Muscarinic receptor subtypes in equine tracheal smooth muscle

Selective muscarinic receptor antagonists were used to identify muscarinic receptor subtypes in equine trachealis strips. The M₁ receptor antagonist pirenzepine (10^–7 mol/L to 3 × 10^–5 mol/L) and the M₃ receptor antagonist 4-diphenylacetoxy-N-methylpiperidine (4-DAMP, 10^–9 mol/L to 3 × 10^–7 mol/L³) dose dependently inhibited the contractile responses to electrical field stimulation (EFS) and exogenous acetylcholine (ACh). Schild plots yielded a pA₂ value for pirenzepine vs ACh of 6.75 ± 0.09, which is consistent with the affinity for M₂ or M₃ receptors, and a pA₂ value for 4-DAMP vs ACh of 8.47 ± 0.09, which is in agreement with the affinity for M₃ receptors. The M₂ receptor antagonist gallamine (10^–5 mol/L and 10^–4 mol/L) did not affect the response of trachealis to exogenous ACh and low-frequency EFS (0.1–2 Hz) but decreased the responses to high-frequency EFS (4–16 Hz). These results suggest that the muscarinic receptors mediating contractions induced by ACh in equine tracheal smooth muscle are of the M₃ subtype. The lack of an increase in the response to EFS following gallamine suggests that functional prejunctional inhibitory M₂ receptors are not present on the cholinergic nerves innervating equine tracheal smooth muscle.     

Effects of recombinant canine granulocyte colony-stimulating factor on white blood cell production in clinically normal and neutropenic dogs.

Recombinant canine granulocyte colony-stimulating factor (rcG-CSF) was administered to clinically normal dogs, cyclic-hematopoietic dogs, and dogs undergoing autologous bone marrow transplantation, to determine whether rcG-CSF could be used to stimulate WBC production and function in normal and neutropenic dogs. To the normal dogs, rcG-CSF was administered by SC injection at rates of 1 microgram/kg of body weight, q 12 h; 2 micrograms/kg, q 12 h; or 5 micrograms/kg, q 12 h. A significant dose-dependent increase in the WBC count resulted from the stimulation of bone marrow progenitor cells. The increased WBC count was characterized by mature neutrophilia and monocytosis. Neutrophil myeloperoxidase and phagocytic activity were normal in rcG-CSF-treated normal dogs, demonstrating the production of normal functional neutrophils in response to rcG-CSF treatment. Recombinant canine G-CSF prevented neutropenia and associated clinical signs but did not completely eliminate the cycling of neutrophils in cyclic-hematopoietic dogs when it was administered at rates of 1 microgram/kg, q 12 h, and 2.5 micrograms/kg, q 12 h. The time to bone marrow reconstitution was not decreased in dogs treated with rcG-CSF at a rate of 2.5 micrograms/kg, q 12 h, for 13 days following autologous bone marrow transplantation. On the basis of our findings, we suggest that treatment with rcG-CSF is an effective way to stimulate myelopoiesis in dogs, but that the dose of rcG-CSF required to stimulate WBC production will vary depending on the cause of neutropenia. Recombinant canine G-CSF should be useful in stimulating production and maintaining function of WBC for treatment of clinical diseases seen commonly in veterinary practice.     

Mesangioproliferative Glomerulonephritis in Mink with Encephalitozoonosis

Renal specimens from 6 mink with encephalitozoonosis were studied by light and electron microscopy and immunohistochemistry. The glomeruli of affected kidneys had a mesangioproliferative glomerulonephritis which was characterized by an increase in mesangial cells and matrix in most glomeruli. Some glomeruli were partially or completely sclerosed. There were protein or granular casts in the cortical and medullary tubules. Interstitial nephritis, vasculitis and tubular cysts were found. Electron microscopy demonstrated extensive matrix and increased cellularity in the mesangial areas. Glomeruli showed segmentally thickened or wrinkled capillary basement membranes. Electron dense deposits were found in the glomerular basement membranes and mesangium. Peroxidase-anti-peroxidase immunohistochemistry demonstrated that IgG and IgM positive material was present as granular deposits in the glomerular basement membrane and occasionally in the mesangium.     

Potential effects of glucocorticoids on serum iron concentration in dogs

Prednisone was give norally(2mg/kg b.i.d.) to seven healthy mixed breed dogs for 3 consecutive days. Serum iron concentration increased significantly (p < 0.05) from 142 +/- 26 micro g/dl (mean +/- SE) before a drug adminis- tration on Day 0 to a maximum of 307 +/- 47 micro g/dl on Day 2, and returned to the Day 0 value by Day 5. Mean total iron binding capacity did not vary more than 25% from the Day 0 value during the 9 day long study. The percent saturation of transferrin with iron increased from 33 +/- 6% on Day 0 to a maximum of 71 +/- 9% on Day 3. This determination had decreased to 34 +/- 3% on Day 5. No statistically significant changes occurred in these parameters studied in six control dogs that were not given the drug. To determine whether serum iron concentration might be correlated with endogenous serum cortisol concentration, these tests were determined in serum collected from nine dogs at 7 a.m., 3 p.m., and 11 p.m. each day for 3 consecutive days. Serum iron concentration was lower at 7 a.m. (147 +/- 9 micro g/dl) than at 3 p.m. (164 +/- 9 micro g/dl) or 11 p.m. (159 +/- 10 micro g/dl). Likewise serum cortisol was lower at 7 a.m. (1.29 +/- 0.18 micro g/dl) than at 3 p.m. (1.49 +/- 0.19 micro g/dl) or 11 p.m. (1.51 +/- 0.22 micro g/dl). There was a significant positive linear correlation between serum iron and serum cortisol concentrations when they were compared using mean values for each dog. From these studies, it appears that exogenously administered glucocorticoids and endogenous increases in serum cortisol concentrations may result in increased serum iron concentrations in dogs.     

