Osteochondroma on the cranial aspect of the distal radial metaphysis causing tenosynovitis of the extensor carpi radialis tendon sheath in a horse.

CASE REPORT: A 3-year-old Thoroughbred gelding was presented with a grade 3/5 lameness of the right forelimb and effusion of the extensor carpi radialis tendon sheath (ECRTS). Radiographic and ultrasonographic examinations revealed an osteochondroma on the cranial aspect of the distal radius projecting into the fibrous lining of the ECRTS. An open surgical approach was used to remove the osteochondroma and some of the proliferative synovial tissue. CONCLUSION: Six months after surgery the clinical signs had resolved and the horse raced successfully.

Autologous bone marrow aspirate for treatment of superficial digital flexor tendonitis in 105 racehorses.

To evaluate a treatment protocol whereby superficial digital flexor (SDF) tendonitis in Thoroughbred and Standardbred racehorses was treated with autologous bone marrow aspirate (ABMA) obtained from the sternebrae. This treatment was combined with desmotomy of the accessory ligament of the SDF tendon (DAL-SDFT) in selected cases. Medical records of 105 horses treated using the reported protocol were reviewed. Signalment, history and details of treatment were recorded. Racing records were reviewed and performance recorded. Of Thoroughbreds, 82 per cent had one or more starts within the follow-up period and 59 per cent had five or more starts. Of Standardbreds, 76 per cent had one or more starts and 62 per cent had five or more starts. A statistically significant difference was found when comparing race starts between sexes, with females having less starts than males (≥1start P=0.017 and ≥5 starts P=0.008, respectively). The proportions of horses having one or more starts and five or more starts did not differ significantly if a DAL-SDFT was performed or not (P=0.31 and 0.63, respectively). Horses with a core lesion in the body of the SDFT have a good prognosis for return to racing following intralesional ABMA injection. Addition of DAL-SDFT to the treatment regimen did not significantly influence outcome.

Use of ultrasound-guided autologous bone marrow transfer for treatment of suspensory ligament desmitis in 30 race horses (2003-2010).

OBJECTIVE: To evaluate the racing performance of horses that underwent ultrasound-guided intralesional injection of autologous bone marrow aspirate for treatment of selected forelimb suspensory ligament (body or branch) core lesions. DESIGN: Retrospective cohort study of 13 Standardbred and 17 Thoroughbred race horses. METHODS: Autologous bone marrow aspirated from the sternebrae was injected, under ultrasound guidance, into suspensory ligament core lesions (body or branch). Racing records were reviewed for a comparison of performance before and after surgery. RESULTS: Of the 13 Standardbreds, 9 (69%) had one or more starts within the follow-up period and 9 (69%) had five or more starts. Of the 17 Thoroughbreds, 15 (88%) had one or more starts within the follow-up period and 12 (71%) had five or more starts. Eight Standardbred horses had at least one start both before and after surgery. Earnings per start did not differ significantly between the three starts immediately after surgery compared with the three starts immediately prior to surgery. Thirteen Thoroughbred horses had at least one start both before and after surgery. Earnings per start were less for the three starts immediately after surgery compared with the three starts immediately prior to surgery. CONCLUSIONS AND CLINICAL RELEVANCE: A horse with a core lesion in the branch or body of the suspensory ligament has a good prognosis for return to racing after treatment with intralesional injection of bone marrow aspirate.

Surgical treatment of septic physitis in 17 foals.

OBJECTIVE: To document the progression and outcome after surgical curettage of septic physes in foals. STUDY DESIGN: Case series of 17 foals, aged <9 weeks, with septic physitis. METHODS: All foals were treated surgically with curettage of physeal lesions. Combinations of systemic antibiotics, intravenous regional perfusion of antibiotic and antibiotic-impregnated polymethylmethacrylate beads were used as adjunct medical treatments. Tissue removed from the physis of three foals during surgery was sent for pathological analysis. RESULTS: At follow-up, 15 of the 17 foals were alive. In the two foals with septic physitis of the distal tibia, the condition was bilateral. Eight foals had concurrent septic arthritis of the joint adjacent to the growth plate, but this did not affect outcome. Referring veterinarians of the foals were contacted at least 9 months post-surgery and all reported excellent functional, and good to excellent cosmetic, outcomes. Pathological examination of the septic physeal tissue showed that normal growth plate had been replaced by inflammatory tissue. CONCLUSION AND CLINICAL RELEVANCE: Septic physitis in foals may be treated using surgical curettage in selected cases. Removal of septic material might be expected to expedite resolution of infection, especially important in immunologically challenged neonates, where septic tissue may act as a reservoir for seeding of infection to other sites.

Distal luxation of the patella in a horse.

A 19-year-old Thoroughbred gelding presented with sudden onset, non-weight bearing lameness in the right hindlimb. Radiography confirmed distal luxation of the patella, which was replaced into its normal anatomical location under general anaesthesia. There were no pathological sequelae noted on follow-up examination 9 months after the initial injury. To our knowledge, this is a rare manifestation of patellar luxation, only reported once previously in the equine literature.

