Detection of osteoarthritis in dogs by metabolic, pro-inflammatory and degenerative synovial fluid biomarkers and traditional radiographic screening: A pilot study.

Secondary osteoarthritis (OA) is a slow progressive, common disorder of synovial joints in dogs. It is characterized by a loss of balance between the synthesis and degeneration of articular cartilage components. Its diagnosis is currently based on the presence of clear radiographic changes, which only occur in the later stages of the disease. Hence, early diagnosis of OA remains a major problem. Therefore, interest in synovial fluid (SF) biomarkers has emerged. Besides pro-inflammatory and degenerative markers, i.e. tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), tenascin-c (TN-C) and matrix metalloproteinase-2 (MMP-2), metabolic parameters, i.e. pH, glucose and lactate, can potentially be used to detect OA. The current study demonstrated statistically significant differences in the SF levels of pH, glucose and lactate between OA-affected and normal joints. In addition, the in-house validated immuno-assays for TNF-alpha, IL-1beta, TN-C and MMP-2 allowed to demonstrate also statistically significant differences in the SF concentrations for all these biomarkers - except TNF-alpha - between OA-affected and normal joints. However, no correlation was found between any of these biomarkers and the currently used radiographic scoring system for OA in dogs. Future research is warranted to explore the potential of these biomarkers in the early detection of OA and in the severity characterization of this disease.

The Histological Composition of Capsular Contracture Focussed on the Inner Layer of the Capsule: An Intra-Donor Baker-I Versus Baker-IV Comparison.

BACKGROUND: Capsular contracture remains one of the major complications after breast implantation surgery. The extent of capsular contraction is scored using the Baker scale. The aim of this study was to compare intra-individual Baker-I with Baker-IV capsules, and in particular the prevalence and histological properties of the inner capsule layer. METHODS: Twenty capsules from ten patients were included after bilateral explantation surgery due to unilateral capsular contracture (Baker-IV) after cosmetic augmentation with textured implants. All capsules underwent (immune-)histochemical analysis: haematoxylin-eosin (morphology), CD68 (macrophages), cytokeratin (epithelial cells) and vimentin (fibroblasts), and were visually scored for cell density and the presence of an inner layer and measured for thickness. RESULTS: Baker-IV (n = 10) capsules were significantly thicker compared to Baker-I (n = 10) capsules (P = 0.004). An inner layer was present in 8 Baker-I capsules. All Baker-I capsules were vimentin and CD68-positive and cytokeratin-negative. Positive vimentin was seen throughout the inner layer, and CD-68 staining was observed adjacent to the intermediate capsule layer. In contrast, only 2 Baker-IV capsules had an inner layer, of which only 1 showed the same profile as Baker-I capsules (P = 0.016). No cytokeratin positivity was seen in any capsule. In Baker-IV capsules, outer layers showed more positivity for both vimentin and CD68. CONCLUSION: The inner layer is morphologically consistent with synovial metaplasia and is more prevalent in healthy, uncontracted Baker-I capsules. This inverse relation between the presence of the inner layer and higher Baker classification or pathological contracture could indicate a protective role of the inner layer against capsular contracture formation. LEVEL OF EVIDENCE III: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Primary and concomitant flexor enthesopathy of the canine elbow.

OBJECTIVES: To report the characteristics of two types of flexor enthesopathy, primary and concomitant, based on different diagnostic techniques. MATERIALS AND METHODS: Over a period of three years a prospective study was performed on dogs admitted for the complaint of elbow lameness. Based on the radiographic findings a selection of dogs underwent a complete series of different imaging modalities. With each technique, pathology of the medial epicondyle and the presence of other elbow disorders were recorded. All joints with signs of flexor pathology apparent with at least three techniques were selected. A distinction was made between primary and concomitant flexor enthesopathy based on the absence or presence of other elbow disorders. RESULTS: Primary flexor enthesopathy was diagnosed in 23 joints and concomitant flexor enthesopathy in 20 joints. In 43% of the joints with primary and in 75% of the joints with concomitant flexor enthesopathy, pathology at the medial epicondyle was demonstrated by all techniques. All joints with concomitant flexor enthesopathy had a diagnosis of medial coronoid disease, osteochondritis dissecans, or both. CLINICAL SIGNIFICANCE: Pathology at the medial epicondyle is a sign of flexor enthesopathy. It may be present as the only sign in a joint with primary flexor enthesopathy or concomitant with other elbow pathology. In both groups flexor lesions can be demonstrated with different imaging techniques. The distinction between the primary and concomitant form is based on the presence or absence of other elbow pathology, mainly medial coronoid disease. Recognizing both types is important for a correct treatment decision.

