Detecting interaction(s) and assessing the impact of component subsets in a chemical mixture using fixed-ratio mixture ray designs

An important environmental and regulatory issue is the protection of human health from potential adverse effects of cumulative exposure to multiple chemicals. Earlier literature suggested restricting inference to specific fixed-ratio rays of interest. Based on appropriate definitions of additivity, single chemical data are used to predict the relationship among the chemicals under the zero-interaction case. Parametric comparisons between the additivity model and the model fit along the fixed-ratio ray(s) are used to detect departure from additivity. Collection of data along reduced fixed-ratio rays, where subsets of chemicals of interest are removed from the mixture and the remaining compounds are at the same relative ratios as considered in the full ray, allow researchers to make inference about the effect of the removed chemicals. Methods for fitting simultaneous confidence bands about the difference between the best fitting model and the model predicted under additivity are developed to identify regions along the rays where significant interactions occur. This general approach is termed the “single chemicals required” (SCR) method of analysis. A second approach, termed “single chemicals not required” (SCNR) method of analysis, is based on underlying assumptions about the parameterization of the response surface. Under general assumptions, polynomial terms for models fit along fixed-ratio rays are associated with interaction terms. Consideration is given to the case where only data along the mixture rays are available. Tests of hypotheses, which consider interactions due to subsets of chemicals, are also developed.     

Statistical analysis of interactive cytotoxicity in human epidermal keratinocytes following exposure to a mixture of four metals

Exposure to mixtures of chemicals is an important and relevant environmental issue. Of particular interest is the detection and characterization of departure of biological effects from additivity. Methodology based on the assumption of additivity is used in fitting single-chemicaldata. Interactionsare determined and characterized by making comparisons between the observed and predicted responses at mixtures along a fixed ratio ray of the component substances. Two simultaneous tests are developed for testing for any departure from additivity. Multiple comparisons procedures are used to compare observed responses to that predicted under additivity. A simultaneous confidence band on the predicted responses along the mixture ray is also developed. The methods are illustrated with cytotoxicity data that arise when human epidermal keratinocytes are exposed to a mixture of arsenic, chromium, cadmium, and lead. Synergistic, antagonistic, and additive cytotoxicities were observed at different dose levels of the four-metal mixture.     

Subtotal pericardectomy and epicardial excision for treatment of coccidioidomycosis-induced effusive-constrictive pericarditis in dogs: 17 cases (1999-2003)

OBJECTIVE: To determine the history, clinicopathologic findings, and results of surgery for effusive-constrictive pericarditis associated with Coccidioides immitis infection in dogs. DESIGN: Retrospective study. ANIMALS: 17 client-owned dogs that underwent a subtotal pericardectomy and epicardial excision. PROCEDURE: Hospital records from May 1999 to June 2003 were reviewed. Data collected included history, clinicopathologic findings, treatments, and outcome. Follow-up information was obtained via recheck examination and by use of standardized telephone interviews with referring veterinarians and owners. RESULTS: All dogs were of large breeds, and most were male (mean age, 4.66 years). Ten dogs had no prior history of C. immitis infection, and 7 dogs had chronic infection with C. immitis. Having a chronic C. immitis infection reduced the odds of survival, compared with no previous infection. All dogs had clinical signs of right-sided heart failure. All dogs had serum titers (range, 1:8 to 1:256) for antibodies against C. immitis prior to surgery, and titers were not significantly associated with outcome. Predominant echocardiographic findings were thickened pericardium, reduced right ventricular filling, and pleural or pericardial effusion. All dogs underwent a subtotal pericardectomy and epicardial excision and had fibrosing pyogranulomatous pericarditis in biopsy specimens obtained during surgery. The perioperative mortality rate was 23.5%, and the 2-year postdischarge survival rate was 82%. CONCLUSIONS AND CLINICAL RELEVANCE: Surgical treatment via subtotal pericardectomy and epicardial excision is successful at relieving right-sided heart failure in dogs with effusive-constrictive pericarditis secondary to C. immitis infection, but long-term treatment with antifungal agents may still be required.     

Biochemical analysis of the articular cartilage and subchondral and trabecular bone of the metacarpophalangeal joint of horses with early osteoarthritis

