Ultrastructure of bovine fetal thymocytes forming non-immune spontaneous erythrocyte rosettes

The ultrastructural appearance of bovine fetal thymocytes forming non-immune spontaneous erythrocyte rosettes was studied. The thymocytes-forming rosettes were found to be small, intermitotic lymphocytes. The attachment between the thymocytes and the erythrocytes were usually observed as small contact points.     

Immune function in dairy cows related to energy balance and metabolic status in early lactation

Two feeding experiments were carried out in 2 successive years with 28 cows of the Norwegian Red Cattle (NRF) in each experiment. The cows were randomly distributed into 4 groups and subjected to different feeding regimens from 1 month prior to calving until 12 weeks after calving. The experimental design was factorial (2 x 2) with respect to protein content of the concentrate (17.5% digestible crude protein (DCP) v.s. 12.5% DCP) and concentrate allowances (standard v.s. substandard allowances after calving). Silage was offered ad libitum. Samples for estimation of serum immunoglobulin-G, white blood cells and lymphocyte responses to the mitogens phytohemagglutinin, concanavalin A and pokeweed mitogen were collected 4 weeks prior to expected calving, and 2, 4 and 8 weeks after calving. The levels of milk immunoglobulin-G were estimated at calving and 2, 4 and 8 weeks after calving. A significant positive relationship was found between the estimated energy balance and the lymphocyte response to mitogens. Little evidence was found for the existence of a significant relationship between the immunologic parameters and plasma indicators of metabolic status. The lymphocyte response to phytohemagglutinin and levels of serum immunoglobulin-G increased, while levels of milk immunoglobulin-G decreased during the period from calving to 8 weeks after calving. Increased milk somatic cell counts were associated with a significant decrease in the lymphocyte responses to mitogens.     

Effect of Date of Planting, Genotype and Source of Seed on Disease Incidence, Yield, and Agronomic Performance of Soybean

Four soybean genotypes chosen from four seed sources were planted in a factorial arrangement on four dates in a split-plot design at the Alabama A & M Experimental Station on a Decatur silt clay loam soil. The highest vigor index was recorded from seeds harvested from the 4th date of planting. These seeds had the lowest level of Diaporthe phaseolorum var. caulivora infection, indicating that vigor index, as a measurement of seed quality, was negatively correlated with the level of infection by soybean stem canker pathogen. Seed yield was highest at the 2nd date of planting and lowest at the 4th date of planting. Tracy-M and Bedford confirmed previous reports on their resistance and susceptibility, respectively, to soybean stem canker. All the genotypes performed their best at the 2nd date of planting. Source of seeds appeared to have little effect on the performance of the plant in the field.     

The Effect of Storage on Ammonia Concentration in Canine Packed Red Blood Cells.

Objective: To determine the effect of storage on ammonia concentration in canine packed red blood cell (pRBC) units.
Design: In vitro and in vivo study.
Setting: University Veterinary Teaching Hospital.
Interventions: Ammonia concentration was measured in 7 units of canine pRBC prepared in citrate-phosphate-dextrose (CPD) and Adsol^a on Days 1 and 35 of storage. Ammonia was measured in 4 additional units of canine pRBC on Days 0, 7, 14, 21, 28, and 35. Plasma ammonia was also determined in 5 anemic dogs receiving pRBC.
Measurements and Main Results: Ammonia concentration increased from 73 ± 15 mmol/L (mean ± SD) on Day 1 to 800 ± 275 mmpl/L on Day (p<0.001). When measured every 7 days in 4 units of canine pRBC, ammonia concentration increased from 23 ± 8 mmol/L on Day 0 to 179 ± 13 mmol/L (Day 7), 276 ± 56 mmol/L (Day 14). 383 ± 47 mmol/L (Day21), 466 ± 30 mmol/L (Day 28), and 562 ± 27 mmol/L (Day 35) (p<0.05 for all comparisons). In a preliminary study, plasma ammonia concentration measured in blood samples from 5 anemic dogs without primary liver disease immediately before and after transfusion with 5–10 ml/kg of stored pRBC remained in the normal reference range.
Conclusions: The ammonia concentration in stored canine pRBC increased markedly with time. In this preliminary study, ammonia concentrations in dogs without primary liver disease did not increase above the reference range after transfusion with pRBC.     

EFFECT OF DEPENDENCY VERSUS NONDEPENDENCY ON LUNG LESION VISUALIZATION

Paraffin blocks and mineral oil were used as a model to determine the effect of dependency versus nondependency on radiographic visualization of lung lesions in lateral thoracic radiographs. It was concluded that the increased opacity of the material surrounding the lesion, not contact between the heart and the lesion, was responsible for the inability to detect lung disease in the dependent lung. The results were tested in dogs with pneumonia in the right middle lung lobe. When the dog was in right lateral recumbency, the dependent right lung was increased in opacity and decreased in volume and the pulmonary lesion was difficult to detect. When the dog was in left lateral recumbency, the nondependent right lung was increased in volume and decreased in opacity and the pulmonary disease was clearly visible. A single recumbent lateral radiograph must not be used to assess a dog with suspected lung disease because lesions in the dependent lung lobes may not be detected.     

The growth of swine bone marrow cells in the presence of heterologous colony stimulating factor: Characterization of the developing cell population

Conditioned medium prepared from the mouse fibroblast cell line L 929 which is known to produce colony-stimulating factor active on mouse bone marrow cells was also able to stimulate the growth of swine bone marrow cells in a liquid culture system. During the first 4 days of culture mononuclear phagocyte and granulocyte colonies developed. Prolonged cultured cells were classified belonging to the macrophage lineage by morphological and cytochemical criteria. These cells fulfill also functional characteristics for macrophages, like expression of Fc receptors, immune phagocytosis and production of prostaglandins. These bone marrow-derived macrophages could also be activated with LPS and lymphocyte-derived mediators.     

