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Effects of physiologic concentrations of l-lysine on in vitro replication of feline herpesvirus 1 Texte intégral
2014
Cave, Nicholas J. | Dennis, Kathryn | Gopakumar, Gaya | Dunowska, Magda
Objective-To evaluate the effects of various concentrations of l-lysine on in vitro replication of feline herpesvirus 1 (FHV-1). Sample- Cultures of Crandell-Rees feline kidney (CRFK) cells. Procedures- CRFK cells were inoculated with FHV-1 and maintained in media with 20 combinations of l-arginine and l-lysine concentrations. Changes in cell viability were monitored by continuous measurement of electrical impedance of cultured cells and by observation of viral cytopathic effects. Viral load was determined by use of quantitative PCR assay in supernatants obtained from infected cultures at specified time points. Results- Increases in l-lysine concentration had no effect on the kinetics of cell death in FHV-1-infected cultures. There was also no significant effect (r2 < 0.1) on viral DNA load for l-arginine concentrations ≥ 12 μg/mL There was a significant effect of increases in l-lysine concentration on viral DNA load in media supplemented with 6 μg of l-arginine/mL (mean ± SD slope, −4,641 ± 1,626 units; adjusted r2 = 0.45). However, the difference between the lowest (1 × 10(6.28) copies/μL) and highest (1 × 10(6.86) copies/μL) FHV-1 DNA load in these media was < 1 logarithm. Conclusions and Clinical Relevance-The difference in FHV-1 DNA load was unlikely to be biologically important. Various l-lysine concentrations did not inhibit in vitro replication of FHV-1 at l-arginine concentrations sufficient to maintain cell growth. This conclusion was consistent with results of other studies in which investigators have not detected a consistently beneficial effect when l-lysine is administered to FHV-1-infected cats.
Afficher plus [+] Moins [-]Detection of misfolded prion protein in retina samples of sheep and cattle by use of a commercially available enzyme immunoassay Texte intégral
2014
Smith, Jodi D. | Greenlee, Justin J.
Objective-To determine the usefulness of retina samples for detection of disease-associated prion protein by use of a commercially available enzyme immunoassay (EIA) intended for rapid identification of sheep and cattle with transmissible spongiform encephalopathies (TSEs). Samples-Retina, brainstem at the level of the obex, and retropharyngeal lymph node samples obtained from 15 TSE-inoculated sheep (scrapie [n = 13] or transmissible mink encephalopathy passaged through a bovid [2]); retina and brainstem samples obtained from 11 TSE-inoculated cattle (transmissible mink encephalopathy passaged through a bovid [7] or classical BSE [4]); and negative control tissue samples obtained from 2 sheep and 2 cattle that were not inoculated with TSEs. Procedures-Tissue samples were homogenized and analyzed for detection of abnormally folded disease-associated prion protein with a commercially available EIA and 2 confirmatory assays (western blot analysis or immunohistochemical analysis). Results-Retina sample EIA results were in agreement with results of brainstem sample EIA or confirmatory assay results for negative control animals and TSE-inoculated animals with clinical signs of disease. However, TSE-inoculated animals with positive confirmatory assay results that did not have clinical signs of disease had negative retina sample EIA results. Retina sample EIA results were in agreement with brainstem sample immunohistochemical results for 4 TSE-inoculated sheep with negative retropharyngeal lymph node EIA results. Conclusions and Clinical Relevance-Results of this study suggested that retina samples may be useful for rapid EIA screening of animals with neurologic signs to detect TSEs.
Afficher plus [+] Moins [-]Assessment of folate receptor expression and folate uptake in multicentric lymphomas in dogs Texte intégral
2014
Objective-To determine expression of folate receptors (FRs) and folate uptake in multicentric lymphomas in dogs. Sample-10 dogs with histopathologically confirmed multicentric lymphoma and 20 archival lymph node biopsy specimens from dogs with multicentric lymphoma. Procedures-Multicentric lymphomas in 10 dogs were prospectively evaluated for FR expression by use of immunohistochemical analysis and for in vivo folate uptake by use of nuclear scintigraphy. Dogs with FR-expressing tumors were eligible for FR-targeted chemotherapy. Twenty archival lymphoma biopsy specimens were also evaluated with immunohistochemical analysis. Results-FRs were not detected with immunohistochemical analysis in lymph node samples obtained from the 10 dogs or in archival biopsy specimens. However, nuclear scintigraphy revealed uptake of radioactive tracer in 6 of 10 dogs. Five of these 6 dogs were treated with an FR-targeted chemotherapeutic agent; results of treatment were complete remission in 1 dog, stable disease in 2 dogs, and progressive disease in 2 dogs. Treatment-related toxicoses generally were mild. Conclusions and Clinical Relevance-This study provided strong evidence for folate uptake in a substantial portion of multicentric lymphomas of dogs and indicated the antitumor activity of FR-targeted chemotherapeutics for these cancers. Use of FR-targeted chemotherapeutics may be promising for the treatment of FR-expressing multicentric lymphomas in dogs. Further studies are needed to determine reasons for lack of immunoreactivity to currently identified anti-FR antibodies and to develop improved methods for detecting FRs in lymphomas of dogs.