The use of planar bone scintigraphy and HiSPECT for diagnosis of primary and concomitant flexor enthesopathy in the canine elbow.

OBJECTIVE: To investigate the possibilities and limitations of planar bone scintigraphy and high resolution single photon emission computed tomography (HiSPECT) to diagnose flexor enthesopathy and to distinguish primary flexor enthesopathy from the concomitant form. MATERIALS AND METHODS: A prospective study of 46 dogs with primary flexor enthesopathy, concomitant flexor enthesopathy, medial coronoid disease, and normal elbows was performed. All dogs underwent planar bone scintigraphy and HiSPECT imaging. The obtained images were visually scored for increased radiopharmaceutical uptake in the medial humeral epicondylar and medial coronoid process region using a score from 1-3. RESULTS: Planar bone scintigraphy demonstrated increased radiopharmaceutical uptake in all diseased elbow joints, except for one. HiSPECT demonstrated increased radiopharmaceutical uptake of the medial humeral epicondyle in nearly all clinically affected joints with primary and concomitant flexor enthesopathy. Additional uptake of the medial coronoid process was recorded in all clinically affected joints with concomitant flexor enthesopathy and in six out of 18 with primary flexor enthesopathy. No difference in intensity of the uptake was noticed. CLINICAL SIGNIFICANCE: Planar bone scintigraphy allows the attribution of lameness to the elbow joint in cases of primary flexor enthesopathy with minimal or even absent radiographic changes. The more detailed HiSPECT enables the localization of pathology within the elbow joint and is a sensitive technique to detect flexor enthesopathy. However HiSPECT is insufficient to distinguish primary from concomitant flexor enthesopathy.

Arthroscopic features of primary and concomitant flexor enthesopathy in the canine elbow.

OBJECTIVES: To investigate the possibilities and limitations of arthroscopy to detect flexor enthesopathy in dogs and to distinguish the primary from the concomitant form. MATERIALS AND METHODS: Fifty dogs (n = 94 elbow joints) were prospectively studied: dogs with primary flexor enthesopathy (n = 29), concomitant flexor enthesopathy (n = 36), elbow dysplasia (n = 18), and normal elbow joints (n = 11). All dogs underwent an arthroscopic examination of one or both elbow joints. Presence or absence of arthroscopic characteristics of flexor enthesopathy and of other elbow disorders were recorded. RESULTS: With arthroscopy, several pathological changes of the enthesis were observed in 100% of the joints of both flexor enthesopathy groups, but also in 72% of the joints with elbow dysplasia and 25% of the normal joints. No clear differences were seen between both flexor enthesopathy groups. CLINICAL SIGNIFICANCE: Arthroscopy allows a sensitive detection of flexor enthesopathy characteristics, although it is not very specific as these characteristics may also be found in joints without flexor enthesopathy. The similar aspect of both forms of flexor enthesopathy and the presence of mild irregularities at the medial coronoid process in joints with primary flexor enthesopathy impedes the arthroscopic differentiation between primary and concomitant forms, requiring additional diagnostic techniques to ensure a correct diagnosis.

The use of the HiSPECT bone scan in canine flexor enthesopathy and coronoid pathology.

