Cellular localization of bovine pancreatic trypsin inhibitor and related molecular forms in bovine lung

Summary In addition to bovine pancreatic trypsin inhibitor (BPTI), three BPTI-related molecular forms (isoinhibitors I, II and III) were isolated from bovine lung by affinity chromatography on immobilized trypsin and subsequently purified by Fast Protein Liquid Chromatography. These inhibitors are identical to the isoinhibitors previously isolated from bovine spleen. Their localization in bovine lung was studied by immunohistochemical techniques, using two different immunoglobulin preparations, selectively recognizing BPTI or the other molecular forms.BPTI-related immunoreactivity was found to be restricted to isolated cells, often identified as mast cells by Toluidine Blue staining. In contrast, isoinhibitor-related immunoreactivity, which also occurs in the mast cells, is present in a number of other cell types. These types include: (i) the smooth muscle cells of different calibre vessels, (ii) the ciliated cells of the bronchial epithelium and the related mucus, and (iii) many cells at alveolar level.Comparison of these data with previous results obtained for bovine spleen suggest multiple physiological roles for these inhibitors.     

Bidirectional effect of met-enkephalin on macrophage effector functions

Summary Met-enkephalin (ME) exerts a bimodal effect on functional activities of rat peritoneal macrophages (PM); in a range of low concentration (10^-9-10^-7 M) antibody dependent cellular cytotoxicity (ADCC)was markedly stimulated with a simultaneous decrease of Fc receptor (FcR) mediated phagocytosis while the opposite was observed at 10^-6-10^-5 M concentrations.Studying the possible underlying mechanism(s) the followings were recorded: (1) ME in all applied concentrations induced an early Na⁺ influx which was followed by a Ca²⁺ efflux in the range of low concentrations. In the range of high concentrations Na⁺ influx was accompanied by a Ca²⁺ influx. (2) ME at 10^-8 M concentration induced a rise in cGMP level with a plateau in the 60–120th min of incubation. This effect was prevented by 10^-5 M of naloxone. At 10^-6 M concentration a transient rise of cAMP level was recorded which was not affected by naloxone. (3) Verapamil in 10^-6 M abolished both the Ca²⁺ influx and the rise in cAMP level induced by 10^-6-10^-5 M ME but not the rise in cGMP level induced by lower ME concentrations. (4) cAMP elevation by high ME concentrations was abolished by enkephalinase inhibitory puromycin. (5) PM-enkephalinase as assessed by the cleavage of fluorogenic substrate L-alanine beta naphthylamide (ABNA), was inhibited by 10^-6-10^-5 M of ME. This inhibition was abolished by verapamil, but not affected by naloxone. In the range of low concentrations ME appears to act on specific delta opioid receptors and its action is positively coupled to guanylate cyclase. In relatively higher concentrations ME-action is not mediated by specific delta opioid receptors and it appears to involve Ca²⁺ influx, adenylate cyclase activation as well as the processing of hormone by PM-enkephalinase.     

Aerodynamics of Ephedra trifurca

Computer simulations are used to predict the behavior of pollen grains with different physical properties within the acceleration field created around the ovules of the gymnosperm Ephedra trifurca. A modelling procedure is given that (1) calculates the number of pollen grains captured by an ovule's pollination-droplet and (2) gives a correlation between pollination efficiency and the physical properties (= mass, size) of different types of pollen. Based on this procedure, the number of Ephedra pollen grains captured by micropyles can be less than the number captured from other species. However, the mass and size of Ephedra pollen grains appear to coincide with those predicted to yield a local maximum of pollination efficiency, i.e. slightly larger or smaller values of either mass or size would decrease the probability of capture. In addition, the properties of Ephedra pollen grains operate synergistically in the aerodynamic environment around ovules and are focused to collide with pollination-droplets. By analogy, the properties of Ephedra pollen coincide with those predicted for a localized adaptive peak. The physical properties of pollen grain types other than E. trifurca that can maximize pollen capture are not generally represented in the aerobiology of Ephedra during the pollination season. Therefore, the phenology of pollen release, community taxonomic-composition, and the physics of particle capture play collectively important roles in the reproductive success of Ephedra trifurca.     

Monoclonal antibodies modify acetylcholine-induced ionic channel properties in cultured chick myoballs

Summary Monoclonal antibodies directed against the cholinergic binding site of the acetylcholine receptor were found to alter the ion channel properties in cultured chick myoballs. Time and dose dependent reduction in acetylcholine sensitivity was observed. Noise analysis experiments indicated a decrease in the mean single channel conductance and an increase in the mean single channel open time.     