Afficher plus [+] Moins [-]In vitro evaluation of anatomic landmarks for the placement of suture to achieve effective arytenoid cartilage abduction by means of unilateral cricoarytenoid lateralization in dogs Texte intégral
2014
Objective- To evaluate anatomic landmarks to define the ideal suture placement location to achieve appropriate and consistent arytenoid cartilage abduction via unilateral cricoarytenoid lateralization (UCL) in dogs. Sample- 6 cadaveric canine larynges. Procedures- Laryngeal airway resistance (LAR) was determined for each specimen before (baseline) and after suture placements with the epiglottis open and closed. To achieve UCL, suture was placed through the cricoid cartilage just caudal to the cricoarytenoid articulation (suture placement position [SPP] 1), one-fourth of the distance caudally between the cricoarytenoid and cricothyroid articulations (SPP 2), and three-fourths of the distance caudally between the cricoarytenoid and cricothyroid articulations (SPP 3). The LAR was again calculated after tensioning of each suture separately. Results- With a closed epiglottis, median LAR was 30.0, 20.4, 11.4, and 3.3 cm H2O/L/s at baseline and SPPs 1, 2, and 3, respectively. After UCL at SPP 1, LAR with the epiglottis closed was not significantly different from that at baseline. With an open epiglottis, median LAR was 2.0, 0.4, 0.2, and 0.0 cm H(2)O/L/s at baseline and SPPs 1, 2, and 3, respectively. After UCL at SPPs 1, 2, or 3, LAR with an open epiglottis was significantly lower than that at baseline. Conclusions and Clinical Relevance- Results indicated that placement of suture through the cricoid cartilage at the caudal border of the cricoarytenoid articulation was appropriate to sufficiently reduce LAR without increasing the risk of aspiration pneumonia through overabduction of the arytenoid cartilage.
Afficher plus [+] Moins [-]Expression of epidermal growth factor receptor and human epidermal growth factor receptor 2 in periocular squamous cell carcinomas of horses Texte intégral
2014
Hendrix, Diane V. H. | Newkirk, Kim M.
Objective—To determine whether epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 (HER2) are expressed in periocular squamous cell carcinomas (SCCs) of horses. Sample—Biopsy specimens of SCCs from 46 horses. Procedures—Pathology records were searched retrospectively for biopsy specimens of periocular SCCs obtained from horses. Slides of the specimens were reviewed histologically to confirm the SCC diagnosis and stained for EGFR and HER2 by immunohistochemical methods. For both EGFR and HER2, the immunohistochemical staining intensity and percentage of stain-positive cells on the slides were determined. Results—43 of 46 (93%) SCCs were immunoreactive for EGFR. The median score for EGFR staining intensity was 4 (range, 2 to 12), and the median number of mitotic figures was 8 mitotic figures/10 hpfs (range, 0 to 34 mitotic figures/10 hpfs). Mitotic index was not correlated with the percentage of EGFR stain–positive cells or staining intensity. Of the 43 EGFR-immunoreactive SCCs, 38 had stain present primarily in the cytoplasm and 5 had stain equally distributed between the cytoplasm and cell membranes. Thirty-five of 46 (76%) SCCs were immunoreactive for HER2. Mitotic index was not correlated with the percentage of HER2 stain–positive cells or staining intensity. Of the 35 HER2-immunoreactive SCCs, the stain was present primarily in the cytoplasm and 7 had stain equally distributed between the cytoplasm and cell membranes. Conclusions and Clinical Relevance—Results indicated that most periocular SCCs of horses expressed EGFR and HER2, which suggested that equine periocular SCCs might respond to treatment with EGFR inhibitors.
