LDH and LDH Isoenzymes of Synovial Fluid in the Horse

LDH is an intracellular enzyme, which when cells degenerate is released to the extracellular spaces and body fluids. Cells and organs in the mammalian body differ from each other with respect to their LDH isoenzyme patterns. These circumstances have led to the use of LDH isoenzyme determinations in laboratory diagnostic work. In the present investigation total LDH activity and LDH isoenzyme distribution in equine synovial fluid from healthy joints, joints with serous arthritis, osteochondrosis dissecans and arthrosis, were determined. The fluids from the diseased joints differed from normal synovial fluid with respect to total LDH activity, and the different joint diseases each seemed to give rise to a characteristic isoenzyme pattern. In order to examine possible sources of the increased LDH activity and altered isoenzyme patterns, blood plasma, red and white blood cells, synovial membrane and articular cartilage were also studied. It was found that LDH4 and LDH5 were present in high amounts in articular cartilage, and an increase in these isoenzymes was the most characteristic feature in synovial fluid from joints with arthrosis. The results were discussed in view of possible diagnostic value of isoenzyme determinations on synovial fluid.     

Synovial fluid and plasma concentrations of ceftiofur after regional intravenous perfusion in the horse

OBJECTIVE: To determine radiocarpal (RC) joint synovial fluid and plasma ceftiofur concentrations after regional intravenous perfusion (RIP) and systemic intravenous (IV) administration. STUDY DESIGN: Experimental cross-over study. ANIMALS: Five normal adult horses. METHODS: One RC joint was randomly selected for RIP and the contralateral RC joint was sampled to determine intrasynovial ceftiofur concentrations after IV administration. Wash-out between IV and RIP was > or = 14 days. After surgical introduction of an intraarticular catheter, ceftiofur (2 g) was administered under general anesthesia either IV or by RIP after tourniquet application. Plasma and synovial fluid were collected over 24 hours. Samples were analyzed using high-performance liquid chromatography with ultraviolet detection and the results were statistically analyzed using a linear mixed effect model. RESULTS: Mean synovial fluid ceftiofur concentrations were consistently higher after RIP than after IV administration and were > 1 mug/mL (minimal inhibitory concentration [MIC] for common pathogens) for >24 hours. Mean synovial fluid peak concentration of ceftiofur after RIP and IV administration was 392.7+/-103.29 microg/mL at 0.5 hours postinjection (HPI) and 2.72+/-0.31 mug/mL at 1 HPI, respectively. Large variations in synovial fluid and plasma ceftiofur concentrations were observed between horses regardless of administration technique. RIP did not cause adverse effects. CONCLUSIONS: Under the present experimental conditions RIP with ceftiofur (2 g) induced significantly higher intraarticular antibiotic concentrations in the RC joint in comparison with IV administration. Moreover, after RIP, synovial fluid ceftiofur concentrations remain above the MIC for common pathogens (1 microg/mL) for > 24 hours. No adverse effects from the technique or the antibiotic were observed. CLINICAL RELEVANCE: RIP with high doses of ceftiofur may be a beneficial adjunctive therapy when treating equine synovial infections which are caused by cephalosporin susceptible microorganisms.     

Synovial membrane microarthroscopy of the equine midcarpal joint

OBJECTIVE: To evaluate the value of microarthroscopy in the equine midcarpal joint using the vital stains methylene blue, trypan blue, neutral red, and Janus green B to observe components of the synovial lamina propria, vascular architecture, and synoviocytes. STUDY DESIGN: Experimental. ANIMALS: Ten horses. METHODS: Microarthroscopy of left and right midcarpal joints was performed with and without vital staining of the synovium. Four vital stains (methylene blue, trypan blue, neutral red, and Janus green B) were evaluated, with each stain used in 5 joints. Synovial biopsy specimens were collected from the dorsomedial and dorsolateral aspects of the joint. RESULTS: All dyes were biocompatible. At x 60 without vital staining, synovial surface topography, vascular network, and translucency were observed. Intra-articular vital dyes improved evaluation of synovial surface topography. At x 150 with vital staining, individual synoviocytes were clearly identified with all dyes, except neutral red. Although methylene blue provided the best in vivo microscopic differentiation of the structure of the intima, trypan blue had superior retention in conventionally processed synovial biopsies. CONCLUSIONS: Methylene blue, trypan blue, neutral red, and Janus green B stains can be used safely for microarthroscopy. Good visualization of cells and vascular network can be obtained by microarthroscopy, and microarthroscopic evaluation of the synovium compares favorably with conventional histologic evaluation of biopsy specimens. CLINICAL RELEVANCE: Microarthroscopy may be beneficial in both research and clinical diagnosis of equine articular diseases.     

