Delivery of growth factors using gene therapy to enhance bone healing

OBJECTIVE: To review the delivery of growth factors using gene therapy for enhancing long-bone fracture healing. STUDY DESIGN: Literature review. METHODS: MEDLINE and CAB Abstracts literature search (1980-2004). RESULTS: Non-union and infected non-union are relatively common complications of long-bone fractures in human and veterinary patients. Growth factors are cytokines that regulate many cell functions important in fracture healing. Exogenous growth factors can be delivered to the fracture site as recombinant proteins or using gene therapy. Recombinant human bone morphogenetic protein-2 and -7 (rhBMP-2 and -7), in particular, enhance fracture healing in numerous experimental and clinical studies. Some limitations with use of recombinant proteins may be overcome by use of gene therapy. Gene therapy involves delivery of the growth factor gene to cells at the fracture site using a viral or non-viral vector. The gene is then expressed (protein synthesis) by cells at the fracture site. Delivery of the BMP gene to the fracture site using gene therapy has been evaluated in laboratory animal models of non-union, with favorable results and without complications. CONCLUSION: Delivery of growth factors, particularly members of BMP family, to the fracture site using gene therapy may be a method to enhance fracture healing. Use of this technology may improve the prognosis for patients with long-bone fractures. CLINICAL RELEVANCE: Clinical application of gene therapy could improve the prognosis for human and veterinary patients with long-bone fractures, but has not been evaluated clinically.     

Use of a semitendinosus myocutaneous flap for soft-tissue reconstruction of a grade IIIB open tibial fracture in a dog

OBJECTIVE: To describe the clinical use of a semitendinosus myocutaneous flap for soft-tissue reconstruction of a grade IIIB open tibial fracture. STUDY DESIGN: Case report. ANIMALS: A 5-year-old castrated male Labrador retriever with a grade IIIB open tibial fracture. METHODS: A myocutaneous flap was created by elevating the origin of the semitendinosus muscle and the associated overlying skin. The flap was rotated distally based on the distal vascular pedicle into a soft-tissue defect overlying the central and distal third of the tibia. RESULTS: The flap allowed one-stage reconstruction of a severe soft-tissue defect overlying denuded bone in a grade IIIB open tibial fracture. The muscular portion of the flap survived; however, there was necrosis of 3 cm of skin extending distally beyond the muscular portion of the flap. CONCLUSION: Distal rotation of the semitendinosus muscle and overlying skin can be used in the treatment of severe soft-tissue defects of the canine crus. CLINICAL RELEVANCE: Complex soft-tissue injuries to the canine crus can be treated with a myocutaneous flap as a reasonable alternative to other reconstructive techniques.     

Biomechanical comparison of cortical radial graft versus ulnar transposition graft limb-sparing techniques for the distal radial site in dogs

OBJECTIVE: To compare the biomechanical properties of radial graft (RG) versus ulnar transposition graft (UTG) limb-sparing techniques in the dog. STUDY DESIGN: Cadaveric study. ANIMALS: Six pairs of normal canine thoracic limbs. METHODS: In each pair of limbs, 1 limb was subjected to the RG technique and the other to the UTG technique. Limbs were tested in axial loading until failure. Modes of failure and biomechanical properties were compared between the 2 groups. Percent coverage of the metacarpal bone by the plate was retrospectively compared between the limbs that failed by fracture of the metacarpal bones and those that did not. RESULTS: RG limbs had significantly greater stiffness, yield load, maximum load, maximum energy, and post-yield energy. All UTG limbs failed by cranial bending of the plate. Half of the RG limbs failed by caudal bending of the plate and half by fracture of the third metacarpal bone at the distal end of the plate. Limbs with <80% plate coverage of the metacarpal bone were significantly more likely to fail by metacarpal fracture. CONCLUSIONS: The RG technique was biomechanically superior to the UTG technique under the conditions studied. At least 80% plate coverage of the metacarpal bone should be achieved. CLINICAL RELEVANCE: Exercise restriction and coaptation bandages should be considered for patients undergoing the UTG technique to minimize potential biomechanical complications.     

Cardiac troponin-I concentration in dogs with cardiac disease

Cardiac troponin-I (cTnI) is a highly sensitive and specific marker of myocardial injury and can be detected in plasma by immunoassay techniques. The purpose of this study was to establish a reference range for plasma cTnI in a population of healthy dogs using a human immunoassay system and to determine whether plasma cTnI concentrations were high in dogs with acquired or congenital heart disease, specifically cardiomyopathy (CM), degenerative mitral valve disease (MVD), and subvalvular aortic stenosis (SAS). In total, 269 dogs were examined by physical examination, electrocardiography, echocardiography, and plasma cTnI assay. In 176 healthy dogs, median cTnI was 0.03 ng/mL (upper 95th percentile = 0.11 ng/mL). Compared with the healthy population, median plasma cTnI was increased in dogs with CM (0.14 ng/mL; range, 0.03-1.88 ng/mL; P < .001; n = 26), in dogs with MVD (0.11 ng/mL; range, 0.01-9.53 ng/mL; P < .001; n = 37), and in dogs with SAS (0.08 ng/mL; range, 0.01-0.94 ng/mL; P < .001; n = 30). In dogs with CM and MVD, plasma cTnI was correlated with left ventricular and left atrial size. In dogs with SAS, cTnI demonstrated a modest correlation with ventricular wall thickness. In dogs with CM, the median survival time of those with cTnI >0.20 ng/mL was significantly shorter than median survival time of those with cTnI <0.20 ng/mL (112 days versus 357 days; P = .006). Plasma cTnI is high in dogs with cardiac disease, correlates with heart size and survival, and can be used as a blood-based biomarker of cardiac disease.     