Demonstration of inflammatory cell population changes in rat lungs in response to intratracheal instillation of spring wheat dust using lung enzymatic digestion and centrifugal elutriation

The inhalation of grain dust by grain workers is responsible for a large number of pulmonary pathophysiologies. These problems may be acute or chronic and may be mediated by the chronic activation of the immune system. Constant inflammatory states in the lung may eventually lead to tissue damage and respiratory deficit. This study was designed to measure the changes in the relative number of inflammatory cells in peripheral blood, bronchoalveolar spaces, and lung interstitium that occur in response to intratracheally instilled airborne spring wheat dust in rats. It was found that 6h after instillation with dust, neutrophils were present in greater numbers in the blood and bronchoalveolar spaces than in lung interstitium. After 24h, there appeared to be a larger number of neutrophils in the lung interstitium in dust-instilled animals than in saline-instilled controls. These results indicate that intratracheal instillation of grain dust initiates an acute inflammatory reaction, and that there is an initial influx of neutrophils into the air spaces of the lung followed by transit of these cells into the lung interstitium.     

The Effect of Caffeic Acid on Zearalenone-induced Ovarian Granulosa Cell Apoptosis in Mice

Ovarian granulosa cells provide a special microenvironment for follicle formation and maturation through interaction with oocytes and their own secretion. A variety of harmful stimuli can cause granulosa cell apoptosis and metabolic disorders, reduce the quality of oocytes and have a negative impact on embryo formation. Zearalenone (ZEA) is a common cause of ovarian granulosa cells injury in the livestock industry, which is produced by mycotoxins, and lack of effective treatment drug. Therefore, in the current study zearalenone was used to induce ovarian granulosa cell injury and to explore the protective effect of caffeic acid on zearalenone-induced ovarian granulosa cell apoptosis in mice. Mouse ovarian granulosa cells were isolated by mechanical method, and indirect immunofluorescence was used to identify the isolated cells. MTT assay was used to determine the effect of caffeic acid on the activity of normal mouse ovarian granulosa cells.After granulosa cells were co-treated with caffeic acid (200, 100 and 50 μg·mL^-1) and ZEA for 24 hours, and control and ZEA group were set up at the same time, cell morphology and adherence were observed under a microscope. MTT was also used to detect cell viability. Caspase-3 mRNA expression level was detected by qRT-PCR. Cleaved-caspase-3 and cleaved-PARP protein expre-ssion levels were determined by Western blot. The results showed that positive FSHR staining appeared in cell cytoplasm of the test group, which confirmed that the isolated cells were mouse ovarian granulosa cells. The cell viability was above 90% which showed that caffeic acid had no toxic effect on granulosa cells. Compared with control group, ZEA group had smaller cell size, poor adherence, increased cell gap, and significant reduction in cell viability (P<0.001). Furthermore, the relative expression of caspase-3 mRNA, and cleaved-caspase-3 and cleaved-PARP protein level were significantly increased (P<0.001) compared with the control group. After caffeic acid treatment, cell gap was reduced, adherence was tight, cell viability was significantly increased (P<0.001). Caffeic acid significantly reduced zearalenone-induced increase in caspase-3 mRNA, and cleaved-caspase-3 and cleaved-PARP protein expression level (P<0.001). This study indicated that caffeic acid can restore granulosa cell viability by inhibiting ZEA-induced apoptosis.     

Recognition and classification of dysmyelopoiesis in the dog: a review

Dysmyelopoiesis is defined as a hematologic disorder characterized by the presence of cytopenias in the blood and dysplastic cells in one or more hematologic cell lines in the blood or bone marrow. The causes of dysmyelopoiesis include acquired mutations in hematopoietic stem cells (i.e., myelodysplastic syndromes [MDSs]), congenital defects in hematopoiesis, and dysmyelopoietic conditions associated with various disease processes, drug treatments, or toxin exposure. Two major subtypes of MDSs (i.e., MDS with refractory cytopenias and MDS with excess myeloblasts) have been described that differ in clinical presentation, response to treatment, and survival time. The most frequently occurring causes of secondary dysmyelopoiesis include immune-mediated hematologic diseases, lymphoid malignancies, and exposure to chemotherapeutic drugs. Differentiation of the various causes of dysmyelopoiesis is essential for establishing an appropriate therapeutic plan and for determining prognosis.     

Canine Haematopoietic Chimerism Analyses by Semiquantitative Fluorescence Detection of Variable Number of Tandem Repeat Polymorphism

Canine models are successfully applied to the study of haematopoietic stem cell transplantation (HSCT). Monitoring of haematopoietic donor/recipient chimerism is of major significance in detecting and quantifying engraftment or graft rejection of the donor-derived haematopoietic cells after transplantation. Radioactive analyses of polymorphic microsatellite markers are commonly used for chimerism analyses. We describe an improved, non-isotopic method that is based on the analysis of microsatellite markers in donor and recipient cells using capillary electrophoresis and fluorescence detection. Artificial mixtures of donor and recipient DNA that were generated from peripheral blood mononuclear cells from dog leukocyte antigen-identical siblings were used to analyse the sensitivity of the assay. DNA from dogs that had received HSCT were also analysed in order to demonstrate the feasibility of the method in vivo. For chimerism analyses, six different microsatellite loci were systematically amplified using fluorescent PCR primer. The fluorescent polymerase chain reaction products were separated by capillary electrophoresis using POP4 on a 310 ABI Prism Genetic Analyzer. After electrophoresis, fluorescence signals were automatically sized and quantified using GeneScan software. The method described provides an accurate assessment of haematopoietic chimerism in the canine model with significantly reduced hands-on time compared to conventional gel electrophoresis.