Designed protein-protein association

The analysis of natural contact interfaces between protein subunits and between proteins has disclosed some general rules governing their association. We have applied these rules to produce a number of novel assemblies, demonstrating that a given protein can be engineered to form contacts at various points of its surface. Symmetry plays an important role because it defines the multiplicity of a designed contact and therefore the number of required mutations. Some of the proteins needed only a single side-chain alteration in order to associate to a higher-order complex. The mobility of the buried side chains has to be taken into account. Four assemblies have been structurally elucidated. Comparisons between the designed contacts and the results will provide useful guidelines for the development of future architectures.     

Application of a Balanced Scorecard System for Supporting Decision-Making in Contaminated Sites Remediation

This work introduces the development and application of a multi-goal evaluation procedure for assessing sets of remedial options that have previously been found technically suitable for managing a certain contaminated site. The construction of this decision support tool, in the form of a balanced scorecard, has followed a multi-attribute conception of the contaminated sites problem, identifying the most critical aspects considered by all stakeholders involved in contaminated sites management decisions. The developed tool has been checked in its formulation and application against an export benchmark of an Austrian remediation project. The performed evaluation allowed to identify and suggest a remedial option that best fitted the expectations of all stakeholders involved in the project. Furthermore, through the accomplishment of a sensitivity analysis it has been demonstrated that the system is stable and robust in terms of the chosen elicitation schemes, and that the preferences of the different stakeholders are influencing factors on the final result. The results and conclusions of this work present the benefits of implementing the developed balanced scorecard as a tool for supporting decision making in contaminated sites remediation projects.     

Effects of salinity, aeration and light intensity on oil globule absorption, feeding incidence, growth and survival of early-stage grouper Epinephelus coioides larvae

ABSTRACT: A series of experiments were conducted to examine the effects of salinity, aeration and light intensity on oil globule absorption, feeding incidence, and growth and survival of early-stage Epinephelus coioides larvae. Newly hatched larvae were transferred to 40-L aquaria at a density of 1500 individuals/aquarium. Larvae were exposed to different levels of aeration (0 mL/min per L, 0.62 mL/min per L, 1.25 mL/min per L, 2.50 mL/min per L, or 3.75 mL/min per L); salinity (8 ppt, 16 ppt, 24 ppt, 32 ppt, or 40 ppt); and light intensity (0 lx, 120 lx, 230 lx, 500 lx, or 700 lx) for 4–6 days. Twenty larvae were sampled daily at 11:00 hours to measure for total length (TL), oil globule volume, and feeding incidence. Survival rates were determined by counting the total number of larvae remaining in each aquarium at the end of the experiment. Significantly higher survival rates ( P   <  0.05) were observed at aeration levels of 0.62 mL/min per L and 1.25 mL/min per L, at salinity levels of 16 ppt and 24 ppt, and at light intensities of 500 lx and 700 lx. The influence of aeration level, salinity and light intensity on oil globule absorption, feeding incidence, and growth and survival of early-stage grouper larvae are discussed.     

MAGNETIC RESONANCE IMAGING OF THE BRAIN OF NORMAL NEONATAL FOALS

Magnetic resonance imaging (MRI) was performed on gthe brain of 5 normal, anesthetized, neonatal (age 3-to-6 days) Quarter Horse foals. The objectives of the study were to develop a technique for imaging the brain of neonatal foals, and to ascertain their normal brain anatomy. Interavenous propofol was administered for induction and maintenance of general anesthesia. Using spin echo MR techniques, T1 weighted sagittal and transverse views, and spin density and T2 weighted transverse views were successfully made of each foal. MR images provided excellent visualization of many anatomic struictures of the brain and head. MRI of the bgrain is feasible for selected neonantal equine patients.     

Repetitive propofol administration in dogs and cats

A bolus of propofol was administered to 10 dogs (6 mg/kg intravenously [IV]) and 10 cats (10 mg/kg IV) on three consecutive days. The occurrence of apnea, heart and respiratory rates, blood pressure, time to movement, and changes in a complete blood count and biochemical profile were recorded. Apnea was not seen in the dogs but was seen in three cats. Slight increases in the number of Heinz bodies were seen in six cats, but the increases were not considered clinically significant. No apparent cumulative adverse effects were seen from a bolus of bisulfite-containing propofol, administered on three consecutive days.     