In this study the use of the high resolution Micro-Single Photon Emission Tomography (HiSPECT) system with a radioactive bonemarker, (99m)Tc-oxidronate, was evaluated in dogs with coronoid pathology and/or flexor enthesopathy. Sixty-five elbows of 34 dogs were included. CT and HiSPECT were performed on all elbows, arthroscopy on 59. Tracer uptake in 8 anatomical regions was graded according to two models. Increased activity in the medial epicondylar region was associated with flexor pathology on CT (P=0.0002) and arthroscopy (P<0.0001) and increased uptake in the medial coronoid (P<0.0001) and the medial condylar area (P<0.013) with coronoid pathology. Uptake in the remaining areas was not associated with both pathologies. In conclusion, the improved resolution of the HiSPECT system allows identification of increased tracer uptake in the anatomical regions involved in coronoid pathology and flexor enthesopathy. This modality may improve the diagnostic potential of the bone scan in canine elbow disease.

Arthroscopic treatment of fragmented coronoid process with severe elbow incongruity. Long-term follow-up in eight Bernese Mountain Dogs.

The purpose of this study was to investigate the long-term treatment results of fragmented coronoid process (FCP) in joints with a radio-ulnar step greater than 3 mm. Treatment of these patients only consisted of fragment removal, without correction of the incongruity. The eight Bernese Mountain Dogs (11 joints) included in this study showed obvious clinical signs of elbow disease and were diagnosed with severe elbow incongruity and concomitant FCP in the time period from 1999-2003. At that time, elbow radiography, computed tomography, and arthroscopy were performed. The mean follow-up period was 5.6 years. The follow-up consisted of a telephone questionnaire combined with a clinical and radiographic re-evaluation at our clinic. The questionnaire revealed that all dogs were either free of lameness or only lame following heavy exercise. One dog sporadically required medication after heavy exercise. The owner satisfaction rate was 100%. The clinical re-evaluation did not reveal any signs of pain or lameness in all cases. Range-of- motion was decreased in nine of the 11 elbows. Radiographs revealed an increase in severity of osteoarthritis in every case.In this case series, arthroscopic fragment removal without treatment of incongruity was demonstrated to be a valuable treatment option and may provide a satisfactory long-term outcome.

Radiographic findings of the medial humeral epicondyle in 200 canine elbow joints.

OBJECTIVES: To determine the frequency and radiographic aspect of medial humeral epicondylar lesions as a primary or concomitant finding and to evaluate the association with osteoarthritis. METHODS: Medical records of dogs diagnosed with elbow lameness were reviewed. Inclusion criteria for this study were a complete clinical examination, a complete set of digital radiographs and a final diagnosis made by computed tomography or magnetic resonance imaging and arthroscopy. Changes of the medial humeral epicondyle were recorded and correlated with the radiographic osteoarthritis and final diagnosis. RESULTS: Eighty of the 200 elbows showed changes of the medial humeral epicondyle. In 12 of these 80 elbows, changes of the medial epicondyle were the only findings within the joint, and these elbows were diagnosed with primary flexor enthesopathy. In the remaining 68 elbows, other concomitant elbow pathologies were found. In those cases of concomitant epicondylar changes, high grades of osteoarthritis were recorded, while most elbows with primary flexor enthesopathy showed a low grade of osteoarthritis. CLINICAL SIGNIFICANCE: Changes of the medial humeral epicondyle are often considered clinically unimportant and are regarded as an expression of osteoarthritis. This study showed the relatively frequent presence of epicondylar changes of which the majority were considered concomitant to a primary elbow problem. If changes of the medial humeral epicondyle are the only pathologic finding (primary flexor enthesopathy) they should be considered as the cause of lameness and not as a sign of osteoarthritis.

Primary flexor enthesopathy of the canine elbow: imaging and arthroscopic findings in eight dogs with discrete radiographic changes.

