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- Author or Editor: Stacy A. Semevolos x
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Abstract
Objective—To determine molecular changes in the expression of insulin-like growth factor-I (IGF-I) and transforming growth factor-β1 (TGF-β1) in horses with osteochondrosis, and to characterize expression of matrix aggrecan and collagen types I, II, and X in articular cartilage of affected joints.
Sample Population—Articular cartilage from affected stifle or shoulder joints of 11 horses with naturally acquired osteochondrosis and corresponding joints of 11 clinically normal horses.
Procedure—Harvested specimens were snap frozen in liquid nitrogen, and total RNA was isolated. Specimens were fixed in 4% paraformaldehyde for histologic examinations. Expression of matrix molecules was assessed by analysis of northern blots and in situ hybridization, using equine-specific cDNA probes and riboprobes, respectively. Expression of IGF-I and TGF-β1 was assessed by use of noncompetitive quantitative polymerase chain reaction, in situ hybridization, and immunohistochemical analysis.
Results—Cartilage obtained from osteochondrosis lesions had significantly greater expression of IGF-I, compared with normal cartilage. Expression of TGF- β1 and collagen type I were higher, but not significantly so, in affected tissues. Expression of aggrecan or collagen types II and X did not differ between affected and clinically normal cartilage.
Conclusions and Clinical Relevance—Increased expression of growth factors and collagen type I was found in cartilage from osteochondrosis lesions. However, this probably reflects a healing response to injured tissue rather than a primary alteration. Therefore, methods aimed at altering concentrations of growth factors in cartilage of growing horses would be unlikely to alter the incidence or progress of the disease. (Am J Vet Res 2001;62:1088–1094)
Abstract
Objective—To determine the anatomic communications among compartments within the carpus, metacarpophalangeal and metatarsophalangeal joints, stifle joint, and tarsus in llamas.
Sample Population—88 limbs from 22 llamas necropsied because of reasons unrelated to disease of the carpus; tarsus; or metacarpophalangeal, metatarsophalangeal, or stifle joints.
Procedure—1 compartment (randomly assigned) of each joint was injected with blue latex solution. Communication between joint compartments was determined by observation of latex in adjacent compartments following frozen sectioning.
Results—Of the 44 carpi, 30 (68%) had anatomic separation between the radiocarpal and middle carpal joints, whereas the remaining 14 (32%) had communication between the radiocarpal and middle carpal joints. In the metacarpophalangeal or metatarsophalangeal joints, medial and lateral joint compartments remained separate in 83 of 88 (94%) joints injected. The tibiotarsal and proximal intertarsal joints communicated in all tarsi examined, whereas 14 of 38 (37%) communicated between the proximal intertarsal and distal intertarsal joints. Communication between the distal intertarsal and tarsometatarsal joints was detected in 17 of 25 (68%) specimens; all 4 tarsal joints communicated in 11 of 42 (26%) specimens examined. Examination of 33 stifle joints that were successfully injected revealed communication between the femoropatellar, medial femorotibial, and lateral femorotibial joints.
Conclusions and Clinical Relevance—These data suggest that it is important to determine the joint communications specific to each llama prior to treatment of septic arthritis. The metacarpophalangeal or metatarsophalangeal joint compartments may be considered separate, although the lateral and medial compartments infrequently communicate along the proximal palmar or plantar aspect. (Am J Vet Res 2005;66:1437–1440)
Abstract
Objective—To characterize serum trace mineral, sex steroid hormone, and vitamin D concentrations and identify factors associated with metacarpophalangeal and metatarsophalangeal hyperextension in llamas and alpacas.
Samples—Serum samples from 79 llamas and 15 alpacas and owner survey data for 573 llamas and 399 alpacas.
Procedures—Serum samples were stored at −20°C until analysis and were evaluated for trace mineral, vitamin D, estradiol, progesterone, and testosterone concentrations. Information regarding age of onset, number of affected animals in herd, feed and supplements given, type of housing, and management practices was obtained in an owner survey.