Role of lymph-borne cells in the early stages of scrapie agent dissemination from the skin

Scrapie is a natural transmissible spongiform encephalopathy (TSE) of sheep, infecting the animal via the gastrointestinal tract or the skin. This project tested the hypotheses that lymph-borne cells (especially dendritic cells) are crucial for the systemic dissemination of the infectious agent from the site of infection in the skin, that PrP genotype affects PrPSC association with dendritic cells and that PrPSC carriage by cells affects their expression of cytokines. Skin, of scrapie-susceptible VRQ/ARR and scrapie-resistant ARR/ARR PrP genotypes, was scarified with FITC-labelled PrPSC. Pseudoafferent lymphatic cannulation was then used to monitor the presence of FITC-PrPSC over time in different lymph cell populations and plasma in the draining afferent lymphatics. The major observation was that PrPSC did not associate significantly with any lymphocyte or dendritic cell population in the 5 days following PrPSC scarification. The only cells seen to associate with PrPSC were neutrophils. Furthermore, despite the quantity of PrPSC used for scarification being equivalent to a standard infectious dose (the VRQ/ARR sheep dying at approximately 260 days) the only PrP found in afferent lymph during the 0-5-day period was proteinase K sensitive (i.e. soluble PrPC). No differences were observed between the PrP genotypes. Analysis of the effects of PrPSC scarification of cellular cytokine mRNA expression (by a nuclease protection assay) showed raised levels of IL-1beta and IL-8 in the susceptible VRQ/ARR group and raised levels of IFNgamma in the resistant ARR/ARR animals.     

Pharmacokinetics and clinical effects of a subanesthetic continuous rate infusion of ketamine in awake horses

OBJECTIVE: To determine the pharmacokinetics and clinical effects of a subanesthetic, continuous rate infusion of ketamine administered to healthy awake horses. ANIMALS: 8 adult horses. PROCEDURES: Ketamine hydrochloride was administered to 2 horses, in a pilot study, at rates ranging from 0.4 to 1.6 mg/kg/h for 6 hours to determine an appropriate dose that did not cause adverse effects. Ketamine was then administered to 6 horses for a total of 12 hours (3 horses at 0.4 mg/kg/h for 6 hours followed by 0.8 mg/kg/h for 6 hours and 3 horses at 0.8 mg/kg/h for 6 hours followed by 0.4 mg/kg/h for 6 hours). Concentration of ketamine in plasma, heart rate, respiratory rate, blood pressure, physical activity, and analgesia were measured prior to, during, and following infusion. Analgesic testing was performed with a modified hoof tester applied at a measured force to the withers and radius. RESULTS: No signs of excitement and no significant changes in the measured physiologic variables during infusion rates of 0.4 and 0.8 mg of ketamine/kg/h were found. At 6 hours following infusions, heart rate and mean arterial pressure were decreased, compared with preinfusion measurements. An analgesic effect could not be demonstrated during or after infusion. Pharmacokinetic variables for 0.4 and 0.8 mg/kg/h infusions were not significantly different. CONCLUSIONS AND CLINICAL RELEVANCE: Ketamine can be administered to awake horses at 0.4 or 0.8 mg/kg/h without adverse behavioral effects. The observed pharmacokinetic values are different than those reported for single-dose IV bolus administration of this drug.     

Musculoskeletal Corynebacterium pseudotuberculosis infection in horses: 35 cases (1999-2009)

Objective-To describe the clinical course and outcome in horses in which Corynebacterium pseudotuberculosis infections were associated with musculoskeletal disease and lameness. Design-Retrospective case series. Animals-35 horses. Procedures-Clinical and clinicopathologic data were collected from horses diagnosed with lameness associated with C pseudotuberculosis infection between 1999 and 2009. Results-32 (91.4%) horses had grade 4/5 lameness. Three (8.6%) horses had grade 5/5 lameness. Abscesses were diagnosed by clinical or ultrasonographic examination. Abscesses were located in the axillary or triceps region in 25 (71.4%) horses, the stifle region in 2 (5.7%), and the popliteal lymph node in 1 (2.9%). Diffuse lymphangitis was seen in 4 (11.4%) horses, osteomyelitis in 2 (5.7%) horses, and septic arthritis in 2 (5.7%) horses. Horses commonly had clinicopathologic abnormalities characterized by neutrophilia (96.4%), anemia (67.8%), hypoalbuminemia (66.6%), or hyperfibrinogenemia (42.8%). Treatment included surgical drainage of the abscess in 21 (60%) horses, performed under ultrasonography in 20 horses; anti-inflammatory medications in 34 (97.1 %) horses; and antimicrobials in 30 (85.7%) horses. Conclusions and Clinical Relevance-C pseudotuberculosis infection of the limbs in horses typically results in severe lameness but may have a favorable prognosis. The diagnosis may be challenging, and results of blood work consistent with inflammation are nonspecific, but anemia, hyperglobulinemia, and increased synergistic hemolysis inhibition titers are common. Ultrasonography may localize the lesions and facilitate surgical drainage to alleviate lameness. When C pseudotuberculosis musculoskeletal infection results in osteomyelitis or septic arthritis, the prognosis for survival is poor.