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  • Author or Editor: John P. Loftus x
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Abstract

Objective—To assess expression pattern and subcellular compartmentalization of 5-lipoxygenase in cutaneous, UV radiation–induced, and oral squamous cell carcinomas (SCCs) in cats and determine the effects of cyclooxygenase or 5-lipoxygenase inhibition on proliferation or apoptosis in a feline oral squamous cell carcinoma (SCCF1) cell line.

Sample—60 archived paraffin-embedded samples of SCCs from 60 cats and SCCF1 cells.

Procedures—Retrospective immunohistochemical analysis of the archived samples of SCCs (20 cutaneous, 20 UV radiation–induced, and 20 oral tumors) was performed. Cell culture proliferation assays involving SCCF1 cells were performed, and tepoxalin-induced apoptosis and signaling were examined via western blotting and annexin V staining.

Results—Immunohistochemically, staining for 5-lipoxygenase was most frequently of greatest intensity in oral SCCs, whereas staining of cutaneous and UV radiation–induced lesions had less consistent 5-lipoxygenase expression. Exposure of SCCF1 cells to the 5-lipoxygenase inhibitor tepoxalin resulted in apoptosis; the effect appeared to be mediated via alteration of cell signaling rather than via suppression of lipid mediators that are typically produced as a result of 5-lipoxygenase activity.

Conclusions and Clinical Relevance—In cats, expression of 5-lipoxygenase in SCCs appeared to differ depending on tumor location. The influence of tepoxalin-induced 5-lipoxygenase inhibition on a 5-lipoxygenase–expressing cell line coupled with the notable expression of 5-lipoxygenase in oral SCCs suggested that 5-lipoxygenase inhibition may have therapeutic benefits in affected cats. Although the safety of tepoxalin in cats has yet to be investigated, 5-lipoxygenase inhibitors should be evaluated for use as a potential treatment for SCCs in that species.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To determine proinflammatory gene expression, endothelial adhesion molecule gene expression, and matrix metalloproteinase (MMP) concentrations in laminar specimens at 1.5 hours after administration of black walnut extract (BWE) and to compare these values with later time points.

Animals—25 horses.

Procedures—After nasogastric administration of BWE, anesthesia was induced at 1.5 hours in early time point (ETP) horses (n = 5), between 3 and 4 hours in developmental time point horses (5), and between 9 and 10 hours in acute onset of lameness time point horses (5). Anesthesia was induced at 3 and 10 hours after nasogastric administration of water in 2 groups of control horses (3-hour control group, n = 5; 10-hour control group, 5). Real-time quantitative PCR assay was performed on laminar specimens from control and ETP horses for cyclooxygenase (COX)-1, COX-2, interleukin (IL)-1β, tumor necrosis factor-α, IL-6, IL-8, IL-10, MMP-2, and MMP-9 gene expression; and on laminar specimens from all groups for endothelial adhesion molecules, intercellular adhesion molecule (ICAM)-1, and E-selectin gene expression. Leukocyte emigration was assessed via CD13 immunohistochemistry, and gelatinase accumulation was determined by gelatin zymography.

Results—Laminar concentrations of IL-1β, IL-6, IL-8, COX-2, ICAM-1, and E-selectin mRNA were significantly increased in ETP horses, compared with control horses. Concentrations of IL-1β, IL-8, ICAM-1, and E-selectin mRNA peaked at 1.5 hours. In ETP horses, leukocyte emigration was present in 3 of 5 horses and pro–MMP-9 was detected in 2 of 5 horses.

Conclusions and Clinical Relevance—Results indicated that endothelial activation and laminar inflammation are early events in laminitis; MMP accumulation likely is a downstream event.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To determine the difference in histologic artifacts and morphologic diagnosis among 3 laparoscopic cup biopsy forceps techniques and wedge hepatic samples.

ANIMALS

Cadavers of 20 client-owned dogs following euthanasia for unrelated reasons between January 3 and July 29, 2021.

PROCEDURES

Four biopsy techniques were performed from the margin of 3 liver lobes/dog. Laparoscopic techniques used 5-mm cup biopsy forceps to obtain biopsy samples by pulling the forceps forcefully caudally to free a sample (the PULL technique), rotating the forceps 360° in 1 direction until freed (the TWST technique), or pulling the forceps through a 5-mm cannula to remove the sample (the CAN technique); wedge biopsy samples served as the control (CON). Data collected included sample weight, histologic features, diagnosis, and artifact characterization. Gwet AC1 or intraclass correlation coefficients (ICCs) were calculated to detect agreement among techniques.

RESULTS

Sample weights for CON and TWST were significantly larger (P < .001 and P = .035, respectively) than for PULL and CAN. There was excellent agreement among all techniques for most diagnostic features (Gwet AC1, 0.93 to 1). The TWST technique resulted in the best overall artifact profile for laparoscopic techniques, with 90% of samples (54/60) having crisp edges and 65% of samples (39/60) having no or mild tearing. The agreement was moderate to good (ICC, 0.73 for edges and 0.76 for tearing) among all cup biopsy forceps techniques.

CLINICAL RELEVANCE

The TWST technique resulted in the largest sample and had the fewest artifacts, supporting its continued use during laparoscopic procedures.

Open access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To characterize aminoaciduria and plasma amino acid concentrations in dogs with hepatocutaneous syndrome (HCS).

ANIMALS 20 client-owned dogs of various breeds and ages.

PROCEDURES HCS was definitively diagnosed on the basis of liver biopsy specimens (n = 12), gross and histologic appearance of skin lesions (4), and examination of skin and liver biopsy specimens (2) and presumptively diagnosed on the basis of cutaneous lesions with compatible clinicopathologic and hepatic ultrasonographic (honeycomb or Swiss cheese pattern) findings (2). Amino acid concentrations in heparinized plasma and urine (samples obtained within 8 hours of each other) were measured by use of ion exchange chromatography. Urine creatinine concentration was used to normalize urine amino acid concentrations. Plasma amino acid values were compared relative to mean reference values; urine-corrected amino acid values were compared relative to maximal reference values.

RESULTS All dogs had generalized hypoaminoacidemia, with numerous amino acid concentrations < 50% of mean reference values. The most consistent and severe abnormalities involved glutamine, proline, cysteine, and hydroxyproline, and all dogs had marked lysinuria. Urine amino acids exceeding maximum reference values (value > 1.0) included lysine, 1-methylhistidine, and proline.

CONCLUSIONS AND CLINICAL RELEVANCE Hypoaminoacidemia in dogs with HCS prominently involved amino acids associated with the urea cycle and synthesis of glutathione and collagen. Marked lysinuria and prolinuria implicated dysfunction of specific amino acid transporters and wasting of amino acids essential for collagen synthesis. These findings may provide a means for tailoring nutritional support and for facilitating HCS diagnosis.

Full access
in American Journal of Veterinary Research