Results—Higher serum zinc and iron concentrations were associated with metacarpophalangeal and metatarsophalangeal hyperextension in camelids, compared with controls. In summer and fall months, vitamin D concentrations were significantly higher in affected camelids than controls. Overall prevalence was 13.3% in llamas, compared with 0.7% in alpacas. No management factors were found to be predictive of this condition. No other factors examined were associated with metacarpophalangeal and metatarsophalangeal hyperextension.
Conclusions and Clinical Relevance—Despite similar supplementation practices and environmental conditions between affected and unaffected animals, an association of high serum zinc, iron, and vitamin D concentrations in affected camelids, compared with controls, may indicate differences of intake or absorption of dietary supplements.
Abstract
OBJECTIVE To evaluate 2 processing methods (commercial kit vs conical tube centrifugation) for preparing platelet rich plasma (PRP) for use in llamas and alpacas.
SAMPLES Blood samples (30 mL each) aseptically collected from 6 healthy llamas and 6 healthy alpacas.
PROCEDURES PRP was prepared from blood samples by use of a commercial kit and by double-step conical tube centrifugation. A CBC was performed for blood and PRP samples. Platelets in PRP samples were activated by means of a freeze-thaw method with or without 23mM CaCl2, and concentrations of platelet-derived growth factor-BB and transforming growth factor-β1 were measured. Values were compared between processing methods and camelid species.
RESULTS Blood CBC values for llamas and alpacas were similar. The commercial kit yielded a significantly greater degree of platelet enrichment (mean increase, 8.5 fold vs 2.8 fold) and WBC enrichment (mean increase, 3.7 fold vs 1.9 fold) than did conical tube centrifugation. Llamas had a significantly greater degree of platelet enrichment than alpacas by either processing method. No difference in WBC enrichment was identified between species. Concentrations of both growth factors were significantly greater in PRP samples obtained by use of the commercial kit versus those obtained by conical tube centrifugation.
CONCLUSIONS AND CLINICAL RELEVANCE For blood samples from camelids, the commercial kit yielded a PRP product with a higher platelet and WBC concentration than achieved by conical tube centrifugation. Optimal PRP platelet and WBC concentrations for various applications need to be determined for llamas and alpacas.
Abstract
Objective—To determine the mRNA expression of bone morphogenetic protein (BMP)-6 and -2 and a BMP antagonist (Noggin) in horses with osteochondrosis.
Sample Population—Samples of articular cartilage from affected stifle or shoulder joints of 10 immature horses with naturally acquired osteochondrosis and corresponding joints of 9 clinically normal horses of similar age; additionally, samples of distal femoral growth plate cartilage and distal femoral articular cartilage were obtained from a normal equine fetus.
Procedure—Cartilage specimens were snap-frozen in liquid nitrogen, and total RNA was isolated. Adjacent specimens were fixed in 4% paraformaldehyde for histologic examination. Expression of BMP-6, BMP-2, and Noggin mRNA was evaluated by real-time quantitative polymerase chain reaction (PCR) assays. Spatial tissue mRNA expression of BMP-6 was determined by in situ hybridization.
Results—Nucleotide sequences were obtained for portions of the BMP-6 propeptide and mature peptide region, as well as the signal and mature peptide region of Noggin. Expression of BMP-6, BMP-2, and Noggin mRNA was found to be similar in cartilage from normal and osteochondrosis-affected horses. Spatial expression of BMP-6 correlated with the middle and deep layers of articular cartilage; no differences were observed in overall expression between cartilage specimens from the 2 groups of horses. No expression of BMP-6 was found in the superficial layer, subchondral bone, or osteochondrosis-affected cleft fibrous tissue.
Conclusions and Clinical Relevance—Although these signaling peptides may play important roles in cartilage differentiation, results did not provide evidence to suggest that they are involved in the disease process of osteochondrosis. (Am J Vet Res 2004;65:110–115)
Abstract
OBJECTIVE To determine and compare mean standing extension and maximum flexion angles of various joints in healthy adult alpacas and llamas, and determine the reliability of goniometric data within and between 2 observers for each joint of interest.
