Systematic review and meta-analysis of preoperative imaging prediction of renal cell carcinoma tumour thrombus inferior vena cava wall invasion.

AIM: To summarise published evidence assessing the preoperative diagnostic performance of identifying inferior vena cava (IVC) wall invasion in patients with renal cell carcinoma (RCC). MATERIALS AND METHODS: A systematic approach was used to identify studies that assessed IVC wall invasion with non-invasive imaging preoperatively. Search limits included English language and human study participants. A meta-analysis was conducted using random effects models to compare radiographic vascular size parameters and the association of IVC wall invasion. RESULTS: A total of 15 studies were identified, which included computed tomography (CT), magnetic resonance imaging (MRI), positron-emission tomography (PET), and ultrasound assessment. In the majority of cases, CT and MRI was utilised with high accuracy in predicting IVC wall invasion. A meta-analysis of commonly reported radiographic vascular size parameters found that IVC wall invasion was associated with greater IVC maximum anteroposterior (AP) diameter (mean difference [MD] = 6.58 mm, 95% confidence interval [CI]: 2.84-10.33, p=0.0006) and IVC maximum AP diameter at the level of the renal vein ostium (MD = 5.69 mm, 95% CI: 4.35-7.03, p<0.0001). Renal vein maximum AP dimension was not associated with IVC wall invasion (MD = 2.56 mm, 95% CI: -0.46-5.58, p=0.10). CONCLUSION: Multi-technique work-up, specifically CT and MRI and reporting of vascular radiographic parameters, of RCC patients with IVC tumour thrombus may be useful in predicting IVC wall invasion, thereby allowing appropriate surgical planning and patient education.

Outcomes in prevention and management of miscarriage trials: a systematic review.

BACKGROUND: There is a substantial body of research evaluating ways to prevent and manage miscarriage, but all studies do not report on the same outcomes. OBJECTIVE: To review systematically, outcomes reported in existing miscarriage trials. SEARCH STRATEGY: MEDLINE, Embase, CINAHL, and Cochrane were searched from inception until January 2017. SELECTION CRITERIA: Randomised controlled trials (RCTs) reporting prevention or management of miscarriage. Miscarriage was defined as a pregnancy loss in the first trimester. DATA COLLECTION AND ANALYSIS: Data about the study characteristics, primary, and secondary outcomes were extracted. MAIN RESULTS: We retrieved 1553 titles and abstracts, from which 208 RCTs were included. For prevention of miscarriage, the most commonly reported primary outcome was live birth and the top four reported outcomes were pregnancy loss/stillbirth (n = 112), gestation of birth (n = 68), birth dimensions (n = 65), and live birth (n = 49). For these four outcomes, 58 specific measures were used for evaluation. For management of miscarriage, the most commonly reported primary outcome was efficacy of treatment. The top four reported outcomes were bleeding (n = 186), efficacy of miscarriage treatment (n = 105), infection (n = 97), and quality of life (n = 90). For these outcomes, 130 specific measures were used for evaluation. CONCLUSIONS: Our review found considerable variation in the reporting of primary and secondary outcomes along with the measures used to assess them. There is a need for standardised patient-centred clinical outcomes through the development of a core outcome set; the work from this systematic review will form the foundation of the core outcome set for miscarriage. TWEETABLE ABSTRACT: There is disparity in the reporting of outcomes and the measures used to assess them in miscarriage trials.

What are the validated animal models for tendinopathy?

Chronic tendinopathy refers to a broad spectrum of pathological conditions in tendons and their insertion, with symptoms including activity-related chronic pain. To study the pathogenesis and management strategies of chronic tendinopathy, studies in animal models are essential. The different animal models in the literature present advantages and limitations, and there is no consensus regarding the criteria of a universal tendinopathy animal model. Based on the review of literature and the discussion in the International Symposium on Ligaments and Tendons-X, we concluded that established clinical, histopathological and functional characteristics of human tendinopathy were all important and relevant criteria to be met, if possible, by animal models. As tendinopathy is a progressive, multifactorial tendon disorder affecting different anatomical structures, it may not be realistic to expect a single animal model to study all aspects of tendinopathy. Staging of tendinopathy over time and clearer definition of tendinopathies in relation to severity and type would enable realistic targets with any animal model. The existing animal models can be used for answering specific questions (horses for courses) but should not be used to conclude the general aspects of tendinopathy neither in animals nor in human.