Echocardiographic estimation of mean left atrial pressure in a canine model of acute mitral valve insufficiency

High mean left atrial pressure (MLAP) due to canine degenerative mitral valve disease is associated with clinically relevant morbidity and mortality. The ability to noninvasively measure MLAP would assist in the diagnosis and treatment of disease. Doppler echocardiography allows measurement of early transmitral blood flow (E) and the velocity of the mitral valve annulus (Ea). The ratio of early mitral inflow velocity to early mitral annular velocity (E: Ea) correlates well with MLAP in human subjects. We sought to determine the ability of E: Ea to predict MLAP in dogs with experimentally induced mitral regurgitation. Nine anesthetized purpose-bred dogs underwent placement of a Swan-Ganz catheter into the left atrium and recording of MLAP. Simultaneous transthoracic echocardiographic and hemodynamic studies were performed after acute chordae tendineae rupture and during IV infusion with nitroprusside (2.5-5.0 microg x kg(-1) x min(-1)) or hydralazine (1-1.5 mg/kg). Mitral regurgitant fraction, measured by single-plane angiography and thermodilution, ranged from 17% to 81%. MLAP increased from 5.4 +/- 2.5 mm Hg to 17.4 +/- 9.4 mm Hg after creation of mitral valve regurgitation (MR; P = .018). Forty sets of echocardiographic measurements were obtained from 7 dogs, and E, as well as E: Ea, were linearly related to MLAP. The R2 value for the linear regression equation containing E: Ea as the dependent variable (0.83) was greater than that for E (0.73). The 95% confidence intervals were calculated for predicting MLAP = 20 mm Hg from E:Ea, and E:Ea >9.1 or <6.0 indicated a 95% probability that MLAP was >20 mm Hg or <20 mm Hg, respectively. Echocardiography can be used to predict MLAP in isoflurane-anesthetized dogs with experimentally induced acute mitral valve insufficiency.     

Exhaled breath condensate (EBC) collection in cats--description of a non-invasive technique to investigate airway disease

Exhaled breath condensate has been collected in other species and used as a non-invasive method of evaluating airway disease by measurement of various markers in the fluid, including hydrogen peroxide, nitric oxide, leukotrienes and prostaglandins. We describe a novel technique for the collection of exhaled breath condensate from cats, which enabled collection of fluid and measurement of its hydrogen peroxide concentration. Further studies will be needed to establish the value of this technique in the investigation of feline respiratory disease.     

Use of imidacloprid-permethrin to prevent transmission of Anaplasma phagocytophilum from naturally infected Ixodes scapularis ticks to dogs

One group of eight beagles was treated with a combination of imidacloprid and permethrin 7 days before exposure to Ixodes scapularis ticks that were naturally infected with Anaplasma phagocytophilum. A second group of eight beagles was not treated and was also exposed to infected ticks. Seven of eight non-treated dogs--but none of the treated dogs--developed specific antibodies to A. phagocytophilum. Results of this study indicate that a combination of imidacloprid and permethrin can prevent transmission of A. phagocytophilum to dogs if administered before exposure to infected ticks.     

Influence of ethylene action, storage atmosphere, and storage duration on diphenylamine and diphenylamine derivative content of Granny Smith apple peel

The application of diphenylamine (DPA) to prevent the apple peel disorder superficial scald can result in accumulation of a number of DPA derivatives resulting from C-nitration, C-hydroxylation, O-methylation, and N-nitrosation during fruit storage. As the presence of these compounds may be indicative of metabolic processes leading to superficial scald development, the contents of DPA and DPA derivatives were determined in fruits treated at harvest with DPA or DPA plus the ethylene action inhibitor 1-methylcyclopropene (1-MCP), which also prevents scald development. Influences of fruit maturity, storage environment, storage duration, and a 14 day poststorage ripening period on accumulation of DPA metabolites were also assessed. Poststorage ripening, 1-MCP treatment, and controlled atmosphere storage had varied effects on DPA derivative contents suggesting that reactive oxygen and nitrogen species, such as *OH, *NO, and *NO2, or enzyme-catalyzed reactions may be present during certain ripening and senescence-related physiological processes. Definitive correlations between superficial scald incidence and contents of specific derivatives were not observed.     

Tim50 maintains the permeability barrier of the mitochondrial inner membrane

Transport of metabolites across the mitochondrial inner membrane is highly selective, thereby maintaining the electrochemical proton gradient that functions as the main driving force for cellular adenosine triphosphate synthesis. Mitochondria import many preproteins via the presequence translocase of the inner membrane. However, the reconstituted Tim23 protein constitutes a pore remaining mainly in its open form, a state that would be deleterious in organello. We found that the intermembrane space domain of Tim50 induced the Tim23 channel to close. Presequences overcame this effect and activated the channel for translocation. Thus, the hydrophilic cis domain of Tim50 maintains the permeability barrier of mitochondria by closing the translocation pore in a presequence-regulated manner.     

The breakup of a main-belt asteroid 450 thousand years ago

Collisions in the asteroid belt frequently lead to catastrophic breakups, where more than half of the target's mass is ejected into space. Several dozen large asteroids have been disrupted by impacts over the past several billion years. These impact events have produced groups of fragments with similar orbits called asteroid families. Here we report the discovery of a very young asteroid family around the object 1270 Datura. Our work takes advantage of a method for identification of recent breakups in the asteroid belt using catalogs of osculating (i.e., instantaneous) asteroid orbits. The very young families show up in these catalogs as clusters in a five-dimensional space of osculating orbital elements.