Trichinella spiralis infection enhances protein kinase C phosphorylation in guinea pig alveolar macrophages.

To learn more about the signalling pathways involved in superoxide anion production in guinea pig alveolar macrophages, triggered by Trichinella spiralis infection, protein level and phosphorylation of mitogen activated protein (MAP) kinases and protein kinase C (PKC) were investigated. Infection with T. spiralis, the nematode having 'lung phase' during colonization of the host, enhances PKC phosphorylation in guinea pig alveolar macrophages. Isoenzymes beta and delta of PKC have been found significantly phosphorylated, although their location was not changed as a consequence of T. spiralis infection. Neither in macrophages from T. spiralis-infected guinea pig nor in platelet-activating factor (PAF)-stimulated macrophages from uninfected animals, participation of MAP kinases in respiratory burst activation was statistically significant. The parasite antigens seem to act through macrophage PAF receptors, transducing a signal for enhanced NADPH oxidase activity, as stimulating effect of newborn larvae homogenate on respiratory burst was abolished by specific PAF receptor antagonist CV 6209. A suppressive action of T. spiralis larvae on host alveolar macrophage innate immunological response was reflected by diminished protein level of ERK2 kinase and suppressed superoxide anion production, in spite of high level of PKC phosphorylation.

Developmental arrest in Caenorhabditis elegans dauer larvae causes high expression of enzymes involved in thymidylate biosynthesis, similar to that found in Trichinella muscle larvae.

Crude extract specific activities of thymidylate synthase, dUTPase, thymidine kinase and dihydrofolate reductase were high during the development of Caenorhabditis elegans, the dauer larva activities being similar to those previously determined in Trichinella spiralis and T. pseudospiralis muscle larvae (with the exception of thymidine kinase, not detected in Trichinella). High thymidylate synthase expression in developmentally arrested larvae, demonstrated also at the mRNA and protein levels, is in agreement with a global cell cycle arrest of dauer larvae and indicates this unusual cell cycle regulation pattern can be shared by developmentally arrested larvae of C. elegans and the two Trichnella species. Hence, the phenomenon may be characteristic for developmentally arrested larvae of different nematodes, rather than specific for the parasitic Trichinella muscle larvae. Endogenous C. elegans thymidylate synthase was purified and its molecular properties compared with those of the recombinant protein, expression of the latter in E. coli cells confirming the NCBI database sequence identity.

A non-classical type of alveolar macrophage response to Trichinella spiralis infection.

Studies of arginase expression and activity in guinea pig alveolar macrophages during Trichinella spiralis infection, prompted by earlier observation of innate lung response to the parasite, showed the macrophages to express both activity and protein of arginase type I. In cultured macrophages part of the enzyme was found to be always released to the extracellular medium. Whereas BCG in vivo treatment, alone or preceded by T. spiralis infection, stimulated arginase activity, T. spiralis infection alone affected the enzyme distribution between intracellular and extracellular fractions, and properties (K(m) and V(max)), rather than total (intracellular + extracellular) activity, with TGF-beta apparently responsible for a part of the effect. Anti-TGF-beta antibody treatment of the animals influenced both arginase activation by Mn(2+) and dependence of the enzyme-catalysed reaction on pH. Whereas T. spiralis infection activated guinea pig alveolar macrophages by the type II macrophage activation, as indicated by constant arginase expression, associated with previously demonstrated lack of stimulation of nitric oxide production, BCG treatment invoked an alternative type of activation mechanism, reflected by stimulation of macrophage arginase, but not iNOS, activity.

[Extracellular components of bronchoalveolar lavage fluid (BALF) as a marker of the activity of interstitial pulmonary diseases. III: Phospholipids to protein concentration ratio]. / Pozakomórkowe skladniki plynu z plukania oskrzelowo-pçherzykowego (BALF) jako marker aktywnosci sródmiazszowych chorób pluc. Czesc III: Stosunek stezenia fosfolipidów do stezenia bialka.