Optimisation of bone marrow aspiration from the equine sternum for the safe recovery of mesenchymal stem cells.

REASONS FOR PERFORMING STUDY: Mesenchymal stem cell (MSC) therapy for orthopaedic disease is being used with increasing frequency; there is a need to define a safe, reliable and effective technique for the recovery of MSCs from the sternum of the horse. OBJECTIVES: To describe an optimised safe technique for obtaining bone marrow-derived MSCs from the sternum of the Thoroughbred horse. METHODS: The anatomical relationship of the sternum with the heart and internal anatomy was demonstrated in cadavers. Sternal anatomy was evaluated ultrasonographically and after midline sectioning. Sternebrae were examined histologically after aspiration to determine the effect of needle insertion. The quality of the aspirate was evaluated as the number of colony-forming units from sequential and separately aspirated 5 ml aliquots and assessed for their multipotency using trilineage differentiation. RESULTS: The optimal safe location for the needle was the 5th sternebra because it had a safe dorsoventral thickness and was cranial to the apex of the heart. This sternebra could be reliably identified ultrasonographically. Aspirates could also be obtained from the 4th and 6th sternebrae, although the former is between the front limbs and the latter closer to the heart. Minimal disruption of the internal bony architecture was seen after needle insertion through the thin outer cortex and the first 5 ml aliquot contained the greatest number of colony-forming units of mesenchymal stem cells with trilineage capabilities. CONCLUSIONS: Accurate placement of a Jamshidi needle into the medullary cavity of the 4th-6th individual sternebrae is facilitated by the use of ultrasonography and enables aspiration of bone marrow reliably with minimal damage to the sternum and risk to the horse. POTENTIAL CLINICAL RELEVANCE: Sternal marrow aspiration as described is a safe and reliable technique to obtain MSCs for orthopaedic cell-based therapies.

Cell-based therapies for tendon and ligament injuries.

Tendon and ligament injuries have proved difficult to treat effectively. Cell-based therapies offer the potential to harness the complex protein synthetic machinery of the cell to induce a regenerative response rather than fibrous scarring. This article reviews the current state of play with respect to the clinically used cell preparations for the treatment of tendon and ligaments overstrain injuries.

Evaluation of a filter-prepared platelet concentrate for the treatment of suspensory branch injuries in horses.

OBJECTIVES: Platelet preparations have become a treatment for soft tissue injuries in horses. This study evaluated a novel filter-based system to concentrate platelets and assesses its value in the treatment of suspensory ligament branch desmitis. METHODS: Filtered platelet concentrate was prepared from 55 ml of venous blood obtained from 21 normal horses. Platelets and white blood cells in whole blood and filtered platelet concentrate were measured, as was platelet activating factor (PAF)-induced platelet-derived growth factor-BB (PDGF-BB) release. Eleven horses with 18 focal acute suspensory ligament branch injuries were treated intra-lesionally with autologous filtered platelet concentrate and evaluated clinically and ultrasonographically for one to three years. RESULTS: The increase in concentration of platelets in the filtered concentrate in comparison with whole blood (6.9 ± 1.9-fold) was significantly greater than the increase in white blood cells (3.8 ± 0.8-fold) (p <0.0001). There was no effect of sex or breed on platelet concentration. Platelets were responsive to PAF with >100-fold higher levels of PDGF release over basal levels. All hypoechoic lesions re-evaluated within three months had resolved. Five of the 11 horses returned to their previous level of work, one was exercising at a lower level, three were retired, one died for unrelated reasons, and one was still convalescing. CLINICAL SIGNIFICANCE: Filtered platelet concentrate was easily and reliably prepared and injected into suspensory ligament branch injuries without short-term complications. This treatment was associated with rapid resolution of ultrasonographic lesions and lameness. Filtered platelet concentrate represents a convenient alternative for the treatment of suspensory ligament branch injuries.

Clinical update on the use of mesenchymal stem cells in equine orthopaedics.

Stem cells have received much attention in recent times because of their potential to improve healing of othropaedic problems. This manuscript presents the genesis, issues and current state of stem cell treatment in equine medicine. Current literature supports the use of mesenchymal stem cells (MSCs) for treatment of orthopaedic problems.

Tiludronate infusion in the treatment of bone spavin: a double blind placebo-controlled trial.

