Use of monoclonal antibodies against human HLA II antigens for the detection of bovine B lymphocytes and macrophages]

The crossreactivity of mouse monoclonal antibodies (MoAbs) (Tab. I) prepared against human HLA-DR and HLA-DP antigens was studied in various bovine cells: lymphocytes from lymph nodes and peripheral blood, adherent (B) and nonadherent (T) lymphocytes, monocytes, granulocytes and platelets. In the immunofluorescence test, MoAbs Bra13, Bra14, Bra20, Bra22, Bra30, Bra70, HL-38 reacted with bovine B lymphocytes and monocytes, but not with other tested cells (Tab. III, IV). These antibodies, except Bra22, were positive with B lymphocytes in the complement dependent cytotoxic test (Tab. II). The similarity of the bovine antigens and HLA-DR antigens determined by used MoAbs was also proved by immunoblotting. Monoclonal antibodies Bra38 and BraFB6 did not react with the bovine cells and separated antigens. The epitope (HLA-DR) recognized by the antibody Bra38 is probably absent in cattle. The presence of HLA-DP analogue determined by the antibody BraFB6 has not been confirmed. The crossreactive MoAbs could be used for the detection of B lymphocytes and macrophages in veterinary immunology.     

Regulation of Capacitation of Canine Spermatozoa during Co-culture with Heterologous Oviductal Epithelial Cells

Progress of essential steps of the capacitation is coordinated in the oviductal isthmus, where sperm are stored in close contact with the epithelium. A crucial capacitational event is the phosphorylation of sperm membrane proteins. Regulation of the tyrosine phosphorylation by the oviduct has not been examined in dog sperm yet. The aim of this work was to study the effect of dog sperm binding to porcine oviductal epithelium on capacitation‐induced cellular and molecular changes. Epithelial cells were stripped from the oviducts of post‐puberal sows and cultured for 5–7 days at 39°C and 5% CO₂ on Biomatrix‐covered Chamber slides. Sperm washed through Percoll was co‐incubated with the oviductal epithelium cell cultures in a bicarbonate Tyrode's medium. During co‐incubation, sperm membrane changes, the state of tyrosine phosphorylation and motility were determined after 3, 30, 90, 180, 240 and 360 min. Significant increases in the percentage of capacitated and dead cells were observed in unbound sperm, while bound sperm remained uncapacitated, live and motile. An increasing tyrosine phosphorylation of tail proteins in bound, unbound and control sperm suspensions and a subsequent phosphorylation of head proteins in unbound and control sperm suspensions were observed. A significant difference regarding head phosphorylation (p < 0.05) was found between sperm bound to oviductal epithelium and unbound sperm. Binding occurred mainly in sperm with non‐ phosphorylated heads, while higher proportions of phosphorylated cells were found in unbound populations. The head phosphorylation progressed significantly during incubation in unbound spermatozoa (p < 0.05); however, it was suppressed in population of sperm attached to oviductal epithelium. Significant correlations between motility parameters related to hyperactivation and tail phosphorylation were found in unbound sperm. These observations support the hypothesis that spermatozoa with non‐phosphorylated heads preferentially attach to epithelial cells. It can be concluded that tyrosine phosphorylation of head membrane proteins and capacitation are delayed in canine spermatozoa being in closed contact with oviductal epithelium.     

Apparent independence of glucose tolerance and body fat content in 8‐week‐old broilers

1. A possible relationship between glucose tolerance and body‐fat content was examined in broilers selected at 2 and 4 weeks of age for fast or slow glucose disposal.

2. At 8 weeks of age, selected chickens were different in glucose tolerance but similar in body weight, food conversion efficiency, carcass composition and glucose‐induced insulin release.

3. Therefore, variations in glucose regulation and insulin sensitivity which are detectable at an early age, do not appear to be related to body composition in 8‐week‐old broilers.     

Effects of atmospheric ammonia on pulmonary bacterial clearance in the young pig.

Young pigs were exposed to an aerosol of a nonpathogenic strain of Escherichia coli and then were retained in air-pollutant exposure chambers for a 2-hour clearance period. In series 1 (n = 80 pigs), 40 exposed young pigs (principals; 15.5 days of age) were placed in an atmosphere of filtered room air + 50 ppm of atmospheric NH3 during the clearance period; control pigs were exposed to filtered room air without added NH3. In series 2 (n = 24 pigs), 12 exposed young pigs (principals; 6.2 days of age) were similarly maintained, but at a lower concentration of atmospheric NH3 (75 ppm). At the end of the clearance period pigs were killed and pulmonary bacterial clearance was determined. Pigs kept in the NH3-contaminated atmospheres (either concentration) harbored more bacteria, on the average, in their lungs than did the controls. If series 1 and 2 data were combined, pigs kept in the NH3-contaminated atmospheres had 51% more bacteria in their lungs than did the controls. Pulmonic weight and ratio of pulmonic weight to body weight of pigs kept in the NH3-contaminated atmosphere were greater than those of the controls in series 1, but not in series 2. Gross and histopathologic examinations of lung tissue generally revealed no differences between controls and principals in either series 1 or 2.     