OBJECTIVE: To describe the radiographic, ultrasonographic, computed tomography (CT), magnetic resonance imaging (MRI), and arthroscopic findings in eight dogs with elbow lameness caused by primary flexor enthesopathy. STUDY DESIGN: Clinical study. ANIMALS: Eight client-owned dogs. METHODS: In all dogs, lameness was localized to the elbow by clinical examination. Radiographic examination, ultrasound, CT and MRI were performed prior to arthroscopy. In seven dogs, surgical treatment and subsequent histopathology were performed. RESULTS: Primary enthesopathy of the medial epicondyle was diagnosed in eight dogs (13 joints) by combining the minimal radiographic changes with specific ultrasonographic, CT, MRI and arthroscopic findings at the medial epicondyle. In all joints, any other pathology could be excluded. Histopathology of the affected tissue revealed degeneration and metaplasia in the flexor muscles. CONCLUSIONS: Primary flexor enthesopathy at the medial epicondyle is an unrecognized condition and is a possible cause of elbow lameness in the dog. Diagnosis is based on specific imaging and arthroscopic findings. CLINICAL RELEVANCE: The most important cause of elbow lameness in dogs is medial coronoid disease. Often this condition presents with minimal radiographic and arthroscopic changes. In these cases, primary enthesopathy of the medial epicondyle should be considered as a differential diagnosis, in order to make the correct treatment decision.

Medial humeral epicondylar lesions in the canine elbow. A review of the literature.

A calcified fragment near the medial epicondyle of the humerus was originally described as an ununited medial epicondyle in 1966. Since then several papers reported similar lesions as a cause of elbow lameness. The aetiology and clinical significance of those lesions is poorly known. This paper gives an overview of the veterinary and human literature in an attempt to explain the aetiology and to suggest a diagnostic protocol and treatment plan.

Pharmacological effects of a specific leukotriene B(4) receptor antagonist (VML 295) on blood leukocytes, cutaneous inflammation and epidermal proliferation.

VML 295 (LY 293111) is a potent and specific leukotriene(4) receptor antagonist. It has previously been shown in human volunteers that VML 295 at a dosage of 48 mg twice daily inhibits the ex vivo leukotriene B(4) (LTB(4))-induced upregulation of CD11b on peripheral blood neutrophils. A clear dose-response relatinship was shown. In addition, VML 295 inhibits various inflammatory aspects resulting from LTB(4) challenge of the skin, again showing a dose-response relationship. In view of the large variation in the elimination half-life of VML 295 (25-88.5 h) in individual human subjects, the present pharmacological study was designed to provide information on the pharmacodynamics of the drug by the assessment of VML 295 plasma concentrations, ex vivo LTB(4)-induced CD11b upregulation of neutrophils, neutrophil accumulation in the skin following epicutaneous application of LTB(4) and epidermal regeneration following standardized surface trauma. A group of 36 healthy volunteers were treated in a double-blind study with VML 295 at 200 mg twice daily, VML 295 at 200 mg once daily or placebo for 7 days. Before treatment, at the end of treatment and following discontinuation of treatment, VML 295 plasma concentrations and CD11b upregulation of blood neutrophils were assessed. In 18 subjects, the effects of the three treatments on LTB(4)-induced inflammatory were assessed before and at the end of treatment, and in the remaining 18 subjects the effects of these treatments on epidermal regeneration were assessed similarly. VML 295 at 200 mg either twice or once daily has a profound inhibitory effect on ex vivo LTB(4)-induced CD11b upregulation of blood neutrophils, LTB(4)-induced neutrophil accumulation in the skin, trauma-induced hyperproliferation of the epidermis and regenerative keratinization. The twice daily dose schedule was significantly more effective than the once daily regimen in reducing ex vivo CD11b stimulation of neutrophils, in blood samples collected 24 h after discontinuation of VML 295 treatment. The twice daily schedule tended to be more efficient in skin biopsies, although this difference was not statistically significant in the number of subjects investigated. A plasma concentration of 100 ng/ml proved to be the threshold for these effects. The profound biological effects, both systemically and cutaneously, as well as the safety profile, make VML 295 a promising drug for skin disorders characterized by epidermal proliferation and neutrophil accumulation.

Investigation on a novel and specific leukotriene B4 receptor antagonist in the treatment of stable plaque psoriasis.