REASONS FOR PERFORMING STUDY: Tiludronate regulates bone remodelling through a decrease of the resorptive process and should therefore ameliorate the remodelling processes active in osteoarthritis of the distal tarsal joints ('bone spavin') and alleviate pain associated with abnormal bone lysis. OBJECTIVE: To confirm the efficacy of tiludronate, administered as a single infusion at a dose of 1 mg/kg bwt, in the treatment of bone spavin in the horse. METHODS: A double blind placebo controlled trial on 108 clinical cases of bone spavin was undertaken. The lameness score of the lamest limb was assessed following distal tarsal analgesia of the contralateral limb and followed-up using the same procedure throughout the study. Bone spavin in the lamest limb was confirmed by distal tarsal analgesia and radiography. Horses were treated at Day 0 and reassessed 60 days later after controlled exercise. A second nonblinded treatment was given to unresponsive horses and all horses were re-examined at Day 120. Exercise levels were recorded at each examination. RESULTS: Eighty-seven horses completed the trial as per the protocol. The tiludronate horses were significantly less lame than the placebo horses (P = 0.0318). Horses treated at Day 60 with tiludronate showed further improvement in lameness at Day 120 (P = 0.0096 and P = 0.0034 for horses treated with tiludronate and placebo at Day 0, respectively). The only significant difference in radiographic findings between tiludronate and placebo was for presence of periarticular osteophytes (P = 0.006). CONCLUSIONS: Tiludronate treatment is proven to be effective in bone spavin in horses in association with a controlled exercise programme. CLINICAL RELEVANCE: Tiludronate in combination with controlled exercise offers an alternate medical treatment for bone spavin.

TPO antibody positivity and adverse pregnancy outcomes.

Thyroid autoimmunity (TAI) is prevalent amongst women of reproductive age. TAI describes the presence of circulating anti-thyroid autoantibodies that are targeted against the thyroid, with or without thyroid dysfunction. Thyroid peroxidase antibodies (TPOAb) are the most common anti-thyroid autoantibodies. Around 10% of biochemically euthyroid individuals also have an elevated TPOAb titre. Many studies have linked the presence of TPOAb to adverse maternal and fetal outcomes in pregnancy, in particular miscarriage and pre-term birth, even in the absence of thyroid dysfunction. The causal pathway is poorly understood and few trials have looked to find treatments to reduce adverse outcomes. This review discusses in detail the associated adverse outcomes of TPOAb in pregnancy and the results of trials exploring methods to reduce such outcomes. Recommendations for counselling and monitoring of women with TPOAb and suggested areas for future work are also outlined.

Improved diagnostic criteria for digital flexor tendon sheath pathology using contrast tenography.

BACKGROUND: Pathology of the digital flexor tendon sheath is a significant cause of lameness in the horse. Imaging is important to identify lesions and inform on prognosis prior to tenoscopic surgery. OBJECTIVES: To use a large population to evaluate 1) the sensitivity and specificity of digital flexor tendon sheath (DFTS) contrast radiographs in diagnosing manica flexoria (MF) tears, deep digital flexor tendon (DDFT) tears and constriction of the palmar/plantar annular ligament (PAL) using novel criteria; 2) predisposition to pathology in signalment and limb affected. STUDY DESIGN: Multicentre retrospective cohort study. METHODS: The medical records of 206 horses with lameness localised to the DFTS, contrast radiographs and subsequent tenoscopic surgery were reviewed. Breed and limb predispositions were evaluated for pathology of the DDFT, MF and PAL constriction. Contrast radiographs of the DFTS were reviewed by four masked operators and for each pathology the sensitivity, specificity and interobserver variability were calculated. RESULTS: Contrast tenography was a sensitive test for MF tears (92% confidence interval [CI] 88.4-94.4%; specificity 56%, CI 51.1-61.1%) and specific for diagnosing DDFT tears (73%, CI 68.6-76.8%; sensitivity 54%, CI 47.8-60.2%) but had a lower sensitivity (71%, CI 65.1-75.9% ) and specificity (45%, CI 39.1-52.0%) for PAL constriction. It had good to substantial interobserver agreement for MF and DDFT tears (Krippendorff's alpha 0.68 and 0.46 respectively). Ponies (57%) and cobs (58%) were significantly more likely to be affected with MF tears (other breeds 20-39%, P = 0.003) and Thoroughbreds (50%), warmbloods (45%) and draught breeds (48%) were more likely to have DDFT tears (other breeds 22-34%, P = 0.01). MF tears and PAL constriction were overrepresented in the hindlimbs compared to DDFT tears in forelimbs. MAIN LIMITATIONS: No standardisation of contrast radiographs was possible. The subjectivity of diagnosis of PAL constriction may also have led to bias. Radiographs were read as JPEGS reducing ability to manipulate images. CONCLUSIONS: Contrast radiography of the DFTS is accurate in the pre-operative diagnosis of DFTS pathologies. Different pathologies are overrepresented in certain breeds and limbs.