SAMPLE 6 healthy adult llamas and 6 healthy adult alpacas.
PROCEDURES The shoulder joint, elbow joint, carpal, and metacarpophalangeal (MCP) joints of the forelimbs and the hip joint, stifle joint, tarsal, and metatarsophalangeal (MTP) joints of the hind limbs were investigated. Each articulation was measured with a universal goniometer by 2 observers, who each obtained 2 measurements when each joint was maintained in standing extension and in maximal passive flexion. Two sample (unpaired) t tests were performed for comparisons of mean standing extension and maximum passive flexion angles between alpacas and llamas. Intraclass correlation coefficients were calculated for each articulation to assess interobserver and intra-observer reliability of measurements.
RESULTS Llamas had larger mean standing extension angles than alpacas for the tarsal and elbow joint, but there were no significant differences between species for all other joints. For all joints, flexion measurements did not differ significantly between the 2 species. For most joints, the reliability of goniometric data between observers was good to excellent (intraclass correlation coefficients, 0.6 to 0.95)
CONCLUSIONS AND CLINICAL RELEVANCE Except for the elbow joint and tarsus in extension, the angle of limb articulations during flexion and extension can be considered similar for alpacas and llamas. These measurements have relevance for veterinary surgeons when assessing joint mobility and conformation and determining appropriate angles for arthrodesis.
Abstract
Objective—To determine the molecular and histologic characteristics of hyperextension of the metacarpophalangeal and metatarsophalangeal joints in adult llamas.
Animals—12 adult llamas (6 with bilateral hyperextension of the metacarpophalangeal or metatarsophalangeal joints [affected] and 6 age- and sex-matched clinically normal control llamas).
Procedures—Llamas were euthanized, and specimens of superficial digital flexor tendon, deep digital flexor tendon, and suspensory ligament were obtained from 4 areas and snap frozen in liquid nitrogen or suspended in neutral-buffered 10% formalin. Histologic evaluation of collagen fiber orientation, elastin content, and proteoglycan content was performed by use of Masson trichrome, picrosirius red, Verhoeff, and Alcian blue stains. Total RNA was isolated from frozen suspensory ligament specimens. Gene expression of collagen types I and III, lysyl oxidase, and matrix metalloproteinase-13 was evaluated with a real-time quantitative reverse transcriptase PCR assay.
Results—Gene expression of collagen types I and III, lysyl oxidase, and matrix metalloproteinase-13 in suspensory ligaments was similar between affected and control llamas. Collagen orientation and elastin content of the flexor tendons and suspensory ligaments were also similar between the groups. Proteoglycan content was low in most specimens but was focally increased in discrete lesions of suspensory ligaments in 2 affected and 2 control llamas.
Conclusions and Clinical Relevance—Hyperextension of the metacarpophalangeal or metatarsophalangeal joints in llamas did not appear to be caused by degeneration or inflammation of the suspensory ligament. Although focal proteoglycan accumulation existed in the suspensory ligaments of 2 affected llamas, widespread abnormal connective tissue proteoglycan accumulation was not found.
Abstract
Objective—To evaluate molecular and histologic characteristics of the superficial digital flexor tendon (SDFT), deep digital flexor tendon (DDFT), and suspensory ligament (SL) and assess trace-mineral concentrations in serum, liver, and hair of juvenile llamas with metacarpophalangeal and metatarsophalangeal joint hyperextension.
Animals—12 juvenile llamas (6 with bilateral hyperextension of metacarpophalangeal joints, metatarsophalangeal joints, or both and 6 clinically normal control llamas).
Procedures—Radiography and ultrasonography of metacarpophalangeal and metatarsophalangeal regions were performed. Llamas were euthanized, and SDFT, DDFT, and SL samples were collected for histologic evaluation of collagen and elastin content and orientation, proteoglycan content, and collagen type III immunohistochemistry. Total RNA was isolated from SL tissue, and gene expression of collagen types I and III, lysyl oxidase, and matrix metalloproteinase-13 was evaluated via real-time quantitative reverse transcriptase PCR assay. Liver, serum, and hair samples were evaluated for trace mineral content.