Deformation of the Diastolic Left Ventricle: I. Nonlinear Elastic Effects

A linear incremental finite element model is used to analyze the mechanical behavior of the left ventricle. The ventricle is treated as a heterogeneous, non-linearly elastic, isotropic, thick-walled solid of revolution. A new triaxial constitutive relation for the myocardium is presented which exhibits the observed exponential length-passive tension behavior of left ventricular papillary muscle in the limit of uniaxial tension. This triaxial relation contains three parameters: (a) a “small strain” Young's modulus, (b) a Poisson's ratio, and (c) a parameter which characterizes the nonlinear aspect of the elastic behavior of heart muscle. The inner third and outer two-thirds of the ventricular wall are assumed to have small strain Young's moduli of 30 and 60 g/cm², respectively. The Poisson's ratio is assumed to be equal to 0.49 throughout the ventricular wall. In general, the results of this study indicate that while a linearly elastic model for the ventricle may be adequate in terms of predicting pressure-volume relationships, a linear model may have serious limitations with regard to predicting fiber elongation within the ventricular wall. For example, volumes and midwall equatorial circumferential strains predicted by the linear and nonlinear models considered in this study differ by approximately 20 and 90%, respectively, at a transmural pressure of 12 cm H₂O.     

DNA damage induced by phorbol ester-stimulated neutrophils is augmented by extracellular cofactors. Role of histidine and metals

Activated neutrophils cause extensive DNA damage in neighboring nonphagocytic cells. To determine whether compounds in the extracellular milieu participate in the DNA damage process, murine neutrophils were cocultivated with target tumor cells in media of varying composition. Using the alkaline elution assay, it was found that the level of strand breaks induced was significantly higher (2.8-fold) in complex cell culture media than in minimal phosphate-buffered saline. Addition of amino acids in general and of histidine in particular increased the level of damage nearly to that observed in complete media (2.7- and 2.1-fold, respectively). The histidine stimulation was concentration-dependent and reached a maximum at 100-400 microM. The mechanism whereby this occurred is not proven but probably derived from chelation of metals and participation in a site-specific Fenton reaction. Addition of the cell-impermeable chelator EDTA dramatically inhibited induction of strand breaks by neutrophils in complete media and prevented the enhancement of damage induced by histidine in phosphate-buffered saline. None of the effects on neutrophil-induced damage could be attributed to modulation of the oxidative burst activity of the cells (O2- and H2O2 production). Histidine also enhanced induction of strand breaks by reagent H2O2. However, EDTA had no effect or actually increased the level of damage induced by both a bolus of H2O2 and a flux of H2O2 generated by glucose oxidase. The cell-permeable chelator o-phenanthroline inhibited both neutrophil- and H2O2-induced damage. The results indicate that secondary reactions involving extracellular amino acids and metals contribute significantly to neutrophil-induced DNA damage to neighboring cells. Moreover, the data show that the mechanism whereby neutrophils induce this damage cannot be attributed solely to secretion of H2O2.     

Cytogenetic characteristics of a population of Chironomus riparius Meigen 1804 (Diptera, Chironomidae) from a polluted Po river station

A population of Chironomus riparius from a Po river station near Moncalieri (a trace-metal polluted station) was studied. In this population was established a great variability of band structure of polytene chromosomes as well as paracentric heterozygous inversions, deletions, deficiencies, partial breaks, diploid chromosome fragments, and changes in functional activity and appearance of heterochromatin. In arms A through F, some bands had an increased size compared to the standard chromosomic map. Some bands appeared in a heterozygous or normal homozygous state or were amplified. In all arms, many condensed stable bands appeared in the decondensed state when compared to the standard map. Asynaptic zones in arms E and G as well as heterozygous Balbiani rings and NORs were established. Very often the 4th chromosome was almost completely heteropycnotic and looded like a pompon chromosome. For the first time in this species, a high frequency of ectopic pairings of different arms was observed. Telomeric regions involved in ectopic pairings had a granular appearance, as did some centromeres. The hypothesis is advanced that such a high frequency of structural rearrangements could be correlated with genomic distribution of specific mobile elements.     

An artificial bifunctional enzyme, γ-glutamyl kinase/γ-glutamyl phosphate reductase, improves NaCl tolerance when expressed in Escherichia coli

Summary An artificial bifunctional enzyme, -glutamyl kinase/-glutamyl phosphate reductase, was obtained by fusing the Escherichia coli genes proA and proB. The proB gene was fused to the 5-end of the proA gene with a linker encoding five amino acids. When expressed in E. coli enhanced intracellular concentrations of proline were observed. At 0.6 M NaCl the growth rates for the strain carrying the fusion enzyme and a control harbouring a plasmid encoding the wild-type enzymes were 320 and 530 min, respectively.     

The PARP and rRNA promoters of Trypanosoma brucei are composed of dissimilar sequence elements that are functionally interchangeable.

The African trypanosomes express two major surface proteins, the variant surface glycoprotein (VSG) and the procyclic acidic repetitive protein (PARP). The RNA polymerase that transcribes the VSG and PARP genes shares many characteristics with RNA polymerase I. We show that although there is very little similarity in nucleotide sequence, the functional structure of a trypanosome rRNA promoter is almost identical to that of the PARP promoter. Further, domains from the PARP promoter can functionally substitute for the corresponding parts of the rRNA promoter, and vice versa.