A novel role for the integrin-binding III-10 module in fibronectin matrix assembly.

Fibronectin matrix assembly is a cell-dependent process which is upregulated in tissues at various times during development and wound repair to support the functions of cell adhesion, migration, and differentiation. Previous studies have demonstrated that the alpha 5 beta 1 integrin and fibronectin's amino terminus and III-1 module are important in fibronectin polymerization. We have recently shown that fibronectin's III-1 module contains a conformationally sensitive binding site for fibronectin's amino terminus (Hocking, D.C., J. Sottile, and P.J. McKeown-Longo. 1994. J. Biol. Chem. 269: 19183-19191). The present study was undertaken to define the relationship between the alpha 5 beta 1 integrin and fibronectin polymerization. Solid phase binding assays using recombinant III-10 and III-1 modules of human plasma fibronectin indicated that the III-10 module contains a conformation-dependent binding site for the III-1 module of fibronectin. Unfolded III-10 could support the formation of a ternary complex containing both III-1 and the amino-terminal 70-kD fragment, suggesting that the III-1 module can support the simultaneous binding of III-10 and 70 kD. Both unfolded III-10 and unfolded III-1 could support fibronectin binding, but only III-10 could promote the formation of disulfide-bonded multimers of fibronectin in the absence of cells. III-10-dependent multimer formation was inhibited by both the anti-III-1 monoclonal antibody, 9D2, and amino-terminal fragments of fibronectin. A fragment of III-10, termed III-10/A, was able to block matrix assembly in fibroblast monolayers. Similar results were obtained using the III-10A/RGE fragment, in which the RGD site had been mutated to RGE, indicating that III-I0/A was blocking matrix assembly by a mechanism distinct from disruption of integrin binding. Texas red-conjugated recombinant III-1,2 localized to beta 1-containing sites of focal adhesions on cells plated on fibronectin or the III-9,10 modules of fibronectin. Monoclonal antibodies against the III-1 or the III-9,10 modules of fibronectin blocked binding of III-1,2 to cells without disrupting focal adhesions. These data suggest that a role of the alpha 5 beta 1 integrin in matrix assembly is to regulate a series of sequential self-interactions which result in the polymerization of fibronectin.

Variants within the MMP3 gene are associated with Achilles tendinopathy: possible interaction with the COL5A1 gene.

OBJECTIVES: Sequence variation within the COL5A1 and TNC genes are known to associate with Achilles tendinopathy. The primary aim of this case-control genetic association study was to investigate whether variants within the matrix metalloproteinase 3 (MMP3) gene also contributed to both Achilles tendinopathy and Achilles tendon rupture in a Caucasian population. A secondary aim was to establish whether variants within the MMP3 gene interacted with the COL5A1 rs12722 variant to raise risk of these pathologies. METHODS: 114 subjects with symptoms of Achilles tendon pathology and 98 healthy controls were genotyped for MMP3 variants rs679620, rs591058 and rs650108. RESULTS: As single markers, significant associations were found between the GG genotype of rs679620 (OR = 2.5, 95% CI 1.2 to 4.90, p = 0.010), the CC genotype of rs591058 (OR = 2.3, 95% CI 1.1 to 4.50, p = 0.023) and the AA genotype of rs650108 (OR = 4.9, 95% CI 1.0 to 24.1, p = 0.043) and risk of Achilles tendinopathy. The ATG haplotype (rs679620, rs591058, and rs650108) was under-represented in the tendinopathy group when compared to the control group (41% vs 53%, p = 0.038). Finally, the G allele of rs679620 and the T allele of COL5A1 rs12722 significantly interacted to raise risk of AT (p = 0.006). No associations were found between any of the MMP3 markers and Achilles tendon rupture. CONCLUSION: Variants within the MMP3 gene are associated with Achilles tendinopathy. Furthermore, the MMP3 gene variant rs679620 and the COL5A1 marker rs12722 interact to modify the risk of tendinopathy. These data further support a genetic contribution to a common sports related injury.