Afficher plus [+] Moins [-]Evaluation of tissue oxygen saturation with near-infrared spectroscopy during experimental acute hemorrhagic shock and resuscitation in dogs Texte intégral
2014
Pavlisko, Noah D. | Henao-Guerrero, Natalia | Killos, Maria B. | Ricco, Carolina | Shih, Andre C. | Bandt, Carsten | Werre, Stephen R.
Objective-To evaluate tissue oxygen saturation (Sto2) by use of near-infrared spectroscopy in experimental acute hemorrhagic shock and resuscitation in dogs. Animals-14 healthy adult purpose-bred Beagles. Procedures-Dogs were anesthetized with isoflurane via facemask, anesthesia was maintained with propofol and rocuronium bromide, and dogs were mechanically ventilated to maintain normocapnia. Dogs were studied under normovolemia (baseline), hypovolemia with target mean arterial blood pressure < 40 mm Hg achieved and maintained steady for 10 minutes (hypovolemia T1), then 20 minutes later (hypovolemia T2), following resuscitation with shed blood (after transfusion), and after administration of 20 mL of hetastarch/kg (hypervolemia). Conditions were executed sequentially during a single anesthetic episode, allowing stabilization between states (10 minutes). Hemoglobin concentration, mean arterial blood pressure, arterial blood gas concentrations, cardiac index, oxygen delivery indexed to body surface area, and Sto2 were monitored. Results-From baseline to hypovolemia T1, there was a significant reduction in mean +/- SD oxygen delivery index (619 ± 257 mL/min/m2 to 205 ± 76 mL/min/m2) and StO2 (94 ± 4.4% to 78 ±12.2%). Following resuscitation, Sto2 (80 ± 8.5% vs 92 ± 6.45%) and oxygen delivery index (211 ± 73 mL/min/m2 vs 717 ± 221 mL/min/m2) significantly increased, returning to baseline values. Hypervolemia had no effect on Sto2 or oxygen delivery index. A strong correlation (r = 0.97) was detected between mean oxygen delivery index and Sto2 across all time points. Conclusions and Clinical Relevance-Under the conditions of this study, there was a strong correlation between Sto2 and oxygen delivery, suggesting that Sto2 may be used to estimate oxygen delivery.
Afficher plus [+] Moins [-]Characterization of IgE-mediated cutaneous immediate and late-phase reactions in nonallergic horses Texte intégral
2014
Woodward, Michelle C. | Andrews, Frank M. | Kearney, Michael T. | Del Piero, Fabio | Hammerberg, Bruce | Pucheu-Haston, Cherie M.
Objective—To characterize the response of skin of nonallergic horses following ID injection of polyclonal rabbit anti-canine IgE (anti-IgE) and rabbit IgG. Animals—6 healthy horses. Procedures—Skin in the cervical area was injected ID with anti-IgE and IgG. Wheal measurements and skin biopsy specimens were obtained before and 20 minutes and 6, 24, and 48 hours after injection. Tissue sections were evaluated for inflammatory cells at 4 dermal depths. Immunohistochemical analysis for CD3, CD4, and CD8 was performed, and cell counts were evaluated. Results—Anti-IgE wheals were significantly larger than IgG wheals at 20 minutes and 6 and 24 hours after injection. There were significantly more degranulated mast cells after anti-IgE injection than after IgG injection. There were significantly more eosinophils at 6, 24, and 48 hours and neutrophils at 6 hours after anti-IgE injection, compared with cell numbers at those same times after IgG injection. There were significantly more eosinophils in the deeper dermis of anti-IgE samples, compared with results for IgG samples. No significant differences between treatments were detected for CD3+, CD4+, or CD8+ cells. Conclusions and Clinical Relevance—Injection of anti-IgE antibodies was associated with the development of gross and microscopic inflammation characterized by mast cell degranulation and accumulation of inflammatory cells, particularly eosinophils and neutrophils. This pattern appeared to be similar to that of horses with naturally developing allergic skin disease, although lymphocytes were not increased; thus, ID injection of anti-IgE in horses may be of use for evaluating allergic skin diseases of horses.