Methods and Errors in Measurements of Synovial Fluid Volume in Stifles with Low Volume and High Viscosity Synovial Fluid. An Experimental Study in Goats

Synovial fluid (SF) volume was calculated using various methods in the stifles of goats, in which the cranial cruciate ligament had been transected on one side. Measurements were performed prior to surgery and again 4,8, and 18 weeks following surgery, by measuring the dilution of an injected radioactive tracer diluted by the SF. Later, 7 months following surgery, SF volume measurements using simple arthrocentesis were performed on stifles in 9 of the goats, and the SF that could not be aspirated, was calculated using 2 indirect methods simultaneously on identical fluids in 3 of these goats. SF was also collected directly during staged arthrotomy of the stifles in 4 goats.There were conflicting results between methods, but the resulting calculated SF volumes seemed to be larger in the operated stifles compared to the controls for all the methods at about the same degree. The 2 indirect methods used to calculate the fluid remaining in the joints following arthrocentesis gave disparate volume calculations. The experiments revealed sources of error in all methods. Direct methods failed to acquire the total fluid volume, and indirect methods were subject to improper mixing and escape of the injected fluid or synovial fluid or both. It was concluded that none of the methods could be used to measure the “true” volume of SF, if such a concept exists and can be defined. None of the methods were considered reliable to compare volumes in different type of joints containing this type of fluid. It was, however, concluded that all the methods gave indication of increased SF volume present on a relative basis when paired joints were compared.     

奶牛正常腕,跗关节滑液主要生化成分的分析

采用常规实验室检查方法对６头中国荷斯坦奶牛腕关节和跗关节滑液中蛋白质、葡萄糖、钙和磷的含量进行了测定,并与血清中相应成分进行比较分析,结果,左右关节之间无差异,腕关节的总蛋白、白蛋白、球蛋白均比跗关节的低,但差异不显著。血清中的蛋白质、葡萄糖及磷的含量极显著地高于滑液中,只有钙的含量极显著地低于滑液中的含量。     

Determination of MMP-2 and -9 activities in synovial fluid of horses with osteoarthritic and arthritic joint diseases using gelatin zymography and immunocapture activity assays

REASON FOR PERFORMING STUDY: Matrix metalloproteinases (MMPs)-2 and -9 activities have been found elevated in synovial fluid from various joint diseases in man. However, in the horse few data are available. OBJECTIVES: To explore the clinical significance of MMP-2 and -9 activities in synovial fluid of horses with different forms of joint diseases. METHODS: Gelatin zymography and MMP-2 and -9 immunocapture activity assays were applied on synovial fluids from control joints and joints with aseptic joint disease (AJD) and septic arthritis (SA). Additionally, MMP-2 and -9 activities were measured in samples from SA to monitor the disease process. RESULTS: Zymographic analysis revealed that samples from AJD and SA contained significantly increased latent MMP-2 activity compared to controls. Samples from SA showed significantly increased monomeric latent MMP-9 activity compared with all other affected joints and controls. Trace amounts of MMP-9 activity, due to the active and dimer form, were detected in samples from SA; however, these bands were absent in samples from AJD and controls. Using immunocapture activity assays, MMP-2 and -9 activities were found to be significantly elevated in joints from SA compared to controls and AJD samples. MMP-2 activity in samples from AJD was significantly increased compared to controls. Both MMP activities decreased in the joints from SA in the course of successful therapy. CONCLUSIONS: Data from zymographic analysis confirmed that MMP-2 and -9 were elevated in equine joint diseases. Immunocapture activity assays have been shown to be suitable for the quantitative determination of MMP-2 and -9 activities in synovial fluid of horses. Both MMP-2 and -9 activities seem to be useful to indicate SA, and MMP-2 activity might be a suitable marker for AJD. POTENTIAL RELEVANCE: These findings encourage the potential use of MMP-2 and -9 as additional aids to clinical investigation. Further work is required to validate the clinical significance of MMP activities in the progress of different joint diseases in horses.     