OBJECTIVE: To assess whether site-related changes in biochemical composition are present in the cartilage and subchondral and trabecular bone of the metacarpophalangeal joint of horses with early osteoarthritis. SAMPLE POPULATION: Right metacarpophalangeal joints from 59 mature warmblood horses. PROCEDURE: Biochemical data (cross-link, amino acid, DNA, and ash contents; denatured collagen and glycosaminoglycan [GAG] concentrations; bone mineral density; and mineral composition) were obtained from 2 differently loaded sites of phalanx I cartilage and subchondral and trabecular bone samples; data were compared with previously published values from nonosteoarthritic equine joints. RESULTS: Compared with findings in nonosteoarthritic joints, GAG concentration was lower in cartilage from osteoarthritic joints and there was a loss of site differences in cellularity and lysylpyridinoline (LP) cross-link content. In subchondral bone, LP cross-link content was decreased overall and there was a loss of site differences in osteoarthritic joints; ash content was higher in the osteoarthritic joints. Hydroxyproline content in trabecular bone from osteoarthritic joints was greater than that in nonosteoarthritic trabecular bone. In all 3 layers and at both sites, the linear increase of the pentosidine cross-link content with age had diminished or was not apparent in the horses with osteoarthritic joints. CONCLUSIONS AND CLINICAL RELEVANCE: In equine metacarpophalangeal joints with early osteoarthritis, distinct biochemical changes were detected in the cartilage and subchondral and trabecular bone. The dissimilarity in response of the different tissues and differences between the sites that are affected may be related to differences in biomechanical loading and transmission and dissipation of force.     

Comparison of three methods for arthrodesis of the distal intertarsal and tarsometatarsal joints in horses

OBJECTIVE: To evaluate the effects of diode laser surgery (LS), surgical drilling (SD), and intraarticular sodium monoiodoacetate (MIA) as methods for fusing the distal intertarsal (DIT) and tarsometatarsal (TMT) joints in horses. STUDY DESIGN: Experimental study. ANIMALS: Adult horses (15) without radiographic signs of osteoarthritis (OA) of the DIT and TMT joints. METHODS: Group 1 (n=3) had LS performed bilaterally on DIT and TMT joints; 1 horse was evaluated for 1 week and 2 horses were evaluated for 2 weeks. Group 2 (n=6) had LS on DIT and TMT joints of 1 tarsus and MIA administration into the contralateral DIT and TMT joints and were evaluated for 6 months. Group 3 (n=6) had LS performed on DIT and TMT joints of 1 tarsus and SD of the contralateral DIT and TMT joints and were evaluated for 12 months. Postoperative comfort, lameness, radiography, microradiography, and histology scores were compared using repeated measures ANOVA, and paired or 2 sample t-tests; significance was set at P<.05. RESULTS: LS caused the least postoperative morbidity. In group 2, horses were less lame in 4 LS-treated limbs and 2 MIA-treated limbs at 6 months when compared with the contralateral limb. In group 3, horses were less lame in 5 LS-treated limbs and 1 SD-treated limb at 6 and 12 months compared with the contralateral limb. On microradiography, 11 MIA joints and 2 LS joints had bone bridging the joint at 6 months whereas 8 SD joints and 5 LS joints had bone bridging at 12 months. Significantly more joint space was bridged by bone in MIA- (51.4%) and SD (46.2%)-treated joints compared with LS joints at 6 (30.6%) and 12 (28.5%) months, respectively (P<.05). CLINICAL RELEVANCE: SD and MIA resulted in more bone bridging of the distal 2 tarsal joints, than LS. However, LS seemingly caused less pain and discomfort to horses in the immediate postoperative period; horses were generally less lame in the LS limb. More laser energy may need to be applied to these joints to promote fusion; however, it may also have beneficial effects beyond fusion. Further research on horses with OA of the distal 2 tarsal joints is needed to determine whether LS can cause soundness without facilitating bony fusion.     

Accuracy and optimization of force platform gait analysis in Labradors with cranial cruciate disease evaluated at a walking gait

OBJECTIVE: To determine the combination of ground reaction forces (GRFs) that best discriminates between lame and non-lame dogs. To compare the sensitivity of force platform gait analysis and visual observation at detecting gait abnormalities in Labradors after surgery for rupture of the cranial cruciate ligament (CCL). ANIMALS: All dogs were adult Labrador Retrievers: 17 free of orthopedic and neurologic abnormalities, 100 with unilateral CCL rupture, and 131 studied 6 months after surgery for unilateral CCL injury, 15 with observable lameness. PROCEDURE: Dogs were walked over a force platform with GRF recorded during the stance phase. Analytic properties of force platform gait analysis were calculated for several combinations of forces. The probability of visual observation detecting a gait abnormality was compared with that of force platform gait analysis. RESULTS: We determined that a combination of peak vertical force (PVF) and falling slope were optimal for discriminating sound and lame Labradors. After surgery, many dogs (75%) with no observable lameness failed to achieve GRFs consistent with sound Labradors. CONCLUSION: A force platform is an accurate method of assessing lameness in Labradors with CCL rupture and is more sensitive than visual observation. Assessing lameness with a combination of GRFs is better than using univariate GRFs. CLINICAL RELEVANCE: Therapies for stifle lameness can be accurately and objectively evaluated using 2 vertical ground reaction forces obtained from a force platform.     