Afficher plus [+] Moins [-]Acute exercise does not induce an acute phase response (APR) in Standardbred trotters Texte intégral
2014
Kristensen, Lena | Buhl, Rikke | Nostell, Katarina | Bak, Lars | Petersen, Ellen | Lindholm, Maria | Jacobsen, Stine
The purpose of the study was to investigate whether acute strenuous exercise (1600- to 2500-m race) would elicit an acute phase response (APR) in Standardbred trotters. Blood levels of several inflammatory markers [serum amyloid A (SAA), haptoglobin, fibrinogen, white blood cell count (WBC), and iron], muscle enzymes [creatinine kinase (CK) and aspartate transaminase (AST)], and hemoglobin were assessed in 58 Standardbred trotters before and after racing. Hemoglobin levels increased and iron levels decreased 12 to 14 h after racing and haptoglobin concentrations, white blood cell counts, and iron levels were decreased 2 and/or 7 d after racing. Concentrations of CK, AST, SAA, and fibrinogen were unaltered in response to racing. Acute strenuous exercise did not elicit an acute phase reaction. The observed acute increase in hemoglobin levels and decreases in haptoglobin and iron levels may have been caused by exercise-induced hemolysis, which indicates that horses might experience a condition similar to athlete’s anemia in humans. The pathogenesis and clinical implications of the hematological and blood-biochemical changes elicited by acute exercise in Standardbred trotters in the present study warrant further investigation.
Afficher plus [+] Moins [-]Comparison of an antigen-capture enzyme-linked immunosorbent assay with bacterial culture for detection of Salmonella in poultry-hatchery environmental samples Texte intégral
2014
Brooks, B.W. | Lutze-Wallace, C.L. | Devenish, J. | Elmufti, M. | Paris, Jethro
An antigen-capture, monoclonal-antibody-based enzyme-linked immunoassay (ELISA) that detects a broad range of Salmonella serovars in various serogroups was developed and compared with standard culture procedures for detection of Salmonella in 1055 field samples collected from poultry-hatchery environments. The diagnostic sensitivity of the ELISA relative to culture was 99.9% and the diagnostic specificity 99.6%. The extensive culture procedure included nonselective enrichment (NSE) as well as primary selective enrichment (PSE) and delayed secondary enrichment (DSE) with Hajna tetrathionate (TT) and Rappaport-Vassiliadis (RV) selective-enrichment broths. Significantly more Salmonella-positive samples were detected by ELISA and culture at the DSE stage than at the NSE and PSE stages (P < 0.05). Significantly more RV than TT broths were positive for Salmonella by culture and ELISA by the DSE stage (P < 0.05). This ELISA procedure could be a reliable screening test for the detection of Salmonella in hatchery samples.
Afficher plus [+] Moins [-]Effects of meloxicam and phenylbutazone on renal responses to furosemide, dobutamine, and exercise in horses Texte intégral
2014
Raidal, Sharanne L. | Hughes, Kris J. | Charman, Amanda-Lee | Nielsen, Sharon G. | Phillips, Jaqueline K. | Noble, Glenys K.
Objective—To compare the effects of 2 NSAIDs (phenylbutazone and meloxicam) on renal function in horses. Animals—9 Thoroughbred or Standardbred mares (mean ± SD age, 5.22 ± 1.09 years [range, 2 to 12 years]; mean body weight, 470 ± 25 kg [range, 442 to 510 kg]). Procedures—A randomized blinded placebo-controlled crossover study was conducted to examine the effects of treatment with phenylbutazone, meloxicam, or a placebo (control solution) on renal responses to the administration of furosemide, dobutamine, and exercise (15 minutes at 60% of maximum heart rate). Renal function was assessed by use of bilateral ureteral catheterization for simultaneous determination of creatinine clearance, sodium excretion, and urine flow rate. Results—Both phenylbutazone and meloxicam attenuated diuresis and natriuresis and reduced glomerular filtration rate, compared with results for the control solution, when horses were treated with furosemide. Mean arterial blood pressure, urine flow rate, and glomerular filtration rate were increased during or after (or both) dobutamine infusion. Both NSAIDs reduced urine flow rate and sodium excretion associated with dobutamine infusion and exercise but had no effect on glomerular filtration rate. Conclusions and Clinical Relevance—Responses to meloxicam, a cyclooxygenase (COX)-2 preferential agent, appeared comparable to those detected after phenylbutazone treatment, which suggested that COX-2 was the mediator of prostanoid-induced changes to renal function in horses and indicated that COX-2–preferential agents would be likely to have adverse renal effects similar to those for nonselective COX inhibitors in volume-depleted horses.
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