Ultrasonographic assessment of the superficial digital flexor tendons of National Hunt racehorses in training over two racing seasons.

REASONS FOR PERFORMING STUDY: It is important to ascertain the prevalence of superficial digital flexor tendon (SDFT) injuries and to improve methods of predicting injury in National Hunt (NH) racehorses. OBJECTIVES: To establish: 1) the prevalence of SDFT tendinopathy in NH horses; 2) whether routine ultrasonography can be used to predict SDFT injuries; 3) whether previous tendinopathy predisposes to reinjury; 4) a normal range for the SDFT cross-sectional area (CSA); and 5) the effects of gender, age, background (ex-flat or ex-store), limb, training and rest periods on SDFT CSA. METHODS: Routine ultrasound assessment of the palmar metacarpal soft tissues of 263 NH racehorses was performed on up to 6 occasions over 2 NH racing seasons. RESULTS: The prevalence of SDFT pathology detected using ultrasonography was 24% (n = 148), with a nonsignificant variation between yards of 10-40%. No changes in SDFT CSA or ultrasonographic appearance were detected prior to injury. Older horses had a significantly higher prevalence of SDFT pathology compared to younger horses, and horses with tendinopathy were more likely to suffer an acute injury compared to horses with no evidence of pathology. A reference range for normal CSA measurements was established as 77-139 mm2 at level 4, from 142 horses with no ultrasonographic evidence of SDFT pathology. The CSA of normal horses did not vary significantly with age, limb or over 2 racing seasons, but did with sex and background. CONCLUSIONS: The study confirms that SDFT tendinopathy is common in NH horses, with substantial variation between training yards. Ultrasonography at 3 month intervals did not seem to predict acute SDFT injuries. POTENTIAL RELEVANCE: Variation in the prevalence of tendinopathy between yards suggests that training methods may influence injury rate. It was not possible to predict injury using routine ultrasonography and therefore other methods must be identified. A normal reference range for SDFT CSA is provided.

Descriptive epidemiology of fracture, tendon and suspensory ligament injuries in National Hunt racehorses in training.

REASONS FOR PERFORMING STUDY: While fractures and tendon injuries are known to be important diseases in National Hunt (NH) racehorses during racing, there are no accurate estimates of their incidence in NH training yards. OBJECTIVES: To estimate the incidence of fractures and tendon and suspensory ligament injuries (TLIs) in NH racehorses in training; to describe the injuries incurred and to compare injury incidence rates by horse age, trainer, gender and background (ex-flat vs. ex-store horses). METHODS: Cohort data were collected from 14 UK NH training yards for 2 racing seasons. Daily exercise regimens and details of fractures and TLIs occurring in study horses were recorded. RESULTS: Data were gathered from 1223 horses that spent 9466 months at risk of injury. The fracture incidence rate was 1.1/100 horse months and varied significantly by trainer (P<0.001) but not by gender, age or background. The pelvis and third metacarpal bone (MCIII) were the most common fracture sites, although this varied between racing and training. The TLI incidence rate was 1.9/100 horse months and varied significantly by trainer (P = 0.05) and age (P<0.001) but not by gender or background. However, ex-store horses were significantly more likely to have a TLI on the racecourse than ex-flat horses (P = 0.01). Superficial digital flexor injuries accounted for 89% of all TLIs, the remainder being suspensory ligament injuries. CONCLUSIONS AND POTENTIAL RELEVANCE: Fractures and TLIs are important causes of morbidity and mortality in NH racehorses in training in England. This study provides accurate estimates of their incidence in this population and provides a baseline against which to monitor the effect of future interventions.

Can exercise modulate the maturation of functionally different immature tendons in the horse?

Tendons can be considered in two functional groups, those contributing to energetics of locomotion and those acting solely to position the limb. The energy-storing tendons in both human and equine athletes have a high frequency of injury with similar pathophysiology. In previous studies, high-intensity exercise appears to induce a disruption of the matrix rather than functional adaptation in adults. Here we explore the hypothesis that the introduction of controlled exercise during growth would result in an adaptive response without deleterious effects. Young horses were given a controlled exercise program similar to that previously shown to induce matrix changes in energy-storing tendons of skeletally mature animals. The tendons were assessed in relation to mechanical properties, molecular composition, and morphology. Results showed a significant increase in cartilage oligomeric matrix protein (COMP) in the positional tendon but not in the energy-storing tendon. Other matrix properties and mechanical properties were not significantly changed. While the imposition of high-strain-rate exercise in immature horses failed to augment the development of the energy-storing tendon over and above that induced by normal pasture exercise, it did not induce deleterious changes, supporting an earlier introduction of athletic training in horses.

