Forced unfolding of proteins within cells

To identify cytoskeletal proteins that change conformation or assembly within stressed cells, in situ labeling of sterically shielded cysteines with fluorophores was analyzed by fluorescence imaging, quantitative mass spectrometry, and sequential two-dye labeling. Within red blood cells, shotgun labeling showed that shielded cysteines in the two isoforms of the cytoskeletal protein spectrin were increasingly labeled as a function of shear stress and time, indicative of forced unfolding of specific domains. Within mesenchymal stem cells-as a prototypical adherent cell-nonmuscle myosin IIA and vimentin are just two of the cytoskeletal proteins identified that show differential labeling in tensed versus drug-relaxed cells. Cysteine labeling of proteins within live cells can thus be used to fluorescently map out sites of molecular-scale deformation, and the results also suggest means to colocalize signaling events such as phosphorylation with forced unfolding.     

Treatment of equine glaucoma by transscleral neodymium:yttrium aluminum garnet laser cyclophotocoagulation: a retrospective study of 23 eyes of 16 horses

Contact neodymium:yttrium aluminum garnet (Nd:YAG) laser transscleral cyclophotocoagulation (TSCP) was performed on 23 eyes of 16 horses for treatment of glaucoma. The mean highest preoperative IOP was 51 ± 17 mmHg. Follow-up evaluation was available for 19 eyes 1 day after surgery, 14 eyes from 1 to 2 weeks, 16 eyes from 4 to 6 weeks, 9 eyes from 12 to 16 weeks, and 10 eyes greater than 20 weeks after laser treatment. The mean intraocular pressure (IOP) the day following surgery was 34 ± 13 mmHg. The mean IOP for each follow-up period was: one to two weeks postoperative, 23 ± 9 mmHg; four to six weeks, 24 ± 7 mmHg; 12–16 weeks, 28 ± 10 mmHg; and  20 weeks, 22 ± 9 mmHg. IOP measurements were significantly different from pretreatment values for all follow-up intervals except for weeks 12–16 ( P < 0.05). Treatment success was defined as maintenance of IOP < 30 mmHg. Treatment success was achieved in 93%, 88%, 78%, and 70% of the treated eyes at the 1–2 weeks, 4–6 weeks, 12–16 weeks, and the  20 weeks re-evaluation, respectively. No significant difference was found between the number of eyes visual at presentation (52.2%) and visual at 20 weeks (60%). The most common laser complications were conjunctival hyperemia (21.7%) and corneal ulcers (13.0%). Results of this study indicate that Nd:YAG TSCP is an effective method of controlling IOP and preserving vision in horses with glaucoma. An effective Nd:YAG laser protocol for TSCP in the equine glaucomatous eye is a power setting of 11 W, duration of 0.4 s, applied 5 mm posterior to the limbus at 60 sites, resulting in a total energy dose of 264 J.     

Synovial myxoma: magnetic resonance imaging in the assessment of an unusual canine soft tissue tumour

Magnetic resonance imaging (MRI) of the hindlimb of a 10-year-old Labrador retriever was performed preoperatively to define the limits and invasive nature of a synovial myxoma. This unusual tumour in dogs has also only rarely been reported in humans, although the use of advanced imaging techniques has been more widely reported in the assessment of soft tissue tumours in people. MRI was an invaluable aid in the delineation of the extensive pathological changes associated with this tumour and consequently its surgical treatment. Amputation was performed and the dog remained disease-free 18 months after surgery.     

Monoclonal Gammopathy in a Dog With Chronic Pyoderma

A monoclonal gammopathy composed of immunoglobulin G, with concurrent light-chain proteinuria and generalized lymph node plasmacytosis, was associated with chronic pyoderma in a dog. A uniform population of plasma cells was observed cytologically and histologically in multiple lymph node specimens. A diagnosis of monoclonal gammopathy of unknown significance was eventually made by exclusion of other known causes of monoclonal gammopathy, resolution after antibiotic therapy, and no evidence of lymphoproliferative disease after 11 months of follow-up and subsequent necropsy. This report expands the diagnostic considerations for monoclonal gammopathies in the dog.     

Evaluation of various compounds to inhibit activity of matrix metalloproteinases in the tear film of horses with ulcerative keratitis

OBJECTIVE: To examine in vitro effects of various antiproteolytic compounds on activity of matrix metalloproteinase (MMP)-2 and -9 in the tear film of horses with active corneal ulcers. SAMPLE POPULATION: Samples of tear film obtained from the eyes of 34 horses with active ulcerative keratitis. PROCEDURE: Horses were sedated, and tear samples were collected from the lower fornix of 34 ulcerated eyes by use of capillary tubes. The protease inhibitors 0.2% EDTA, 0.1% doxycycline, 10% N-acetylcysteine (NAC), 0.1% solution of a modified dipeptide that contains hydroxamic acid (ie, ilomostat), 0.1% alpha1-proteinase inhibitor (PI), 0.5% alpha1-PI, and 100% fresh equine serum (ES) were used to treat pooled samples. Amount of latent and active MMP-2 and -9 was measured by optical density scanning of gelatin zymograms of treated and untreated tear samples. RESULTS: Pooled tear samples obtained from ulcerated eyes contained the latent and active forms of MMP-2 and -9. Compared with MMP activity in untreated samples, total MMP activity (sum of all bands detected) observed on the gelatin zymogram gels was reduced by 99.4% by EDTA, 96.3% by doxycycline, 98.8% by NAC, 98.9% by ilomostat, 52.4% by 0.1% alpha1-PI, 93.6% by 0.5% alpha1-PI, and 90.0% by ES. CONCLUSIONS AND CLINICAL RELEVANCE: We documented that EDTA, doxycycline, NAC, ilomostat, alpha1PI, and ES inhibited MMP activity in vitro. Because these compounds use different mechanisms to inhibit various families of proteases in the tear film of horses, a combination of these protease inhibitors may be beneficial for treatment of corneal ulcers in horses.     

Effect of anticonvulsant dosages of potassium bromide on thyroid function and morphology in dogs

A placebo-controlled experiment was performed to evaluate the effect of potassium bromide on the canine thyroid gland. Basal total thyroxine, free thyroxine, and basal thyrotropin serum concentrations were evaluated over a 6-month period in potassium bromide-treated and control dogs. A thyrotropin-releasing hormone stimulation test was also performed in all dogs at the beginning and conclusion of the study. Thyroid histopathology was compared between treated and control dogs at the end of the study. No difference was detected in any parameter between the two groups at the end of the study. A decline in thyroid hormone concentrations over the course of the study did occur in both groups of dogs. Potassium bromide does not appear to have a significant effect on canine thyroid function or morphology.     

The accessory lung lobe in thoracic disease: a case series and anatomical review

Diseases of the accessory lung lobe (AccLL) are often overlooked in dogs and cats, perhaps because of its recessed location. Diseases of this lobe also can mimic diseases of the diaphragm or structures comprising the caudal mediastinum. Normal anatomy of the AccLL and the diagnostic tools used today will be reviewed to help differentiate disease of adjacent organs. Four cases will also be described to illustrate disease presentation and radiographic findings affecting this lobe.