In interstitial pulmonary diseases investigations are conducted to find markers of the activity of the interstitial processes so that noninvasive monitoring of the disease might be possible. In 188 patients divided into 9 groups: 42 with active sarcoidosis, 24 with inactive sarcoidosis, 16 with active sarcoidosis treated with steroids and 22 with inactive sarcoidosis after corticotherapy, 17 with avian fanciers' lung exposed to the antigen, 16 with avian fanciers' lung after a year interval in exposure to the antigen, 20 with advanced and 13 with moderate idiopathic pulmonary fibrosis, and 18 healthy persons the BAL was performed. In the BALF concentrations of protein and phospholipids were assayed by colorimetric method. The results indicate usefulness of the studied biochemical parameters in BALF in evaluation of the activity of interstitial pulmonary diseases. Significant differences were found between the results in the active group of patients compared to the control group and to the inactive forms of interstitial pulmonary diseases. Particularly valuable is phospholipids to protein concentration ratio in BALF.

Obesity induces expression of uncoupling protein-2 in hepatocytes and promotes liver ATP depletion.

Uncoupling protein 2 (UCP2) uncouples respiration from oxidative phosphorylation and may contribute to obesity through effects on energy metabolism. Because basal metabolic rate is decreased in obesity, UCP2 expression is predicted to be reduced. Paradoxically, hepatic expression of UCP2 mRNA is increased in genetically obese (ob/ob) mice. In situ hybridization and immunohistochemical analysis of ob/ob livers demonstrate that UCP2 mRNA and protein expression are increased in hepatocytes, which do not express UCP2 in lean mice. Mitochondria isolated from ob/ob livers exhibit an increased rate of H+ leak which partially dissipates the mitochondrial membrane potential when the rate of electron transport is suppressed. In addition, hepatic ATP stores are reduced and these livers are more vulnerable to necrosis after transient hepatic ischemia. Hence, hepatocytes adapt to obesity by up-regulating UCP2. However, because this decreases the efficiency of energy trapping, the cells become vulnerable to ATP depletion when energy needs increase acutely.

[Extracellular components of broncho-alveolar lavage fluid (BALF) as markers of interstitial pulmonary disease activity. II. Phospholipid concentration]. / Pozakomórkowe skladniki plynu z plukania oskrzelowo-pecherzykowego (BALF) jako marker aktywnosci sródmiazszowych chorób pluc. Czesc II: Stezenie fosfolipidów.

Phospholipids found in the alveolar space come from the surfactant which is produced by type II pneumocytes and whose surplus is eliminated from lungs by macrophages. In 188 patients with interstitial pulmonary diseases bronchoalveolar lavage was performed. Phospholipids from the supernatant of BALF were extracted using of Folch method and their concentration was measured colorimetrically assaying inorganic phosphorus after prior mineralization by Fiske-Subbarov method. The persons were divided into 7 groups: active sarcoidosis, inactive sarcoidosis, avian fancier's lung in the period of contact with the antigen, and after stopping the contact with the antigen, advanced idiopathic pulmonary fibrosis, moderate idiopathic pulmonary fibrosis, and a control group of healthy persons. Apart from that, the patients were divided into untreated and treated with corticosteroids. In every group of patients we have noticed increased concentration and total amount of phospholipids in BALF. Particularly distinct increase of the total number of phospholipids in BALF in pulmonary fibrosis was observed. However, this parameter does not permit to estimate disease activity in interstitial lung disease. Corticotherapy increases phospholipids concentration in BALF in patients with idiopathic pulmonary fibrosis and avian fancier's lung but decreases in sarcoidosis patients.

[Extracellular components of bronchoalveolar lavage fluid (BALF) as a marker of interstitial pulmonary disease activity. I. Protein concentration]. / Pozakomórkowe skladniki plynu z plukania oskrzelowo-pecherzykowego (BALF) jako marker aktywnosci sródmiazszowych chorób pluc. Czesc I. Stezenie bialka.

In interstitial pulmonary diseases a passive transudation of protein into an alveolar space takes place as well as local synthesis of protein by cells in alveolar space and protein leaking out of destroyed cells. Concentration of protein in BALF was assessed in 170 patients with interstitial pulmonary diseases and in 18 healthy persons by means of colorimetric method with Coomassie Blue. Patients were divided into 8 groups: active untreated sarcoidosis, inactive untreated sarcoidosis, active sarcoidosis after corticotherapy, inactive sarcoidosis after corticotherapy, avian fanciers' lung in the period of contact with the antigen, avian fanciers' lung after one year's interval in exposure to the antigen, advanced idiopathic pulmonary fibrosis, and moderate idiopathic pulmonary fibrosis. In the group with active forms of diseases the increase of protein concentration in BALF was observed. The increase was statistically significant compared with the results in the control group and in the groups with inactive forms of the diseases. Protein concentration in BALF may play the role as marker of activity in interstitial pulmonary diseases.

