Elizabeth M. Santschi

Normalized cDNA libraries from a porcine model of orthopedic implant-associated infection

Abstract. Staphylococcal infections that result from an alteration in a patients immune response at the surgical site are a major problem in procedures that incorporate biomaterials in trauma surgery and joint replacement. Diagnosis of infection based on pathogen detection is difficult and exacerbated by increasing numbers of partially or totally resistant strains of nosocomial pathogens, particularly Staphylococcus aureus. Expression profiling of a hosts cellular immune response could facilitate the identification of the pathways involved in pathogen recognition and eradication and could lead to more rational design of drugs and therapies. To this end, we constructed and characterized ten individually tagged and directionally cloned cDNA libraries from peripheral blood cells (PBC), spleen (Sp), thymus (Th), lymph node (LN), and bone marrow (BM) from immunologically naive and challenged pigs as part of an implant-associated orthopedic model of deep infection. Three of these libraries were normalized at C0t values 5, 10, 20, and 30. The libraries comprise more than 20 million primary transformants with an average insert length >1.4 kb. Cluster analysis of 7620 ESTs revealed 1029 clusters containing an average of 3.6 sequences and 3846 singletons. Gene discovery is estimated to be ∼64%. Searches of public databases resulted in 49.3% annotated porcine sequences, of which 22.2% had significant homologies to ESTs from a variety of species, and 28.5% were without a significant match in any public database. We also identified 9.1% ESTs as involved in host cell and organism defense and 11.5% related to cell signaling and communication. These sequences, together with the 28.5% appearing as novel, are of specific interest to the infectious disease process.

Influence of exercise on the distribution of technetium Tc 99m medronate following intra-articular injection in horses

OBJECTIVE To determine the effects of exercise on the distribution and pharmacokinetics of technetium Tc 99m medronate ((99m)Tc-MDP) following intra-articular (IA) injection in horses. ANIMALS 5 horses. PROCEDURES 1 antebrachiocarpal joint (ACJ)/horse was assigned to the exercised group (n = 5), and the contralateral ACJ was evaluated in the nonexercised group (5) after a minimum washout period of 7 days. Following IA injection of (99m)Tc-MDP (148 MBq), blood and scintigraphic images of the carpus were obtained at 5, 10, 15, 20, 25, 30, 45, 60, 90, 120, 240, 360, 480, 600, 720, and 1,440 minutes. Plasma and scintigraphic radioactivity were determined over time, and pharmacokinetic parameters were generated via noncompartmental and compartmental analyses. Each horse was monitored via physical and lameness examination and ACJ synovial fluid analysis before injection and at days 1, 2, 3, and 7. RESULTS Lameness was not observed. Mean ± SD synovial fluid WBC count increased at day 1 (exercised, 721 ± 234 cells/μL; nonexercised, 948 ± 223 cells/μL), but returned to baseline at days 3 and 7 Mean time to maximum plasma radioactivity was earlier in the exercised group (16.00 ± 2.35 minutes) than the nonexercised group (43.75 ± 3.64 minutes). Linear regression of the scintigraphic radioactivity-time curves revealed a greater negative slope in the exercised group within the first 25 minutes. There was no difference in absorption or elimination rate constants in a 2-compartment model. CONCLUSIONS AND CLINICAL RELEVANCE IA injection of (99m)Tc-MDP was safe and effective for evaluating synovial solute distribution. Exercise significantly increased early transfer of (99m)Tc-MDP from the ACJ into plasma, although absorption and elimination rate constants were not affected. Exercise may affect synovial clearance and withdrawal times of medications administered IA.

Surgical findings and outcome for dairy cattle with jejunal hemorrhage syndrome: 31 cases (2000-2007).

OBJECTIVE To describe signalment; surgical findings; short-, medium-, and long-term outcome; and recurrence rate for cattle undergoing celiotomy because of jejunal hemorrhage syndrome (JHS) and to analyze risk factors associated with outcome and recurrence. DESIGN Retrospective case series. ANIMALS 31 dairy cattle with JHS. PROCEDURES Medical records were analyzed. Follow-up information was obtained from owners of cattle surviving until discharge. RESULTS 18 of 31 (58%) cattle undergoing celiotomy survived to initial discharge. Fifteen (48%) and 13 (42%) were alive 6 and 12 months after discharge, respectively. All 5 deaths within 12 months after discharge were attributed to JHS recurrence. Survival time was 12 to 85 months for the 13 long-term survivors. Six of 7 that died > 12 months after celiotomy did so for reasons unrelated to JHS. Recurrence rate among short-term survivors was 7 of 18; 1 of these survived long-term. A significant proportion of affected cattle were Brown Swiss, compared with proportions for other breeds. Manual massage of the bowel to break down clots was associated with a significantly higher short-term survival rate than was enterectomy or enterotomy. Medium- and long-term survival rate was higher in cattle referred 24 to 48 hours after onset of signs. Length of obstructing blood clots was not associated with outcome. Other factors were not significantly associated with recurrence. CONCLUSIONS AND CLINICAL RELEVANCE Survival rates were higher than those in other reports. Prompt celiotomy and resolution by use of manual massage were associated with higher survival rates. In this population, JHS recurred in 7 of 18 short-term survivors.

Identification and radiation hybrid mapping of members of the porcine proteasome/ubiquitin system

We report the identification and radiation hybrid mapping of members of the proteasome/ubiquitin system in pigs that, so far, have only been identified in humans and cattle. Expressed sequence tags (ESTs) were constructed from ten oligo(dT)-primed individually tagged, directionally cloned and normalized cDNA libraries from peripheral blood cells (PBC), spleen (Sp), thymus (Th), lymph node (LN) and bone marrow (BM) from immunologically naive and challenged pigs as part of an implant-associated orthopedic infection model. The ESTs mapped using the 7000 rad IMpRH panel (Hawken et al., 1999) were ubiquitin fusion-degradation 1 like protein (UFD1L), ubiquitin activating enzyme E1 and ubiquitin-S27a fusion protein which mapped to porcine chromosomes 14, 7 and X, respectively.