Synovial osteochondromatosis

Synovial osteochondromatosis represents a very unusual synovial response in the horse and can be either primary or secondary. The condition describes metaplastic and focal formation of cartilage within the intimal layer of the synovial membrane. The condition is also uncommon in other domestic animals, although occurs more frequently in man. Treatment usually involves arthroscopic removal of osteochondral bodies and resection of abnormal synovium. Recurrence is quite common and malignant transformation rarely occurs although, to date, this has not been reported in the horse.     

马关节滑液、血清乳酸脱氢酶及其同工酶和它们在疾病状态下的变化

用king氏法和聚丙烯酰胺凝胶电泳法测得马关节滑液和血清乳酸脱氢酶(LDH)及其同工酶正常值。滑液和血清5种LDH同工酶电泳迁移率相同。各种LDH同工酶的分子量和热稳定性均有明显差异。抗尿素作用,血清和滑液LDH同工酶基本相似。Mg~(++)是LDH激活剂,最佳激活浓度为10~(-3)M;Zn~(++)、Ca~(++)、Cu~(++)、Hg~(++)、谷胱甘肽、半胱氨酸、碘乙酸、EDTA等为LDH抑制剂。在四肢的急性或亚急性关节和骨疾病时,滑液LDH明显升高,其同工酶谱随疾病发生部位不同而不同,并与血清一致,但慢性关节疾病时LDH及其同工酶变化则不一致。     

Light and Electron Microscopic Investigation of Equine Synovial Membrane: A Comparison Between Healthy Joints and joints with Intraarticular Fractures and Osteochondrosis Dissecans

Light and electron microscopic examination was made on equine synovial membrane from 23 healthy joints, nine joints with synovitis caused by intraarticular fracture and 10 joints with synovitis caused by osteochondrosis dissecans. Histologically as well as ultrastructurally the equine synovial membrane from healthy joints was of principally the same character as described in other species. Three types of synovial membrane — areolar, fibrous and adipose — and two types of lining cell were distinguished histologically. Ultrastructurally three types of lining cells were distinguished: A and Β type and an intermediate cell type. In healthy joints they were loosely arranged, parallel to the joint surface in an intercellular matrix, which was in direct continuity with the joint space. In joints with intraarticular fracture there was mild inflammation of the synovial membrane. There was elongation and hyperplasia of the lining cells with a relative increase in type A cells. The cell surface of lining cells was increased through filopodia. There was also an increase in cytoplasmic organelles i.e. hyperplasia of rough endoplasmic reticulum and Golgi complexes in Β type cells and an increase in lysosomes, and increased numbers of vesicles of varying types in A cells. In joints with osteochondrosis dissecans the lining cell hyperplasia and the inflammation in the synovial membrane were more prominent. Ultrastructurally the same alterations as in the previous group were seen including a relative increase in the number of A cells but degenerative changes were common in the lining cells. These changes were dilatation and vesiculation of rough endoplasmic reticulum, mitochondrial condensation, dilatation of the nuclear envelope and loss of plasma membranes, leading to disintegration of cells.     

Oestrogens in New Zealand pasture plants

Extract

The following case history is reported as suggesting the contra-indication of androgenic therapy in such cases.     

Characteristics of Normal Equine Tarsal Synovial Fluid

Physical, biochemical, and cytologic properties of synovial fluid from normal equine tarsal joints were investigated. Tarsal synovial fluid was pale yellow, clear, free of flocculent material, and did not clot. Volume varied in direct proportion to individual tarsal joint size. Relative viscosity was related to volume, polymerization and quantity of hyaluronic acid, and protein concentration. Mucinous precipitate quality (hyaluronic acid polymerization) was uniformly high.

Results of certain analyses of serum were compared with those of tarsal synovial fluid. Tarsal synovial fluid protein concentration was low in conjunction with a high A:G ratio. Serum: synovial fluid sugar ratio was 1.24:1. Serum ALP, ACP, LDH, GOT, and GPT activity levels were higher than their corresponding levels of activity in tarsal synovial fluid. Serum ALD activity level was slightly lower than its tarsal synovial fluid counterpart. Total erythrocyte counts ranged markedly, while total leukocyte counts were uniform and low. Lymphocytes were the predominant synovial fluid cell type.