Hormonal stimulation, mitochondrial Ca2+ accumulation, and the control of the mitochondrial permeability transition in intact hepatocytes.

Ca2+ functions as an intracellular signal to transfer hormonal messages to different cellular compartments, including mitochondria, where it activates intramitochondrial Ca2(+)-dependent enzymes. However, excessive mitochondrial Ca2+ uptake can promote the mitochondrial permeability transition (MPT), a process known to be associated with cell injury. The factors controlling mitochondrial Ca2+ uptake and release in intact cells are poorly understood. In this paper, we investigate mitochondrial Ca2+ accumulation in intact hepatocytes in response to the elevation of cytosolic Ca2+ levels ([Ca2+]c) induced either by a hormonal stimulus (vasopressin), or by thapsigargin, an inhibitor of the endoplasmic reticulum Ca2+ pump. After stimulation, cells were rapidly permeabilized for the determination of the mitochondrial Ca2+ content (Ca2+(m)) and to analyze the susceptibility of the mitochondria to undergo the MPT. Despite very similar levels of [Ca2+]c elevation, vasopressin and thapsigargin had markedly different effects on mitochondrial Ca2+ accumulation. Vasopressin caused a rapid (< 90 sec), but modest (< 2 fold) increase in Ca2+(m) that was not further increased during prolonged incubations, despite a sustained [Ca2+]c elevation. By contrast, thapsigargin induced a net Ca2+ accumulation in mitochondria that continued for up to 30 min and reached Ca2+(m) levels 10-20 fold over basal. Accumulation of mitochondrial Ca2+ was accompanied by a markedly increased susceptibility to undergo the MPT. Both mitochondrial Ca2+ accumulation and MPT activation were modulated by treatment of the cells with inhibitors of protein kinases and phosphatases. The results indicate that net mitochondrial Ca2+ uptake in response to hormonal stimulation is regulated by processes that depend on protein kinase activation. These controls are inoperative when the cytosol is flooded by Ca2+ through artificial means, enabling mitochondria to function as a Ca2+ sink under these conditions.

[Concentration of thromboxane B2 in bronchoalveolar lavage fluid in selected interstitial pulmonary diseases]. / Stezenie tromboksanu B2 w plynie z plukania oskrzelowo-pecherzykowego w wybranych chorobach sródmiazszowych pluc.

Thromboxane B2 (TxB2) is believed to play some role in pathogenesis of interstitial pulmonary diseases. The concentration of TxB2 in BALF was assayed in 75 ill and 6 healthy persons. The ill were divided into 6 groups: with active sarcoidosis [18], with active sarcoidosis treated with corticosteroids [8], with advanced pulmonary fibrosis [15], with moderate pulmonary fibrosis [6], with avian fanciers lung after exposition of antigen [16], avian fanciers lung without exposition of antigen [13]. Radioimmunological method with the Amersham set to assay the concentration of TxB, was used. A rise of TxB2 concentration was detected in the group with advanced pulmonary fibrosis. The rise was significant compared with the group of healthy persons, those with sarcoidosis, and with moderate pulmonary fibrosis. A tendency to higher concentration of TxB2 in patients with avian fanciers lung exposed to the allergen was also observed. The results indicate that TxB2 takes part in pathogenesis of these interstitial pulmonary diseases in which increased number of neutrophils in BALF is found.

[Concentration of sialic acid in bronchoalveolar lavage fluid in selected interstitial pulmonary diseases]. / Stezenie kwasu sialowego w plynie z plukania oskrzelowo-pecherzykowego w wybranych chorobach sródmiazszowych pluc.

