples, did not occur even on the 44th day of investigation. In infants with acute obstructive bronchitis associated with bronchopneumonia the clinical recovery occurred on the 10th day of investigation and X-ray clearing on the 14th day. IgM and alpha-1-antitrypsin values, which were also significantly increased in all investigated serum samples, were normalized on the 44th day of investigation. In group of infants with bronchopneumonia the clinical recovery occurred, approximatively, on the 12th day, whole the x-ray clearing occurred on the 17th day of investigation. IgM and alpha-1-antitrypsin values were normalized on the 18th day of investigation. In infants with acute obstructive bronchitis and/or bronchopneumonia there is significant correlation between IgM and alpha-1-antitrypsin values. This finding could be used to evaluate which of these two disease is dominant if they occur at the same time in infant.

Glycolytic enzymes from human autoptic brain cortex: normal aged and demented cases.

The activities of glycolytic enzymes were determined in human autoptic temporal lobes from patients with different forms of dementia. For some enzymes (hexokinase, phosphofructokinase and phosphoglycerate mutase) the effect seen in dementia can be regarded as an intensification of the normal ageing affect. For other enzymes (aldolase, phosphoglucose isomerase, triosephosphate isomerase and lactate dehydrogenase) no changes in enzyme activities corresponding to those found in dementia are observed in the normal ageing process. These effects are most pronounced in the non-vascular Alzheimer cases. With the exception of triosephosphate isomerase and lactate dehydrogenase, enzyme activity is also reduced in bronchopneumonia. The effects of dementia and bronchopneumonia on the activities of glycolytic enzymes in human autoptic brain tissue are often difficult to distinguish.

Choline acetyltransferase and acetylcholinesterase abnormalities in senile dementia: importance of biochemical measurements in human post-mortem brain specimens.

Choline acetyltransferase and acetylcholinesterase have been assessed in human aging brains, in demented and agonal states. Choline acetyl transferase decreased during aging in normal brain when measured in the cerebral cortex. Choline acetyltransferase was also reduced in several other brain areas in patients with Alzheimer's disease and in one patient with Creutzfeldt-Jakob disease. Choline acetyltransferase was also reduced in bronchopneumonia and in some terminal conditions. On the contrary, the activity was not reduced in patients who died after cerebrovascular accidents. Acetylcholine esterase, although it followed the general trend of choline acetyltransferase, did not yield significant results.

Neurotransmitter-related enzymes and indices of hypoxia in senile dementia and other abiotrophies.

Fifty-six brains from middle-aged and elderly normal as well as demented subjects and patients with provisional clinical diagnosis of other neurological and psychiatric diseases were assessed histologically. On this basis the specimens were classified into 14 diagnostic groups. A survey of potential indices of specific neurons has been carried out on these brains in which neurotransmitter-related enzymes, gamma-GTP (a potential index of capillaries) and specific proteins have been determined in up to 20 brain regions. In addition, the agonal state has been tentatively assessed by examining the post-mortem states of the circulatory and respiratory systems. CAT and gamma-GTP activities and the concentration of a soluble neuronal-type protein (neuronin S-5) were found to be relatively unaffected by the agonal state. When cases of senile dementia were compared to controls (matched with respect to the cause of death) the activity of CAT (the potential index of cholinergic neurons) appears to be reduced in the cerebral cortex. This is a preliminary finding, although a correlation was indicated between CAT activity and 'senile' morphological changes, the activity was markedly reduced in only 3 brains. However, despite inconsistencies in the literature (Karczmar, 1975) at least one pharmacological study on humans appears to show that the cholinergic system may be involved in age-related memory degeneration (Drachman and Leavitt, 1974). Cholinergic neurons may be abnormal in the other abiotrophies examined (Huntington's chorea, motor neuron disease and mixed vascular and senile dementia). gamma-GTP and neuronin S-5 (identical in most respects to the soluble acidic neuronal protein 14-3-2 of antigen alpha) were not reduced in senile dementia. The activities of brain decarboxylase (GAD and AAD) and the concentration of another soluble acidic brain protein (neuronin S-6) appear to be affected by the agonal state. This is remarkable because GAD and, in particular, neuronin S6, are relatively unaffected by post-mortem autolysis. As judged by the state of the extraneural systems which regulated the blood and oxygen supply to the brain it appears that terminal 'cerebral hypoxia' is responsible for the depletion of these brain constituents. This effect appears to be particularly marked in deep grey matter. In non-demented patients that die of bronchopneumonia, the areas of the cortex which are depleted in neuronin S-6 are consistent with the pattern of the 'selective vulnerability' of the cortex to hypoxia, suggesting that the terminal state can also affect the neocortex. If so, then this is particularly relevant to studies on senile dementia, for the effect of the terminal bronchopneumonia that so often occurs in these patients (and in patients with other abiotrophies) may be exacerbated by a terminal reduction in cerebral blood flow...

[Various pulmonary lesions induced by high-pressure oxygen]. / Osservazioni relative ad alcune lesioni polmonari indotte dall'ossigeno a forti pressioni

Hyperbaric oxygen may provoke lesions in various organs and tissues depending on the dose and application time. The toxic action mechanisms of oxygen are manifold. The pulmonary lesions that occur in mice submitted to various oxygen pressures are investigated. Anatomo-pathological examination revealed numerous alterations of various kinds depending on pressure used. Emphysema, pulmonary oedema and enormous inflammatory processes in the lung are the most frequent findings encountered in research.