Results—Collagen type III gene expression and proteoglycan content were significantly increased in SL samples of affected juvenile llamas, compared with those of control llamas. No difference was detected in collagen and elastin content and orientation or in gene expression of collagen type I, lysyl oxidase, or matrix metalloproteinase-13 between groups. Affected llamas had significantly increased serum molybdenum and decreased liver cobalt concentrations, compared with values for control llamas.
Conclusions and Clinical Relevance—Increased collagen type III gene expression and proteoglycan content in SL samples of affected juvenile llamas provided evidence of ongoing SL matrix repair. Trace mineral differences may have been attributable to dietary imbalances in affected llamas.
Abstract
Objective—To determine the morphologic and biochemical characteristics of hyperextension of the metacarpophalangeal and metatarsophalangeal joints in llamas.
Animals—12 adult llamas (6 with bilateral hyperextension of the metacarpophalangeal or metatarsophalangeal joints and 6 age- and sex-matched control llamas).
Procedures—Llamas were evaluated by use of lameness examination, ultrasonography, and radiography. A CBC, serum biochemical analysis, and determination of concentrations of trace minerals in serum and liver samples were performed. Llamas were euthanized, and samples of the superficial digital flexor tendon, deep digital flexor tendon, and suspensory ligament were obtained from 4 areas and snap-frozen in liquid nitrogen or suspended in neutral-buffered 10% formalin. Immunohistochemical evaluation of collagen types I and III and assays for measurement of lysyl oxidase activity were performed.
Results—2 affected llamas had a visible gait deficit associated with metacarpophalangeal joint hyperextension. Radiographic evidence of osteoarthritis was detected in 1 severely affected llama, and ultrasonographic changes of soft tissue mineralization and suspensory desmitis were observed in 2 llamas. Liver concentrations of copper were lower and serum concentrations of zinc higher in affected llamas, compared with values in control llamas. Lysyl oxidase activity and collagen distribution did not differ significantly between groups.
Conclusions and Clinical Relevance—Hyperextension of the metacarpophalangeal or metatarsophalangeal joints in llamas does not appear to be the result of injury or degeneration of the suspensory ligament or flexor tendons. Lower copper concentrations coupled with higher zinc concentrations in affected llamas may be indicative of secondary copper deficiency.
Abstract
Objective—To compare postoperative complications, short- and long-term survival, and surgical times for hand-sewn end-to-end (EE), stapled functional end-toend (FEE), and stapled side-to-side (SS) anastomotic techniques for jejunal resection in horses.
Design—Retrospective study.
Animals—59 horses.
Procedure—Medical records were reviewed to obtain signalment, diagnosis, treatment, and outcome for horses that underwent jejunojejunostomy in our hospital. Only horses that recovered from anesthesia were included in the study.
Results—Among the 59 horses, there were 33 EE, 15 FEE, and 11 SS anastomoses. No difference was found in duration of surgery among the 3 techniques. The most common postoperative complications were colic episodes (56%), ileus (53%), diarrhea (20%), and adhesions (15%). Horses with SS anastomosis had a significantly shorter duration of postoperative ileus than the EE group did. No significant difference in duration of postoperative ileus was found among the other groups. No difference was found among the 3 anastomotic techniques in regard to survival rate at the time of discharge, 6 months after surgery, or 1 year after surgery. Overall survival rates after jejunal anastomosis were 88% at the time of discharge, 65% at 6 months after surgery, and 57% at ≥ 1 year after surgery.
Conclusions and Clinical Relevance—The handsewn EE, stapled FEE, and stapled SS anastomotic techniques should be considered equivalent methods for small intestinal anastomosis in the horse. However, the stapled SS technique may be preferred because of possible decreased duration of postoperative ileus. (J Am Vet Med Assoc 2002;220:215–218)