The aim of the present study was to investigate the efficacy and clinical tolerability of the specific leukotriene B4 receptor antagonist VML295 in the treatment of stable plaque psoriasis. Immunohistochemical and flow cytometrical methods were used to assess the effects on inflammation and epidermal proliferation. VML295 in the treatment of chronic plaque psoriasis was shown to be safe and well tolerated. After treatment, there was a statistically significant difference between patients treated with VML295 and patients treated with placebo with respect to the leukotriene B4-induced CD11b up-regulation on the cell surface of polymorphonuclear leukocytes derived from peripheral blood. Ex vivo CD11b up-regulation in the VML295-treated group was completely inhibited after 7 days of treatment (P = 0.001). This effect persisted until the end of the treatment period (P = 0.004 on day 15 and P < 0.0001 after 4 weeks), whereas CD11b up-regulation in the placebo group remained unaffected. There was no statistically significant difference in the median psoriasis area and severity index between the treatment groups at the end of the treatment period. During treatment, no significant histological changes were observed in the markers for cutaneous inflammation and epidermal proliferation. Although not statistically significant, a tendency for the increased expression of some markers of cutaneous inflammation and epidermal proliferation was observed after 1 week of treatment with VML295, and a decreased expression of these markers was seen after 4 weeks of treatment with VML295. This observation could indicate anti-inflammatory effects of VML295 appearing between 2 and 4 weeks after the start of treatment.

The efficacy of a new topical treatment for psoriasis: Mirak.

BACKGROUND: A new natural product for the treatment of psoriasis has recently become available in many European countries: Mirak. The Mirak Home Care Packs consist of natural spring water, volcanic earth and vitamin E cream. Recently, the efficacy of Mirak has come into question. As this treatment is used by many psoriasis patients in Europe, it is important for dermatologists to be informed about the clinical effects of the therapy. AIM: To evaluate the efficacy and side effects of the Mirak Home Care Packs. METHODS: By means of a placebo-controlled left/right comparison, both clinical and histological parameters were evaluated during 6 weeks of treatment. RESULTS: The reduction in induration was significantly greater in the Mirak-treated lesions than in the lesions treated with a placebo. A reduction in desquamation was found in both treatments; the difference between the treatments was not statistically significant. A decrease in number of proliferative cells in the Mirak-treated lesions was seen, but the difference with placebo-treated lesions was not significant. The other investigated parameters did not change during treatment. No side effects were seen. CONCLUSIONS: The Mirak Home Care Pack induces a modest therapeutic effect compared to placebo treatment, without significant side effects. Treatment with the Mirak Home Care Packs alone will probably not be able to compete with the already existing treatments for psoriasis.

Effects of systemic treatment with liarozole on cutaneous inflammation, epidermal proliferation and differentiation in extensive plaque psoriasis.

Liarozole is a novel inhibitor of the enzyme cytochrome P450 which has inhibitory effects on the 4-hydroxylation of retinoic acid. Previous studies have shown that liarozole is effective in the treatment of psoriasis. We performed an immunohistochemical study on the lesional skin from 7 patients with extensive plaque psoriasis, who were treated with systemic liarozole 75 mg BID for a period of at least 2 months. The effects of liarozole treatment on clinical and histological parameters were investigated. In particular, the effect of liarozole on the integrin markers CD11b and CD18 was studied. For immunohistochemistry, three consecutive biopsies were taken: before treatment, after 4, and after 8 weeks of treatment. Clinical scores and side effects were recorded before and during treatment. The medication was well tolerated and only mild side effects were reported, which were comparable with hypervitaminosis A. After 2 months of treatment a statistically significant decrease of the extent of body involvement was observed. In the psoriatic plaque, markers for epidermal proliferation and cutaneous inflammation decreased, and markers for epidermal differentiation increased to values comparable to normal skin. The first therapeutic effects in the psoriatic plaque occurred after 4 weeks of treatment, and consisted of a decreased induration, accompanied by a decrease of the total number of inflammatory infiltrate cells and a decreased epidermal ICAM-1 expression. Already after 4 weeks of treatment, a decrease of CD11b-positive cells was observed. Subsequently, after 8 weeks of treatment recruitment of cycling epidermal cells and the number of involucrin-positive cell layers decreased. The present study demonstrates that liarozole treatment of psoriasis results in a reduction of aspects of cutaneous inflammation and subsequently a reduction of epidermal proliferation and promotion of differentiation. After 4 weeks of treatment, effects are observed on the epidermal ICAM-1 expression and on the CD11b-positive cell population.