The influence of exercise during growth on ultrasonographic parameters of the superficial digital flexor tendon of young Thoroughbred horses.

REASON FOR PERFORMING STUDY: Conditioning by early training may influence the composition of certain musculoskeletal tissues, but very few data exist on its effect during growth on tendon structure and function. OBJECTIVES: To investigate whether conditioning exercise in young foals would lead to any ultrasonographically detectable damage to the superficial digital flexor tendon or an increase in cross-sectional area (CSA). METHODS: Thirty-three Thoroughbred foals reared at pasture were allocated to 2 groups: control (PASTEX) allowed exercise freely at pasture; and CONDEX, also at pasture, began conditioning exercise from mean age 21 days over 1030 m on a purpose-built oval grass track, for 5 days/week until mean age 18 months. Foals were observed daily, and underwent orthopaedic examination monthly. Ultrasonographic images of the superficial digital flexor tendon (SDFT) at the mid-metacarpal level of both forelimbs were obtained in all foals at ages 5, 8, 12, 15 and 18 months. CSA was validated (r(2) = 0.89) by determining CSA from digital photographs of the transected SDFT surface from 12 of the horses necropsied at age 17.1 months. RESULTS: here was no clinical or ultrasonographic evidence of tendonopathy in either group and the greatest increase in mean CSA in both groups occurred between age 5 and 8 months. Across all age categories, there was no significant difference in mean CSA between the left and right limbs, or colts and fillies; there was a trend towards a larger CSA in the CONDEX group (P = 0.058). CONCLUSIONS: There was no conclusive evidence for a structural adaptive hypertrophy of the SDFT, probably because the regimen was insufficiently rigorous or because spontaneous pasture exercise may induce maximal development of energy storing tendons. POTENTIAL RELEVANCE: A moderate amount of early conditioning exercise against a background of constant exercise at pasture is not harmful to the development of the flexor tendons.

Evaluation of a new strategy to modulate skeletal development in racehorses by imposing track-based exercise during growth: the effects on 2- and 3-year-old racing careers.

REASON FOR PERFORMING STUDY: No data exist on the effects of conditioning exercise at foal age on workload and subsequent clinical injury rate during their 2- and 3-year-old racing careers. OBJECTIVES: To investigate the effects of subjecting TB foals to conditioning exercise prior to the start of race training on: the workload required to reach a level of fitness sufficient to compete; and the prevalence of orthopaedic injury during the first 2 seasons of their racing careers. METHODS: Twenty 18-month-old TBs, 12 subjected to conditioning exercise at foal age (CONDEX) and 8 exercised spontaneously at pasture only (PASTEX) were trained and entered in competitive events. Workload was quantified using the cumulative workload index (the product of average velocity and distance at a specific gait) and the animals were monitored clinically and radiographically for signs of musculoskeletal disorders. RESULTS: Workload to reach the desired fitness level was similar for CONDEX and PASTEX. CONDEX performed more prerace training sessions as 2-year-olds (P<0.05). The incidence of orthopaedic injuries was low in both groups and there were no differences in the occurrence of orthopaedic ailments. PASTEX animals tended to show signs of musculoskeletal disorders earlier than CONDEX animals. This time difference was significant for metacarpophalangeal joint pain on flexion, reduced carpal flexion and hindlimb lameness (P<0.05). CONCLUSIONS: Subjecting TB foals to conditioning exercise early in life does not have adverse effects on racing careers at ages 2 and 3 years, and does not influence the workload needed to reach a fitness level that is sufficient for racing. POTENTIAL RELEVANCE: The lack of negative effects and the indications of some positive effects of early conditioning exercise in the Thoroughbred encourage further large-scale comparative studies.

Evaluation of a new strategy to modulate skeletal development in Thoroughbred performance horses by imposing track-based exercise during growth.

