EVALUATION OF PLASMA TIME-ACTIVITY CURVES OF TECHNETIUM-99M-MEBROFENIN FOR MEASUREMENT OF HEPATIC FUNCTION IN DOGS

In this study, plasma time-activity curves of 99mTc-mebrofenin were used to quantify hepatic function in dogs before and after induction of hepatic damage using a hepatotoxic agent. Nine dogs were determined to be healthy on the basis of physical examination, laboratory data and hepatic imaging. Plasma samples were collected 1, 3, 5, 7, 9, 15, 20, 30, 40, 50, and 60 minutes following a peripheral venous injection of 111-222 MBq (3-6 mCi) of 99mTc-mebrofenin. The area under the plasma time-activity curve (AUC) was calculated using two different methods and compared to direct measurement of the hepatic extraction efficiency. First pass hepatic extraction efficiency of 99mTc-mebrofenin was calculated from differential equation analysis of a two-compartment model following mesenteric venous injection of the radiopharmaceutical. In 7 of the original 9 dogs and 2 additional healthy dogs, plasma clearance and hepatic extraction efficiency determination were repeated following induction of hepatic injury by thiacetarsamide (3 mg/kg IV twice daily for 1 day). In one additional dog, hepatic injury was induced using carbon tetrachloride (0.3 ml/kg IP). Plasma time-activity curves of 99mTc-mebrofenin had kinetics of a two compartment model. Area under the curve was highly correlated with hepatic extraction efficiency. The AUC integrated from 1-60 minutes (AUC60) had the best correlation with hepatic extraction efficiency (r2 = 0.978, p < 0.001). A formula for calculation of hepatic extraction efficiency was derived using linear regression analysis: hepatic extraction efficiency = 105.583 - 3.099 x 10(5) x AUC60. Plasma clearance of a peripheral venous injection of 99mTc-mebrofenin is a simple, non-invasive, convenient method to quantify hepatic function which can be performed without a gamma camera.     

Single layer centrifugation of fresh dromedary camel semen improves sperm quality and in vitro fertilization capacity compared with simple sperm washing

Single layer centrifugation (SLC) through a colloid is a tool for selecting viable mammalian spermatozoa but has not been used previously for fresh dromedary camel sperm. Semen from six camels (2 ejaculates/male) was diluted 1:5 (v:v) or 1:10 (v:v) in a Tris–citrate–fructose buffer for mechanical liquefaction by gentle pipetting. Following liquefaction, semen was processed either by SLC or by centrifugation without a colloid (control). Total and progressive motilities, CASA kinematics, vitality and acrosome integrity (eosin–nigrosin) and plasma membrane integrity (Hypo‐osmotic swelling test; HOST), and fertilizing ability in a heterologous assay (zona‐free goat oocytes) were evaluated. Both total (p = .003) and progressive motilities (p = .003) were higher in SLC‐processed than in control semen samples, irrespective of dilution. Positive HOST values increased when using colloid in 1:5 (p = .001) and 1:10 dilution (p = .010). Colloid‐selected sperm had higher penetration rates than controls (p < .001 and p = .02 for 1:5 and 1:10 dilutions, respectively). However, only the SLC sperm at 1:5 dilution showed higher percentages of pronuclear formation (p = .02) than controls. Dilution effect was only significant for total motility before in vitro fertilization, with higher values for the 1:5 dilution (p = .033). The recovery rates of motile sperm between dilutions were similar (26.1% vs 35.4%; p = .226). In conclusion, SLC is a promising tool for selecting functional dromedary camel sperm and warrants more research.     

Cytology of Infection of Maize Seedlings by Fusarium moniliforme and Immunolocalization of the Pathogenesis-Related PRms Protein

ABSTRACT We have investigated the histology of infection of maize seedlings by Fusarium moniliforme in association with a biochemical host defense response, the accumulation of the PRms (pathogenesis-related maize seed) protein. Light microscopy of trypan blue-stained sections and scanning electron microscopy revealed direct penetration by F. moniliforme hyphae through the epidermal cells of the seedling and colonization of the host tissue by inter- and intracellular modes of growth. Pathogen ingress into the infected tissue was associated with the induction of defense-related ultrastructural modifications, as exemplified by the formation of appositions on the outer host cell wall surface, the occlusion of intercellular spaces, and the formation of papillae. Cellular and subcellular immunolocalization studies revealed that PRms accumulated at very high levels in those cells types that represent the first barrier for fungal penetration such as the aleurone layer of germinating seeds and the scutellar epithelial cells of isolated germinating embryos. A highly localized accumulation of PRms within papillae of the inner scutellar parenchyma cells also occurred, suggesting that signaling mechanisms that lead to the accumulation of PRms in papillae of cell types that are distant from the invading pathogen must operate in the infected maize tissues. Our study also revealed the presence of a large number of fungal cells with an abnormal shape that showed PRms-specific labeling. PRms was found to accumulate in clusters over the fungal cell wall. Taken together, the occurrence of PRms in cell types that first establish contact with the pathogen, as well as in papillae, and in association with fungal cell walls suggests that PRms may have a function in the plant defense response.     