Transient and stable expression of gusA fusions with rice genes in rice, barley and perennial ryegrass.

Transcriptional and translational fusions were made between the reading frame coding for beta-D-glucuronidase and sequences of either a constitutively expressed rice gene (GOS2) involved in initiation of translation or a light-inducible rice gene (GOS5). The transient expression of the fusions was studied via particle bombardment of seedling tissues of rice, perennial ryegrass and barley. Furthermore, the results of transient and stable expression were compared for cell suspensions of four rice varieties, one barley variety and one perennial ryegrass variety. The GOS2-gusA fusions were active in all three monocots studied. Best results were obtained for a construct having both a transcriptional and a translational fusion as well as intron and exon sequences (PORCEHyg). The level of GUS activity was in the range of activities as obtained by the 35S CaMV promoter transcriptionally fused to gusA. The gusA fusion with the light-inducible gene (GOS5) was active in green seedling tissues of all monocots studied. Also a weak expression compared to the GOS2 constructs was found in stably transformed rice callus. The gusA fusions with the mannopine synthase promoters 1' and 2' of the TR-DNA were transiently expressed at lower levels in cell suspensions than PORCEHyg. For stably transformed rice callus the expression of the GOS2-gusA fusion often decreased during prolonged subculture. This decrease in GUS activity and the various GUS-staining phenotypes of transgenic calli are explained by the presence of different cell types in the suspensions used and in the calli. It is presumed that the nature of the cells and their relative contribution in the calli change drastically upon further subculture.

Transient and stable expression of gusA fusions with rice genes in rice, barley and perennial ryegrass.

Transcriptional and translational fusions were made between the reading frame coding for beta-D-glucuronidase and sequences of either a constitutively expressed rice gene (GOS2) involved in initiation of translation or a light-inducible rice gene (GOS5). The transient expression of the fusions was studied via particle bombardment of seedling tissues of rice, perennial ryegrass and barley. Furthermore, the results of transient and stable expression were compared for cell suspensions of four rice varieties, one barley variety and one perennial ryegrass variety. The GOS2-gusA fusions were active in all three monocots studied. Best results were obtained for a construct having both a transcriptional and a translational fusion as well as intron and exon sequences (PORCEHyg). The level of GUS activity was in the range of activities as obtained by the 35S CaMV promoter transcriptionally fused to gusA. The gusA fusion with the light-inducible gene (GOS5) was active in green seedling tissues of all monocots studied. Also a weak expression compared to the GOS2 constructs was found in stably transformed rice callus. The gusA fusions with the mannopine synthase promoters 1' and 2' of the TR-DNA were transiently expressed at lower levels in cell suspensions than PORCEHyg. For stably transformed rice callus the expression of the GOS2-gusA fusion often decreased during prolonged subculture. This decrease in GUS activity and the various GUS-staining phenotypes of transgenic calli are explained by the presence of different cell types in the suspensions used and in the calli. It is presumed that the nature of the cells and their relative contribution in the calli change drastically upon further subculture.

Early changes in the glycopeptides of human B-lymphocytes after Epstein-Barr virus infection in vitro.

Human B lymphocytes obtained from healthy donors were infected with Epstein-Barr virus in vitro. From the initiation of infection to the final establishment of a permanent lymphoblastoid cell line, fucosyl glycopeptides of the cell surface were investigated. In order of appearance the following events took place: expression of Epstein-Barr virus-determined nuclear antigen, mitotic activity of the cells and specific glycopeptide alterations on the cell surface. This specific alteration in glycopeptides, as determined by gel filtration, is manifested by the appearance of fast-eluting glycopeptides and was similar to that found on Burkitt lymphoma cells. Neither pokeweed-mitogen-stimulated B lymphocytes nor exponentially growing normal T lymphocytes exposed fast-eluting glycopeptides on their surfaces. Therefore, it is concluded that the appearance of these fast-eluting glycopeptides on the surface of lymphoblastoid cells after EBV infection is not the result of culture conditions or conditions of growth as such. The similarity with glycopeptides derived from Burkitt lymphoma cells, and the observation that a considerable proportion of the B cells becomes immortalized, are discussed.