Use of an antineoepitope antibody for identification of type-II collagen degradation in equine articular cartilage.

OBJECTIVE: To develop an antibody that specifically recognizes collagenase-cleaved type-II collagen in equine articular cartilage. SAMPLE POPULATION: Cartilage specimens from horses euthanatized for problems unrelated to the musculoskeletal system. PROCEDURE: A peptide was synthesized representing the carboxy- (C-) terminus (neoepitope) of the equine type-II collagen fragment created by mammalian collagenases. This peptide was used to produce a polyclonal antibody, characterized by western analysis for reactivity to native and collagenase-cleaved equine collagens. The antibody was evaluated as an antineoepitope antibody by ELISA, using peptides +/- an amino acid at the C-terminus of the immunizing peptide. Collagen cleavage was assayed from equine articular cartilage cultured with interleukin-1 (IL-1), +/- a synthetic MMP inhibitor, BAY 12-9566. Cartilage specimens from osteoarthritic and nonarthritic joints were compared for antibody staining. RESULTS: An antibody, 234CEQ, recognized only collagenase-generated 3/4-length fragments of equine type-II collagen. This was a true antineoepitope antibody, as altering the C-terminus of the immunizing peptide significantly decreased competition for binding in an inhibition ELISA. The IL-1-induced release of type-II collagen fragments from articular cartilage was prevented with the MMP inhibitor. Cartilage from an osteoarthritic joint of a horse had increased staining with the 234CEQ antibody, compared with normal articular cartilage. CONCLUSIONS AND CLINICAL RELEVANCE: We generated an antineoepitope antibody recognizing collagenase-cleaved type-II collagen of horses. This antibody detects increases in type-II collagen cleavage in diseased equine articular cartilage. The 234CEQ antibody has the potential to aid in the early diagnosis of arthritis and to monitor treatment responses.     

Use of laser photocoagulation for treatment of cutaneous angiomatosis in one dog and two cats.

One dog and 2 cats were evaluated because of multiple progressively enlarging cutaneous vascular plaques. Biopsies were performed and revealed small well-circumscribed dermal nodules of dilated fully enclosed blood-filled spaces lined by single layers of endothelial cells aligned on thin fibrous walls, with minimal mitotic activity. A diagnosis of cutaneous angiomatosis was made in all 3 animals. Cutaneous angiomatosis is a progressive proliferative lesion of vascular tissue involving the dermis and subcutaneous tissues of dogs and cats. Lasers of selected wavelengths have been used to induce photothermal coagulation of hemoglobin-containing cutaneous lesions in humans; argon-pumped dye and neodymium:yttrium-aluminum-garnet lasers were used to treat the cutaneous lesions in the animals of this report. Laser treatment may provide an alternative to wide surgical resection or limb amputation for management of this condition in veterinary patients.     

Immunohistochemical Detection of Mycoplasma agalactiae in Formalin‐Fixed,Paraffin‐Embedded Tissues from Naturally and Experimentally Infected Goats

Samples from the mammary tissue of 14 lactating goats (12 naturally infected and two experimentally infected) were examined for the presence of Mycoplasma agalactiae. A monoclonal antibody (5G12) was applied to formalin‐fixed, paraffin‐wax‐embedded sections and labelled by the avidin–biotin peroxidase complex (ABC) method. Histological examination of tissue sections revealed strong immunoreactivity in all animals included in the study. Mycoplasma agalactiae antigen was mainly detected in the cellular debris at the periphery of purulent exudates present within lactiferous sinuses, and lactiferous and interlobular ducts. In addition, M. agalactiae organisms appeared in the cytoplasm of the epithelium of ducts, and in infiltrating macrophages and neutrophils within the ducts, alveoli, interstitial tissue and regional lymph node sinuses. It is concluded that this monoclonal antibody‐based immunohistochemical technique is an efficient and specific method for the post‐mortem detection of M. agalactiae in cases of clinical mastitis as well as being a useful tool for the study of the route of infection and cellular types involved during mastitis caused by this organism.     

Application and crop safety parameters for soil fumigants

Metam sodium alone and in combination with 1,3-dichloropropene plus 17% chloropicrin (1,3-D+C-17) were evaluated under polyethylene mulch film as alternatives for methyl bromide in tobacco and tomato transplant production for both efficacy against pests and crop safety. Eight different weed species, 10 genera or species of fungi and several agronomic criteria were evaluated at three different sites. In general both the metam sodium alone and in combination with 1,3-D+C-17 were highly efficacious when compared to methyl bromide. Short polyethylene film retention times and short aeration times resulted in poor stands and poor crop vigor while relatively long polyethylene film retention times and long aeration periods at the same rates typically resulted in high stand counts and vigor. Combination treatments were more phytotoxic to germinating seed of tobacco and tomato. Vigor and stand counts of the seedlings were higher as aeration time increased, suggesting phytotoxic residues dissipate with time. Method of application of metam sodium, either injected with chisels or sprayed onto the soil surface and incorporated with a tractor-powered tiller alone or co- applied with 1,3-D+C-17 chisel injected, did not affect the efficacy of the treatments. Caution regarding phytotoxicity must be exercised when seeding into soil fumigated with metam sodium alone or combined with 1,3-D+C-17. Additional work will be required to establish safety periods required prior to transplanting crops into fumigated soil.     

Host serum proteins in Taenia saginata metacestode fluid

Taenia saginata cyst fluid was examined for host proteins; IgG1 and IgG2 as well as haemolytic complement activity were detected. Polyacrylamide gel electrophoresis revealed differences in proteinograms among the samples taken from 1-, 4-, and 10-month old cysts. Fluid from older cysts had fewer protein components and showed a weaker antigenic reaction with sera of bovines infected with T. saginata than that of younger cysts. The roles of antibody and complement in initiating degeneration of the parasite are discussed.     

Bleeding tendency as a cause of epistaxis in the dog

Based upon a survey of the pathophysiology of haemostasis, the causes of epistaxis in the dog are discussed, and guidelines for a diagnostic and therapeutic approach are given.     

Effect of surfactants on weight gain in mice

A study was conducted to determine if four surfactants can induce increased weight gain in the mouse. Basic-H, Triton X-100, Amway All Purpose Adjuvant and X-77 were put in water and fed to various groups of ICR 21 day old female mice for a period of 43 days. All the mice were clinically normal throughout the study period. Pathological examination of a random sample of the mice revealed no gross pathological changes. Similarly, histopathological examination of the lungs, livers and intestines did not reveal any visible lesions. Basic-H and Amway surfactants induced weight gain, though not significantly, better at 0.1% (V/V) concentration while X-77 and Triton X-100 induced weight gain better at 0.4% (V/V) concentration. Overall results show that none of the surfactants tested induced significant weight gain.     

Relationship between blood iron parameters and trichinellosis in swine

Reduced levels of total iron binding capacity and unsaturated iron binding capacity were observed in the blood of trichinous and iron-injected trichinous pigs. No change was observed in their serum iron and saturation concentration levels. Also, reduced iron concentration levels were observed in the livers of trichinous pigs, while increased iron concentration levels were observed in the spleens of trichinous pigs and the livers and spleens of iron-injected pigs. No difference was found with regard to weight gains, number of larvae per gram of tissues, or histologic characteristics of 'nurse cells'.