Lymphocyte Populations in Joint Tissues from Dogs with Inflammatory Stifle Arthritis and Associated Degenerative Cranial Cruciate Ligament Rupture

Objective: To evaluate lymphocyte populations in stifle synovium and synovial fluid of dogs with degenerative cranial cruciate ligament rupture (CCLR). Study Design: Prospective clinical study. Animals: Dogs (n=25) with stifle arthritis and CCLR, 7 dogs with arthritis associated with cartilage degeneration (osteoarthritis [OA]), and 12 healthy Beagle dogs with intact CCL. Methods: Arthritis was graded radiographically in CCLR dogs. After collection of joint tissues, mononuclear cells were isolated and subsequently analyzed using flow cytometry for expression of CD3, CD4, CD8, and CD21. Results: The proportions of CD4⁺ T helper lymphocytes, CD8⁺ cytotoxic T lymphocytes, and CD3⁺CD4^?CD8^? T lymphocytes were increased in synovium from dogs with CCLR compared with synovium from healthy Beagle dogs (P<.05). The proportion of CD3⁺CD4^?CD8^? T lymphocytes in synovial fluid was increased in dogs with CCLR compared with dogs with OA (P<.05). In dogs with CCLR, the proportion of CD3⁺CD4^?CD8^? T lymphocytes in synovial fluid was inversely correlated with radiographic arthritis (S_R=?0.68, P<.005). Conclusion: Lymphocytic inflammation of stifle synovium and synovial fluid is an important feature of the CCLR arthropathy. Lymphocyte populations include T lymphocytes expressing CD4 and CD8, and CD3⁺CD4^?CD8^? T lymphocytes. Presence of CD3⁺CD4^?CD8^? T lymphocytes was associated with development of stifle synovitis. Further work is needed to fully identify the phenotype of these cells.     

Evaluation of Indicators of Weight-Carrying Ability of Light Riding Horses

To answer the question of whether horse height, cannon bone circumference, and loin width can be used as indicators of weight-carrying ability in light horses, eight mature horses underwent a submaximal mounted standard exercise test under four conditions: carrying 15, 20, 25, or 30% of their body weight. Heart rate was monitored, plasma lactate concentration was determined in jugular blood samples pre-exercise, immediately post-exercise, and 10 minutes post-exercise, with serum creatine kinase activity determined at the same times as plasma lactate concentrations, with additional samples collected at 24 hours and 48 hours post-exercise. Muscle soreness and muscle tightness scores were determined using a subjective scoring system 24 hours before and 24 hours after exercise. Heart rates remained significantly higher when the horses carried 25 and 30% of their body weight. Plasma lactate concentrations immediately and 10 minutes after exercise differed when horses carried 30% of their body weight compared with 15, 20, and 25% weight carriage. Horses tended to have a greater change in muscle soreness and muscle tightness when carrying 25% of their body weight, and a significant change in soreness and tightness scores was found in horses carrying 30% of their body weight. Loin width and cannon bone circumference were found to be negatively correlated to the changes in muscle soreness and tightness scores. In conclusion, the data suggest that horses with wider loin and thicker cannon bone circumference became less sore when carrying heavier weight loads.     

QUANTITATIVE THYROID SCINTIGRAPHY AS A PREDICTOR OF SERUM THYROXIN CONCENTRATION IN NORMAL AND HYPERTHYROID CATS

Quantitative thyroid scintigraphy using pertechnetate was performed in 43 cats with various T4 concentrations and compared to eight normal control cats. Quantitative parameters included percentage dose uptake of the radioisotope by the thyroid, thyroid:salivary ratio and rate of thyroid uptake. All cats were anesthetized for the scan, and images were obtained using both low-energy all purpose (LEAP) and pinhole collimators. All quantitative parameters were significantly correlated to the serum T4 concentration, but the best correlation was obtained using the 20-minute thyroid:salivary ratio using only the most intense of the two thyroid lobes. The thyroid:salivary ratio was a good predictor of the metabolic status of the thyroid.