Haemagglutination-inhibiting antibodies against arboviruses of the families Togaviridae and Bunyaviridae in birds caught in southern Moravia, Czechoslovakia

A total of 295 birds belonging to 19 species of 7 families of wild Passeriformes were examined by haemagglutination-inhibition test. The birds were caught for an international research program "Balt" at the time of autumn migration (August-September 1984). Their blood sera were examined for antibodies against 6 arbovirus antigens of the genera Alphavirus (Sindbis-SIN) and Flavivirus (tick-borne encephalitis-TBE, West Nile-WN) and family Bunyaviridae (Tahyna-TAH, Calovo-CVO and Bhanja-BHA). Antibodies against all studied viruses were detected at different frequencies: SIN 6.4%, TBE 7.1%, WN 9.7%, TAH 16.3%, CVO 12.1%, and BHA 1.0%.     

Some diagnostic, biologic and morphologic characteristics of Francisella tularensis strains isolated from the ticks Ixodes ricinus (L.) in the Prague agglomeration

Four infectious agents were isolated from the ticks Ixodes ricinus (L.) collected in the recreational area and park-forest of Prague. On the basis of cultivation, staining, biochemical, serologic properties, pathogenicity for animals and histological tests they were identified as Francisella tularensis with the following features: they are short, gram-negative rods of approximate dimensions of 0.3 X 0.8 micron, growing in enriched media after 3-4 day incubation at 37 degrees C. They form small circular, at first transparent, later greyish turbid colonies with regular rims. They are little active biochemically. They are susceptible to streptomycin and some broad spectrum antibiotics. They react positively with tularemic serum, but in lower titres than those in which this serum reacts with standard antigen. The microbes are highly pathogenic for mice, guinea-pigs, young rats, in which a massive bacteriemia occurs before death, but they do not kill rabbits. They multiply well in chick embryo, but do not grow in cell or tissue cultures. The most important histologic changes were observed in liver and spleen of mice. No pathologic changes were found in brain, lungs, heart, kidneys. Necroses were found in liver and in their marginal zones the microbes were present. Conspicuous were changes in numerous hepatocytes which became enlarged due to microbial multiplication and finally transformed into "sacs" packed with microbes. Histological and electronoptical examination showed that these are intracellular parasites fringed with a light lytic zone. Discussed is the problem to what extent the properties of the isolated strains are typical of F. tularensis as well as the importance of their detection from the aspect of epidemiology and differential diagnostics.     

Further characterization of dandelion yellow mosaic virus from lettuce and dandelion

A damaging virus isolated in the Netherlands from lettuce was studied and compared with a virus isolated from dandelion orginating from Czechoslovakia. It was found to biologically resemble dandelion yellow mosaic virus incompletely described from dandelion and lettuce in Great Britain (Kassanis, 1944, 1947) and from dandelion in Germany (Hein, 1963). Mechanical transmission was greatly improved by buffer solution and transmission byMyzus persicae seemed to be in the non-persistent manner. Longevity in vitro of the virus hardly exceeded one day. Thermal inactivation was between 60 and 65 °C and the dilution end-point was between 10 000 and 100 000. It was still infectious in leaf material dried and stored over CaCl₂ at 4 °C for 6 1/2 years. The virus was isolated and purified with difficulty and was found to consist of one type of spherical particle of ca 30 nm diameter, with a sedimentation coefficient of 159 S, a buoyant density of 1.42 g.cm^?3 and an A₂₆₀/A₂₈₀ ratio of 1.67. An antiserum was prepared with a titre of 256 in the agar double-diffusion test. The virus could be identified in crude extracts from lettuce andChenopodium amaranticolor by enzyme-linked immunosorbent assay (ELISA), but not by agar double diffusion. It could only be visualized in crude sap in the electron microscope after trapping of virus particles on antiserum-coated grids. The virus cannot yet be assigned to any known virus group. It is of potential economic importance to lettuce because of its occurrence in widely differing regions in Europe, its aggressiveness and virulence on 22 out of 23 lettuce cultivars tested (and on endive) and its pathogenicity toLactuca genotypes which are resistant to lettuce mosaic virus and other important pathogens of lettuce. ‘Laibacher Eis’ was the only cultivar showing some tolerance.