Sialic acid is a component of cell membranes and it can take part in immunological processes with lymphocytes and neutrophiles. In the present work the concentration of sialic acid in BALF in selected interstitial pulmonary disease was studied (in sarcoidosis, pulmonary fibrosis, avian fanciers lung) and compared with the control group of healthy persons. The investigations were repeated during the observations. The patients were divided into active and inactive groups. The analysis included the results in 187 patients divided into 9 groups. Sialic acid was measured with colorimetric method with the use of Ehrlich's reagent. The growth of sialic acid concentration in supernatant of BALF was observed in the studied diseases. The increase of sialic acid occurs in inflammatory processes especially those with neutrophiles. The correlation of acid concentration with the percentage or the total number of lymphocytes in BALF was observed. It is specific that the concentration of sialic acid in BALF in patients with avian fanciers lung keeps growing even after the contact with the antigen was discontinued.

Effect of hyperoxia on histamine metabolism in rat brain synaptosomes: preliminary observations.

Adult male Wistar rats were submitted to normobaric hyperoxygenation for 1 and 4 hours, then brain synaptosomes were isolated and uptake and release of the histamine precursor - histidine (His), histamine (HA) level and His metabolizing enzymes activities were measured. This uptake in hyperoxic synaptosomes was inhibited by about 20%. After 1-hour hyperoxia, a tendency towards an increase of the HA level, but a significant increase histidine decarboxylase (HD) and histamine methyltransferase (HMT) activities were observed. Four-hour hyperoxia caused a decrease of both the HA level and the activities of both enzymes, especially HMT. The changes were reversed in 1-hour posthyperoxic recovery, except for histidine uptake which remained inhibited.

In vitro and in vivo effect of salbutamol on histamine release from human basophils.

The effect of salbutamol, a selective beta 2-adrenergic agonist, on immunological release of histamine from leukocytes isolated from blood of 14 allergic, asthmatic patients, was investigated after in vivo or in vitro drug administration. Simultaneously, the effect of this drug on peak expiratory flow rate (PEFR) was estimated. Histamine was assayed by the single isotope-enzymatic method. Obtained results have confirmed that salbutamol is a rather poor inhibitor of histamine release from basophils. Although both administered salbutamol doses (0.5 or 1 mg, i.v.) significantly increased PEFR (p less than 0.01), inhibitory effect on histamine release by allergen was seen only after higher dose administration in vivo (p less than 0.01 against control). Similarly, a small but significant decrease in histamine release by both concanavalin A (by 20%, p less than 0.01) and allergen (by 30%, p less than 0.025) was seen in vitro after incubation of leukocytes with 8 x 10(-6) mol/l salbutamol, whereas 4 x 10(-7) mol/l salbutamol was without effect.

Peroxidation-induced changes of histamine metabolism and transport of its precursor histidine in rat brain synaptosomes.

The metabolism of histamine and transport of its precursor histidine were investigated in rat brain synaptosomes which underwent the ADP-Fe/ascorbate-induced peroxidation. Peroxidation impaired histidine uptake by 40%, and veratridine induced release of it by 25% of maximal uptake. Simultaneously, marked decrease of synaptosomal histamine (HA) content, to about 30% of control value, was found (p less than 0.01). Activity of the two histamine-metabolizing enzymes, histidine decarboxylase (HD) and histamine N-methyl-transferase (HMT), were drastically lowered, by 40% (p less than 0.02) and 60% of control (p less than 0.05), respectively. Pretreatment of rats with glucocorticoid analog, dexamethasone (DMX), 1 mg/kg of body weight, given twice, 20 and 2 h before decapitation, did not influence significantly the effects invoked by peroxidation on HA levels and the activity of HD and HMT, but impaired histidine transport. These results indicate that iron-dependent peroxidation decreases both neuronal pool of histamine and its turnover, which may affect the function of central nervous system. Short pretreatment with dexamethasone does not seem to influence this effect.

Decarboxylation of branched-chain alpha-ketoacids in hepatocytes from alloxan-diabetic rats. The effect of insulin.