Studies on methods for the determination of amino acid requirements for metabolic balance based on the oxidation rate of 14C labeled lysine to 14CO2]

Male experimental rats (100 gm liveweight) were distributed into 10 groups of 8 animals each and received balanced diets, with the exception of lysine which was added to the diets in graded amounts in such a way that the lysine content of the diets ranged from 2.44 to 5.92 gm/16 gm N. After a feeding period of 7 days the animals received 3H- and 14C lysine injected intraperitoneally, 4 animals of each group were investigated for the total CO2 excretion and 14CO2 excretion during the first 2 hrs after the injection and for the urinary excretion of radioactivity (48 hours). The remaining animals in each group were used for determining the plasma amino acids and for establishing the specific radioactivity of free lysine in the liver and muscles after an 1-hour incorporation period. Total CO2 excretion was not found to be influenced by the lysine contents while the level of excretion of 14C activity through CO2 and that of specific 14C activity of CO2 increased with increasing lysine concentrations. This produced a broken curve pattern, showing an increased release of 14CO2 (under maintenance conditions) if the diet contained 4 gm lysine/16 gm N and more. Investigations for the specific 14C activity of free lysine in the liver, the main site of lysine oxidation, showed that the increase in 14CO2 release was due to an enhanced rate of lysine catabolism and was not brought about by changes in the pool volume or in specific radioactivity. The levels of urinary 14C excretion were not found to be related to the lysine content of the diets, whereas the curve pattern of 3H excretion observed 5 to 8 hrs after injection was similar to that of 14CO2 excretion. The lysine content of blood plasma and the content of free lysine in the liver increased continuously with increasing levels of dietary lysine. The methodological studies made in the present paper showed that in scientific research a determination of amino acid requirements on the basis of CO2 oxidation data may be a very exact and sensitive method. It will also yield values for maintenance requirements.     

Littoral spawning habitats of three southern Arctic charr (Salvelinus alpinus L.) populations

Abstract Arctic charr populations in southern latitudes are nonmigratory, with all life‐stages limited to freshwater lakes and in‐ or out‐flowing tributaries. Although many of these populations are reported to also spawn in lake littorals, little is known about the physical characteristics of putative spawning grounds. A total of 23 discrete spawning sites within three Irish lakes were located by fyke netting of spawning adults and snorkelling in littoral habitats. Spawning sites were found to be long, narrow strips running parallel to the shore at a maximum depth of 124 cm. Spawning sites were limited to areas of coarse mineral substrate with an adequate (c. 8 cm) depth of clean interstitial spaces. In individual lakes, combined areas of spawning sites made up 0.4–0.7% of available littoral. Egg densities varied considerably between sites (33–900·eggs m^?2) and were significantly correlated with gradient and width of spawning sites. No evidence of redd digging was found. The shallow, localised and restricted nature of spawning grounds makes such populations vulnerable to anthropogenically induced postoviposition changes in surface water level, eutrophication processes such as increased lake sedimentation and elevated nutrient status.     

The thermal antinociceptive effects of a high-concentration formulation of buprenorphine alone or followed by hydromorphone in conscious cats

ObjectiveTo evaluate the thermal antinociceptive effects of a high-concentration formulation of buprenorphine alone or followed by hydromorphone in conscious cats.Study designRandomized, blinded, placebo-controlled crossover study design.AnimalsA total of six purpose-bred, adult female ovariohysterectomized Domestic Short Hair cats.MethodsCats were allocated into three treatments each consisting of two injections, subcutaneous then intravenous (IV) administration, 2 hours apart: treatment SS, two injections of 0.9% saline; treatment BS, buprenorphine (0.24 mg kg^–1, 1.8 mg mL^–1) and saline; and treatment BH, buprenorphine (0.24 mg kg^–1) and hydromorphone (0.1 mg kg^–1). Skin temperature (ST) and thermal threshold (TT) were recorded before (baseline) and for 24 hours following first injection. TT data were analyzed using mixed linear models and a Benjamini–Hochberg sequential adjustment procedure (p < 0.05).ResultsThere were no significant differences among treatments for baseline ST and TT values, treatment SS over time and between treatments BS and BH. Compared with baseline, TT was significantly increased at all time points in treatments BH and BS except at 2 hours in treatment BS. TT was significantly higher than SS at 3–18 hours and 4–12 hours for treatments BS and BH, respectively. Maximal increases in TT were 47.5 °C at 2 hours, 53.9 °C at 3 hours and 52.4 °C at 6 hours in treatments SS, BS and BH, respectively.Conclusions and clinical relevanceAdministration of IV hydromorphone following high-concentration buprenorphine provided no additional antinociception and decreased the duration of effect when compared with high-concentration buprenorphine alone. Alternative analgesics should be considered if additional analgesia is required after administration of high-concentration buprenorphine.     

TRUNCATION ARTIFACT IN MAGNETIC RESONANCE IMAGES OF THE CANINE SPINAL CORD

The truncation artifact in magnetic resonance (MR) images is a line of abnormal signal intensity that occurs parallel to an interface between tissues of markedly different signal intensity. In order to demonstrate the truncation artifact in sagittal images of the canine spinal cord and the effect of changing spatial resolution, we conducted an experimental in vitro study. A section of fixed canine spinal cord was imaged using a 1.5T magnet. Spatial resolution was increased by increasing the acquisition matrix and reconstruction matrix, producing series of T2‐weighted (T2w) images with the following pixel sizes: A, 1.6 (vertical) × 2.2 mm² (horizontal); B, 1.2 × 1.7 mm²; C, 0.8 × 1.1 mm²; D, 0.4 × 0. 6 mm². Plots of mean pixel value across the cord showed variations in signal intensity compatible with truncation artifact, which appeared as a single, wide central hyperintense zone in low‐resolution images and as multiple narrower zones in high spatial resolution images. Even in images obtained using the highest spatial resolution available for the MR system, the edge of the spinal cord was not accurately defined and the central canal was not visible. The experiment was repeated using an unfixed spinal cord specimen with focal compression applied to mimic a pathologic lesion. Slight hyperintensity was observed within the spinal cord at the site of compression although the cord was normal histologically. Results of this study suggest that caution should be applied when interpreting hyperintensity affecting the spinal cord in T2w sagittal images of clinical patients because of the possibility that the abnormal signal could represent a truncation artifact.