Use of firefly luciferase for ATP measurement: other nucleotides enhance turnover.

Firefly luciferase utilizes only ATP and a few closely related nucleotides as substrates for the formation of luciferyl adenylate which is an intermediate in the bioluminescent reaction sequence that oxidizes firefly luciferin. The enzyme shows two different time courses of light production depending on ATP concentration used: a flash with high concentrations of ATP (> 8 microM) or a fairly constant production of light with lower concentrations of ATP (< 1 microM). Many nucleotides, nucleotide-containing substances and other compounds, when added either prior to or 1 min after the addition of ATP, change the time course of light production. When added before ATP, these compounds yield a reaction mixture in which light production is fairly constant (at the level characteristic of the flash observed with that ATP concentration). When the compounds are added after ATP addition, light production is markedly stimulated and the higher rate of light production is maintained for several minutes. There is an increase in quanta of light produced per luciferase dimer from 1 to 5/min with the addition of any of several nucleotide analogues. These results are consistent with a stimulated release of the inhibitory product oxyluciferin, allowing turnover of the enzyme. This enzyme turnover permits more light output at high ATP concentrations, thus enhancing the sensitivity of enzyme determination.

Intravenous iron administration to very-low-birth-weight newborns receiving total and partial parenteral nutrition.

BACKGROUND: Intravenous iron supplements are not routinely administered to very-low-birth-weight newborns receiving total parenteral nutrition because of the possible increased risk of infection and because iron needs may be met with blood transfusions. METHODS: To assess the benefits of a prudent IV iron supplement (200 to 250 micrograms/kg/d), 26 very-low-birth-weight newborns (birth weight, 1005 +/- 302 g; gestational age, 28 +/- 2.3 weeks; mean +/- SD) were randomly allocated to receive total parenteral nutrition without iron (No-Iron) or with iron supplied as iron dextran (Iron). These newborns were followed at baseline (2 to 3 days after birth) and at weeks 1 to 4 thereafter. At each sampling time, urine samples, fecal samples (rarely), unused total parenteral nutrition solutions, blood products, and a blood sample (1 mL) were collected. RESULTS: There were no differences between the two groups in anthropometric measurements, hematologic or biochemical parameters, number or amount of blood transfusions (2.3 +/- 1.9), amount of blood removed for diagnostic purposes (44 +/- 16 mL), or number of septic events (n = 16). There was no difference between the groups for the total iron excreted; however, the Iron group retained more iron. Iron balance was negative for all but 10 newborns (No-Iron, 3; Iron, 7) throughout the study. CONCLUSIONS: A total iron intake of 400 micrograms/kg/d, half of which was provided by IV iron, is not sufficient to maintain iron balance or to meet fetal accretion rates (1000 micrograms/kg/d) in very-low-birth-weight newborns receiving total parenteral nutrition. Furthermore, endogenous iron from blood transfusions does not provide an adequate supply of iron.

Effect of periodate-oxidized ATP and other nucleotides on firefly luciferase.

Addition of periodate-oxidized ATP (oATP) to firefly luciferase-containing reaction mixtures enhanced light production when the reaction mixture contained > approximately 8 microM ATP. The time course of light production was changed from a flash pattern to a constant light output during incubations of < approximately 10 min. During longer incubation, firefly luciferase was inactivated in a concentration-dependent fashion by oATP. Firefly luciferase has two different time courses of light production that depend on ATP concentration (DeLuca and McElroy, Biochem. Biophys. Res. Commun., 123, 764, 1984). The enhancement of light production occurred only when higher ATP concentrations (> 8 microM) were used. There is little effect of oATP on firefly luciferase activity at low ATP concentrations (< 2 microM) which gave steady production of light. ATP did not antagonize the inactivation of firefly luciferase by oATP. When the oATP was chemically reduced with sodium borohydride (giving or ATP), there was no inactivation of firefly luciferase on incubation. When or ATP was used in a short incubation the enhancement of light production and time course change were the same as those observed with oATP. The corresponding AMP and adenosine compounds (o and or) were slightly inhibitory to firefly luciferase activity. ADP was without effect but both oADP and or ADP enhanced light production. Of these periodate-oxidized ADP, AMP, and adenosine derivatives only oADP inactivated firefly luciferase. The activating effect can be explained by a change in the conformation of the enzyme-product complex so that the product is released faster. In addition there is an inactivation of the enzyme by certain periodate-oxidized nucleotides during longer incubations.

Bioluminescent determination of 0.1 picomole amounts of guanine nucleotides.

A bioluminescence procedure for the determination of the guanylates has been optimized to allow measurement of 0.1 pmol amounts. Modifications of the Karl procedure include the use of purified firefly luciferase and nucleoside diphosphate kinase instead of a crude extract of firefly tails, the use of Tricine buffer instead of the inhibitory arsenate buffer, and optimization of the amounts of reagents and incubation times for each of the partial reactions. In the determination of GMP, background values varied widely with different lots of bovine guanylate kinase. Careful selection of a suitable lot of bovine brain guanylate kinase was essential for determination of lower amounts of guanylates. This establishes that selection of guanylate kinase must be based on experimental determination and not reported adenylate kinase activity. The wide variation in background was not eliminated by the inclusion of adenylate kinase inhibitors.