The flux through branched-chain alpha-ketoacid dehydrogenase and the activity of the branched-chain alpha-ketoacid dehydrogenase complex were measured in hepatocytes isolated from fed, starved and alloxan diabetic rats. The highest rate of branched-chain alpha-ketoacid oxidation was found in hepatocytes isolated from starved rats, slightly lower in those from fed rats, and significantly lower in diabetic hepatocytes. The amount of the active form of branched-chain alpha-ketoacid dehydrogenase was only slightly diminished in diabetic hepatocytes, whereas the flux through the dehydrogenase was inversely correlated with the rate of endogenous ketogenesis. The same was observed in hepatocytes isolated from starved rats when branched-chain alpha-ketoacid oxidation was measured in the presence of added oleate. In both cases the diminished flux through the dehydrogenase, restored by a short preincubation of hepatocytes with insulin, was paralleled by a decrease of fatty acid-derived ketogenesis. The significance of these findings is discussed in relation to the role of insulin in branched-chain alpha-ketoacid oxidation in liver of diabetic rats.

[Lipid proteinosis of the pulmonary alveoli--diagnostic difficulties]. / Lipoproteinoza pecherzyków plucnych--trudnosci diagnostyczne.

It is the first polish case of diagnosis of alveolar lipid proteinosis diagnosed during life is described. The diagnosis was based on biochemical analysis and morphological assessment in electron microscope of the fluid obtained in bronchoalveolar large in a patient aged 18 years with disseminated radiological pulmonary changes. Diagnostic difficulties are discussed comparing them with the cases of this disease reported as yet in Poland. The importance of bronchoalveolar lavage for the diagnosis is emphasized.

[The effect of salbutamol on histamine release from basophils in vivo and in vitro]. / Wplyw salbutamolu na uwalnianie histaminy z granulocytów zasadochlonnych krwi obwodowej w warunkach in vivo i in vitro.

In vivo and in vitro basophil histamine release inhibition by salbutamol was investigated in patients with bronchial asthma. The study involved 14 patients in stable period of the disease: FEV1 = 66-84% of the normal values. Histamine release was determined following an incubation of the isolated basophils with concanavalin A (Sigma Co., USA) or specific allergens (dust, grass pollens, mites; Bencard, UK). Histamine was assayed with isotope-enzymatic technique according to Shaff and Beaven with histamine N-methyltransferase. Salbutamol administered intravenously in the dose of 1 mg inhibited allergen-induced histamine release from the basophils isolated from patient's blood within 30 minutes. Salbutamol in the concentration of 8 X 10(-6) M inhibited in vitro histamine release induced by an allergen and concanavalin A.

[A case of lymphoid interstitial pneumonia]. / Przypadek limfoidalnego sródmiazszowego zapalenia pluc?

A case of a 16 year old boy with interstitial lymphomatoid pneumonia coexisting with myocarditis, hepatitis, and splenomegaly is presented. The cause of the above mentioned changes despite a thorough and meticulous diagnostic approach which included microscopical examination of the lung, liver biopsies and bone marrow tap could not be made. Corticosteroid therapy did not bring a permanent improvement in the child clinical state. The boy expired quite unexpectedly. The post-mortem examination also did not provide a final diagnosis.

Effect of in vivo glucocorticoid administration on histamine metabolism in rat brain synaptosomes.

The effect of in vivo administration of acetonide triamcinolone (AT) on histamine (HA) metabolism in synaptosomes, highly purified from rat brain by discontinuous Ficoll-sucrose gradient centrifugation, was investigated. AT decreases ability of synaptosomes to actively accumulate HA precursor, L-[U-14C]histidine, measured after rapid centrifugation of synaptosomes through silicone oil, as well as lowers activity of HA catabolizing enzyme histamine N-methyltransferase (by 57%). In contrast, no changes in the level of HA, assayed by the single isotope enzymatic method, and the activity of HA synthesizing enzyme, histidine decarboxylase, were found. The results indicate that under applied conditions glucocorticoid administration may decrease turnover of neuronal pool of HA without significant effect on HA level.

Hypoxia and ischemia modifies histamine metabolism and transport in brain synaptosomes.

Histamine level (HA), the activities of the HA synthetizing enzyme--histidine decarboxylase (HD) and HA metabolizing enzyme--histamine methyltransferase (HMT) and the uptake and release of histidine and histamine were analyzed in synaptosomal preparations obtained from rats with brain hypoxia and ischemia. Hypoxia produced only non-significant changes in all the parameters studied, whereas ischemia induced increase of both enzyme activities and histidine release, with simultaneous decreased of histidine uptake and HA level. The effect of ischemia appeared to be reversible; the changes retreated within 1 h of resuscitation together with the vital functions of rats.