     

Synovial fluid chondroitin sulphate indicates abnormal joint metabolism in asymptomatic osteochondritic horses

Reasons for performing study: Alternative methods to evaluate the joint condition in asymptomatic osteochondrosis dissecans (OCD) and other joint diseases may be useful. Objectives: To investigate possible changes in synovial fluid composition that may lead to joint conditions in asymptomatic OCD, in mature horses. Methods: Animals aged >2 years, of different breeds, with OCD in the intermediate ridge of distal tibia, symptomatic or not, were studied. Synovial fluid samples (10 healthy; 11 asymptomatic OCD; 25 symptomatic OCD) were collected by arthroscopy from 29 horses. Glycosaminoglycans (GAGs) were analysed by a combination of agarose gel electrophoresis and enzymatic degradation with specific GAG lyases. The viscosity, white blood cell (WBC) count, protein concentration and hyaluronic acid (HA) molecular weight were also determined. Results: The method used here to analyse synovial fluid GAGs is reliable, reproducible and specific. The main synovial fluid GAGs are HA and chondroitin sulphate (CS), 93% and 7% respectively in normal horses. In symptomatic OCD, the concentrations of both increased (expressed as GAG/urea ratios), but CS increased more. The CS increased also in asymptomatic OCD. An inflammatory reaction was suggested by the increased WBC counts in OCD. The molecular weight of the synovial fluid HA was reduced in OCD, explaining the lower viscosity observed. Conclusions: The increased CS in synovial fluid of OCD joints in mature horses suggests that the synovial fluid CS and the WBC count are good markers of the joint conditions, allowing the identification of pathological phase in joint diseases. Potential relevance: The analysis of synovial fluid GAGs shows that cartilage damage occurs even in asymptomatic OCD, implying that arthroscopic removal of osteochondral fragments should be performed even in asymptomatic OCD.     

Blood Serum and Synovial Fluid in Bovine Laminitis and Arthritis,with Particular Reference to the Protein Composition

In 74 cases of clinical laminitis and in 10 cases of clinical discrete arthritis in adult cattle, the protein picture was changed in blood and synovial fluid. The cases of laminitis were clinically divided into one group without and another with general affection. The changes in serum γ- and α-globulins indicated a predominantly chronic inflammatory process in the arthritis groups and a more acute process in the laminitis groups. In discrete arthritis, the leukocyte concentration and all protein levels except the relative albumin concentration were increased. In laminitis without general affection no significant changes were found in synovial fluid, while the synovial protein pattern in laminitis with general affection indicated a simultaneous occurrence of arthritis. The combination of laminitis and arthritis might be an expression of different inflammatory reactions to common aetiological, still unidentified factors, indicating the existence of a disease complex which may manifest itself in hoof corium, in joints or in both.     

Synovial fluid eosinophilia: a case report in a dog and review of the literature

The first known report of synovial fluid eosinophilia in a dog is described here. The occurrence of eosinophils in joint fluid is rare. Sporadic cases have been recorded in humans and most can be related to immunemediated reactions, both parasitic and non- parasitic. The dog in this case report had 20-52% eosinophils in multiple joints, as well as hemarthrosis and marked mononuclear cell reactivity. An intense peripheral eosinophilia was demonstrated one week later. The associated lameness resolved with non-steroidal anti-inflammatory therapy. Lack of remarkable history or other clinical symptoms led to a diagnosis of idiopathic, eosinophilic, polyarthritis, likely immune-mediated.     

Physiological Concentrations of Acute-Phase Proteins and Immunoglobulins in Equine Synovial Fluid

Synovial fluid (SF) is capable of reflecting infectious, immunological, or inflammatory joint conditions in horses by altering its composition and appearance. Although plasma and SF compositions are quantitatively different, this latter compartment reflects changes in plasma macromolecules. Therefore, changes in serum immunoglobulin protein concentrations tend also to alter intracapsular levels. Therefore, it is necessary to know the physiological concentrations of proteins present in SF. The aim of this study was to determine the levels of total protein, albumin, transferrin, haptoglobin, α1-acid glycoprotein, ceruloplasmin, and immunoglobulins A and G in SF of six healthy horses. The synovial proteinogram was obtained by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The SF proteins reached a maximum of 25% of serum concentrations, varying inversely with molecular weight of the protein, except for the ceruloplasmin.