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.     

An evaluation of the Olsen test as a measure of plant-available phosphorus in grassland soils derived from basalt parent material

Abstract. Anecdotal and circumstantial evidence have suggested that the Olsen test underestimates plant-available phosphorus (P) in basaltic soils in Northern Ireland. Therefore, the ability of this test to predict plant-available P in basaltic (and non-basaltic) soils was investigated by regressing Olsen-P data against herbage P indices calculated from plant tissue test data using the diagnosis and recommendation integrated system. The average Olsen-P concentration for a range of fields situated on basaltic soils was considerably lower than the average Olsen-P concentration for a range of fields situated on non-basaltic soils, and yet mean sward P status, as given by the herbage P indices, was similar for both groups of fields. Herbage P indices were also much better correlated with Olsen-P measurements in non-basaltic soils than in basaltic soils. Furthermore, at low Olsen-P values (≶9mgPL⁻¹) some swards on basaltic soils were genuinely deficient in P, while others were sufficient or even in surplus for this nutrient. The results confirm that Olsen-P is inadequate as a predictor of plant-available P in basaltic soils. It is concluded that an alternative soil test is needed to provide a reliable assessment of plant-available P in basaltic soils, to prevent overuse of fertilizer and manure P and to minimize the amounts of P entering local watercourses.     

Some Comments on Gerechte Designs

In a companion paper we derived the correct analysis for gerechte designs with uncorrected errors. Here we show that this correct analysis cannot be justified by the usual randomization argument. However, when the regions are rectangular there is a randomization procedure which validates an analysis with three separate error terms. We also outline other developments in design and analysis that may be more satisfactory.     

Outcome of limb amputations in wapiti: 13 cases (1995-2001).

The object of this investigation was to determine the outcome of limb amputation in wapiti. Medical records of 13 wapiti that underwent limb amputation were reviewed to determine age, weight, sex, injury preceding amputation, limb amputated, amputation location, length of hospital stay, complications, and outcome. Ages ranged from 3 months to 8 years and weights from 70 kg to 280 kg. All animals were female. Eleven animals sustained catastrophic long bone fractures prior to amputation. Five animals had front limb amputations and 8 had hind limb amputations. Of these animals, 1 with a front limb amputaiton and 5 with hind limb amputations survived. Postsurgical inability to stand is associated with hospitalization of > 1 day. Of the 13 wapiti reviewed, 5 with hind limb amputations have produced and reared offspring. Hind limb amputation can be considered a viable alternative to euthanasia when catastrophic orthopedic injuries occur in female wapiti.     

MAGNETIC RESONANCE IMAGING OF CEREBRAL CORTICAL NECROSIS (POLIOENCEPHALOMALACIA) IN A DOG

A 3-year-old neutered female mixed breed dog was examined because of severe, generalized seizure activity, tetraparesis, and encephalopathic signs. Cerebrospinal fluid (CSF) evaluation was unremarkable except for a mild increase in protein. Serum and CSF titers for infectious diseases were negative. Magnetic resonance (MR) imaging examination of the brain was performed and lesions were found within the cerebral gray matter of the temporal and parietal lobes. The lesions had increased signal intensity on T1, T2, and proton density-weighted images. There was mild inhomogeneous enhancement following intravenous contrast medium administration. Neurologic status improved and the seizures were well controlled, but the dog never regained normal mentation and euthanasia was performed 10 weeks after initial evaluation. At necropsy, severe cerebral cortical necrosis was found in the regions corresponding to the lesions seen on MR imaging examination. Large numbers of fat-containing macrophages (gitter cells) were found within these areas, and are thought to be responsible for the characteristic hyperintensity seen on the MR images.     

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.     

Herniation of small intestine through the right lateral ligament of the bladder in a bull

A three year old Charolais bull was examined because of anorexia and depression of five days duration and progressive abdominal distention. Intestinal obstruction was diagnosed. A right flank celiotomy allowed diagnosis and correction of a herniated jejunoileal loop through the right lateral ligament of the bladder. An annular constriction located in mid-ileum was by-passed by ileocecal side-to-side anastomosis. The bull's appetite and fecal production returned to normal within 72 hours of surgery. Ten months following discharge the bull was reported to be well.     

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.