Canine sacroiliac luxation: anatomic study of dorsoventral articular surface angulation and safe corridor for placement of screws used for lag fixation

OBJECTIVE: To define a safe corridor in the dorsoventral plane to facilitate placement of screws inserted in lag fashion within the sacral body for fixation of sacroiliac fracture-luxation injuries in dogs. STUDY DESIGN: Anatomic study. SAMPLE POPULATION: Cadaveric canine sacra. METHODS: Canine sacra (n=45) were used for a radiographic study to define a safe corridor in the dorsoventral plane for placement of screws inserted in lag fashion for fixation of sacroiliac luxation in the dog. The defined safe corridor allowed drilling to a depth of 65% of the sacral width to ensure screw purchase of > or =60%. Effects of positioning and measurement techniques were evaluated. RESULTS: Eighty-seven safe corridors were measured. The mean articular surface was 100+/-4.52 degrees from horizontal. Mean maximum, optimum, and minimum safe corridor drill angles were 111+/- 4.57 degrees, 100+/-4.70 degrees, and 89+/-5.17 degrees, respectively, from the articular surface. Predicted surgeon error of +/-4 degrees was used to define the safe corridor for use clinically. CONCLUSIONS: In 91% of sacra, a drill angle of 100+/-4 degrees would remain ventral to the vertebral canal. Twelve sacra (14%) were at risk of penetration of the pelvic canal. A drill angle of 97+/-4 degrees avoids penetration of the vertebral canal in all sacra measured but risks ventral exit from the body in 30% of sacra studied. CLINICAL RELEVANCE: A drill angle of 97 degrees from the articular surface is recommended for insertion of screws for lag fixation of canine sacroiliac luxation.     

The epidemiology and diagnosis of bluetongue with particular reference to Corsica

Bluetongue (BT) and/or BT viruses (BTV) have been identified in the Mediterranean basin and the Balkans each year from 1998 to 2002 and in particular BTV serotype 2 in the French Island of Corsica (2000 and 2001). In response to these virus incursions, the French Veterinary Authorities carried out epidemiological studies that included virological, serological and entomological analysis, and two vaccination campaigns performed in the winter of 2000/2001 and the winter and spring of 2001 and 2002. Rapid and reliable serotype differentiation is essential at the start of an outbreak to allow an early selection of vaccine to control the spread of the virus. Thus, molecular tools, that complement conventional methods, have been developed for early detection of infection, determination of the serotype, and differentiation between natural infection and vaccination. Serological results showed that the first vaccination campaign during the winter of 2000/2001 did not provide full protection for all sheep and during the summer of 2001, 335 sheep flocks in Corsica were again infected by BTV 2 (7-fold more that in 2000). Entomological studies have demonstrated that the only proven vector of the disease, Culicoides imicola, was present in the island in 2000 and that it has successfully established itself in Corsica. The safety and immunogenicity of the commercial South African vaccine were studied. Fourteen sheep were vaccinated and then observed for clinical signs. Blood, sera, spleen and lymph nodes were collected and analyzed, and the results confirmed the safety and potency of using this vaccine to protect sheep from clinical disease. As a result, an intensive vaccination campaign was performed during winter and spring 2001/2002. No cases of BT had been observed by the end of summer 2002, indicating that the vaccination campaign has been successful in protecting sheep from infection.     

Delivery of growth factors using gene therapy to enhance bone healing

OBJECTIVE: To review the delivery of growth factors using gene therapy for enhancing long-bone fracture healing. STUDY DESIGN: Literature review. METHODS: MEDLINE and CAB Abstracts literature search (1980-2004). RESULTS: Non-union and infected non-union are relatively common complications of long-bone fractures in human and veterinary patients. Growth factors are cytokines that regulate many cell functions important in fracture healing. Exogenous growth factors can be delivered to the fracture site as recombinant proteins or using gene therapy. Recombinant human bone morphogenetic protein-2 and -7 (rhBMP-2 and -7), in particular, enhance fracture healing in numerous experimental and clinical studies. Some limitations with use of recombinant proteins may be overcome by use of gene therapy. Gene therapy involves delivery of the growth factor gene to cells at the fracture site using a viral or non-viral vector. The gene is then expressed (protein synthesis) by cells at the fracture site. Delivery of the BMP gene to the fracture site using gene therapy has been evaluated in laboratory animal models of non-union, with favorable results and without complications. CONCLUSION: Delivery of growth factors, particularly members of BMP family, to the fracture site using gene therapy may be a method to enhance fracture healing. Use of this technology may improve the prognosis for patients with long-bone fractures. CLINICAL RELEVANCE: Clinical application of gene therapy could improve the prognosis for human and veterinary patients with long-bone fractures, but has not been evaluated clinically.     

Exhaled breath condensate (EBC) collection in cats--description of a non-invasive technique to investigate airway disease

Exhaled breath condensate has been collected in other species and used as a non-invasive method of evaluating airway disease by measurement of various markers in the fluid, including hydrogen peroxide, nitric oxide, leukotrienes and prostaglandins. We describe a novel technique for the collection of exhaled breath condensate from cats, which enabled collection of fluid and measurement of its hydrogen peroxide concentration. Further studies will be needed to establish the value of this technique in the investigation of feline respiratory disease.