Enhancement of firefly luciferase activity by cytidine nucleotides.

The temporal pattern of light production by firefly luciferase depends on the ATP concentration. With low concentrations of ATP a constant production of light occurred while at high concentrations of ATP (greater than 10 microM) there was a flash of light followed by a decline in light production. This time course of light production with high ATP concentrations was changed from the flash pattern to a pattern with a constant production of light by several cytidine nucleotides. CTP, CDP, dCTP, dCDP, dideoxyCTP, periodate-oxidized CTP and CDP, and the etheno derivatives of CTP and CDP produced that change. CMP, cytidine, CDP-glycerol, CDP-glucose, CDP-ethanolamine, and benzoylbenzoylCTP either were inhibitory to firefly luciferase or were not effective in changing the flash time course. Coenzyme A and related compounds also changed the time course of light production. The changes in time course produced by either cytidine nucleotides or CoA were inhibited by desulfoCoA. These compounds apparently enhanced light production by promoting the dissociation of the inhibitory product, oxidized luciferin, from the enzyme. When the activating compounds were used with high concentrations of ATP, the sensitivity of assay for firefly luciferase was increased. This increased sensitivity is important when using the firefly luciferase gene as a reporter.

Comparison of properties of commercially available crystalline native and recombinant firefly luciferases.

Commercially available crystalline native and recombinant firefly luciferases were compared. The two types of luciferase had indistinguishable responses to variation in ATP and luciferin concentrations and to omission of reaction components. The time courses of light production, the responses to nucleotide analogues, and the stability of the enzymes under several storage conditions were identical. The native enzyme had a slightly greater specific activity and was more sensitive to trypsin degradation. These differences are probably attributable to differences in conformation.

General practitioners' confidence in diagnosing and managing eye conditions: a survey in south Devon.

In order to open dialogue aimed at increasing eye care in general practice and reduce waiting times for ophthalmic outpatient appointments general practitioners in the Torbay health district were asked about their levels of confidence in ophthalmology as a subject, and in the diagnosis and management of specific eye conditions. They were also asked about ophthalmic equipment available to them, their management policies for 34 specific eye conditions, and their perceived need for further training in this subject. A total of 75% of the general practitioners responded to the questionnaire. Despite more than half of the general practitioners indicating that they did not feel confident with ophthalmology generally, most expressed confidence in diagnosing and managing common eye conditions. Basic equipment for examining the eye was available to most doctors. Referral policies varied considerably, and these have resource implications. Seventy eight per cent of respondents were prepared to take on more eye care in general practice, and over 80% of general practitioners requested informal teaching sessions in ophthalmology. Support must be forthcoming if general practitioners are to provide eye care in general practice.

Peer support group for adolescents with chronic illness.

This study assessed the effects of a monthly peer support group for adolescents with cancer and other hematological diseases. These adolescents shared activities and experiences with nondisabled high school students. At the group's conclusion, the adolescents reported that the group helped them cope with their illness and improved the quality of their daily lives. Nondisabled students reported that the group favorably affected their attitudes about, and intended behavior toward, peers with chronic illnesses. These results suggest that such groups can provide important benefits for individuals with chronic illnesses as well as for their nondisabled peers.

The Exeter Diabetic Project: an acceptable district-wide education programme for general practitioners.

The Health District is a logical administrative unit for planning health services, and therefore the feasibility of delivering education for general practitioners across one such area was studied. By providing a suitable educational programme at convenient locations, it was possible to involve 89% out of 180 principals in General Practice in the Exeter Health District. Two-thirds of these general practitioners attended educational sessions away from the District Postgraduate Centre. In a questionnaire answered by 81% of the involved general practitioners, 98% acknowledged responsibility for managing their diabetic patients, but 76% admitted to inadequate skills in retinal assessment. In a follow-up postal questionnaire answered by 64% of general practitioners involved in the project, 74% found that they learnt from the retinopathy screening clinic and 100% found the sessions on eye disease helpful. The education programme was designed to link with a District-wide retinopathy screening initiative.

Evaluation of virus neutralization antibody levels in diabetic mutant mice.

Experiments were designed to determine whether genetic predisposition to diabetes or overt diabetes in the identical genotype had an effect on the level of neutralization antibodies to coxsackievirus B4 (CB4). Quantitation of neutralization antibody (NT) levels against CB4 was performed using both the 50% endpoint procedure and the multivariate Wilcoxon rank sum test. The results of these experiments demonstrate that the use of the multivariate procedure for the analysis of neutralization antibody levels provides both quantitative and qualitative information not evident when only the classical 50% end point procedure is used. Moreover, when comparison on antibody levels between different groups is being considered, the power of the comparison is markedly greater using the multivariate Wilcoxon rank sum test results. The present report provide an illustration of the difference in the qualitative and quantitative information obtained by the end-point procedure and the more comprehensive multivariate procedure for the analysis of neutralization antibody levels in diabetic mutant mice infected with coxsackievirus B4.