REASON FOR PERFORMING STUDY: No data exist on the intensity of exercise required or on possible harmful effects of increasing exercise in foals over the natural level when free at pasture. OBJECTIVES: To investigate whether an increase in workload over free pasture exercise in the period from directly after birth to the start of training is tolerated by Thoroughbred (TB) foals without increasing injury rate or producing other undesired side effects. METHODS: Thirty-three TB foals were allocated to one of 2 exercise groups directly after birth. One group (PASTEX) was raised on pasture and the other (CONDEX) kept under identical circumstances, but was additionally subjected to an exercise protocol of gradually increasing intensity. Foals were monitored periodically and scored for the presence of clinical signs related to the musculoskeletal system (joint effusion, pain at flexion, occurrence of physeal swelling), and radiographs taken at the end of the conditioning phase. Also, behavioural studies were performed to detect any changes in behaviour related to the exercise programme. Cortisol levels were measured in both groups, to assess the level of stress. RESULTS: Workload in the CONDEX group was significantly higher than in the PASTEX group (approximately 30%). Conditioning increased the likelihood for joint effusion in the antebrachiocarpal joint, but reduced tarsocrural effusion and physeal swelling at the lateral distal radius, the third metacarpal bone (medial aspect) and lateral and medial aspects of the third metatarsal bone. CONCLUSIONS: The 30% increase in workload did not affect the animals' welfare, effects of conditioning exercise on clinical musculoskeletal health were few and there were no adverse effects. POTENTIAL RELEVANCE: This study supports the feasibility of imposing early conditioning exercise in horses and is a benchmark for its effects on the development of equine musculoskeletal tissues.

Transcriptional down-regulation of N-myc expression during B-cell development.

We have used nuclear run-on and DNase I sensitivity analyses to study the activity of the N-myc genes in cell lines that represent different stages of B-cell development. Both transformed pre-B-cell lines and a nontransformed pre-B-cell clone transcribe the N- and c-myc genes at substantial levels; in the nontransformed clone, transcription of these genes is regulated by the pre-B-cell growth factor interleukin-7. In contrast, transformed cell lines that represent the more mature stages of the B-cell pathway and mitogen-stimulated normal splenic B lymphocytes express the c-myc gene but do not express the N-myc gene at detectable levels. Down-regulation of N-myc expression in these cells occurs at the level of transcriptional initiation. Correspondingly, a set of DNase I-hypersensitive sites present in the 5' region of the N-myc promoter of pre-B-cell lines are absent in B-cell lines. To further elucidate this process, we have constructed fusion cell lines between an N-myc-expressing pre-B-cell line and a nonexpressing myeloma line; the hybrid cell lines transcriptionally down-regulate the pre-B copies of the N-myc gene. Lack of N-myc expression in a number of nonlymphoid cell lines also resulted from lack of N-myc transcription. Together, our findings demonstrate that the down-regulation of N-myc expression in the later stages of B-cell development is mediated primarily at the level of transcriptional initiation. They further show that dominant, trans-acting factors present in more mature B-lineage cell lines act to down-regulate the transcription of N-myc.

Mechanism of endogenous myc gene down-regulation in E mu-N-myc tumors.

Transgenic mouse lines carrying the N-myc oncogene deregulated by the immunoglobulin heavy-chain enhancer spontaneously develop B-lymphoid tumors (R. Dildrop, A. Ma, K. Zimmerman, E. Hsu, A. Tesfaye, R. DePinho, and F. W. Alt, EMBO J. 8:1121-1128, 1989; H. Rosenbaum, E. Webb, J. M. Adams, S. Cory, and A. W. Harris, EMBO J. 8:749-755). Permanent cell lines derived from these tumors (E mu-N-myc cell lines) express extremely high levels of the N-myc transgene but little or no detectable endogenous N-myc or c-myc. We have employed nuclear run-on assays to show that down-regulation of endogenous N- and c-myc expression occurs at the transcriptional level. To determine whether the lack of endogenous myc gene transcription is a direct effect of high-level N-myc transgene expression, we have generated Abelson murine leukemia virus (A-MuLV)-transformed cell lines from prelymphomatous E mu-N-myc mice (A-MuLV/E mu-N-myc cell lines). Although these A-MuLV/E mu-N-myc lines express very high levels of the N-myc transgene, they continue to transcribe the endogenous c-myc gene. These findings demonstrate that high-level N-myc gene expression alone does not necessarily lead to down-regulation of endogenous myc gene expression and suggest that events associated with transformation by N-myc may be critical to this process.