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  • Author or Editor: Jun Wang x
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Abstract

Objective—To evaluate the changes in concentrations of matrix metalloproteinase (MMP)-2 and MMP-9 in the precorneal tear film of dogs with Pseudomonas aeruginosa–associated keratitis during corneal healing and stromal remodeling.

Animals—10 dogs with unilateral P aeruginosa–associated keratitis and 10 clinically normal dogs.

Procedures—Precorneal tear film samples were collected from both eyes of 10 dogs with unilateral P aeruginosa–associated keratitis on the day of admission to the hospital and then at various time points until complete healing of the cornea was achieved. Precorneal tear film samples were also collected from both eyes of 10 clinically normal adult dogs (control group). Concentrations of MMP-2 and MMP-9 in precorneal tear film samples from each group were determined via gelatin zymography for comparison.

Results—The proteolytic processes in the ulcerated eyes decreased as corneal healing progressed. On the day of admission, concentrations of latent and active forms of MMP-2 and MMP-9 in ulcerated eyes were significantly higher than values in the contralateral unaffected eyes in dogs with P aeruginosa–associated keratitis; concentrations of latent MMP-2 and MMP-9 were also greater than control group values. Concentrations of latent and active forms of MMP-2 and MMP-9 in the healed eyes of dogs with P aeruginosa–associated keratitis were significantly lower than concentrations in the ulcerated eyes on the day of admission.

Conclusions and Clinical Relevance—Results suggest that reduction of precorneal tear film concentrations of MMPs by use of proteinase inhibitors may be effective in the treatment of dogs with P aeruginosa–associated keratitis.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To evaluate a hypervariable octameric oligonucleotide fingerprints (HOOF-Prints) assay for identification of and discrimination between wild-type and vaccine strains of Brucella melitensis.

SAMPLE Brucella melitensis vaccine strain M5 and wild-type strain M43.

PROCEDURES 8 pairs of primers (alterable, octameric nucleotides) were designed on the basis of a biological analysis of 8 flanking sequences in the DNA of B melitensis. The HOOF-Prints technique was used to identify wild-type and vaccine strains of B melitensis. Phylogenetic analysis of short, polymorphic fragments of DNA from B melitensis strains M5 and M43 was performed.

RESULTS Variable-number tandem repeat DNA segments of B melitensis vaccine strain M5 and wild-type strain M43 were successfully amplified by means of PCR assay. All target gene fragments ranged in size from 100 to 300 bp. Separate phylogenetic analysis of each Brucella strain revealed considerable differences between the vaccine and wild-type strains.

CONCLUSIONS AND CLINICAL RELEVANCE The results of this study suggested the HOOF-Prints assay may be useful for discriminating vaccine strains of B melitensis from wild-type strains. This ability could allow discrimination between animals that are seropositive because of vaccination against B melitensis and those that are seropositive because of B melitensis infection and could decrease the likelihood of importing Brucella-infected animals.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

Histone deacetylases (HDACs) are the key regulators involved in the process of embryo development and tumor progression and are often dysregulated in numerous disordered cells, including tumor cells and somatic cell nuclear transfer (SCNT) embryos. Psammaplin A (PsA), a natural small-molecular therapeutic agent, is a potent histone deacetylase inhibitor (HDACi) that alters the regulation of histone.

SAMPLES

Approximately 2,400 bovine parthenogenetic (PA) embryos.

PROCEDURES

To investigate the effect of PsA on bovine preimplanted embryos, we analyzed the preimplantation development of PA embryos treated with PsA in this study.

RESULTS

The blastocyst formation rate of bovine PA embryos decreased sharply with an increase in concentration and duration. Furthermore, the expression of the pluripotency-related gene Nanog was decreased, and the inhibitory effects on histone deacetylases 1 (HDAC1) and DNA methylation transferase 1 (DNMT1) were observed in bovine PA embryos. The acetylation level of histone H3 lysine 9 (H3K9) was enhanced by a PsA treatment of 10 μM for 6 h, while the DNA methylation appeared unchanged. Interestingly, we also found that PsA treatment enhanced the intracellular reactive oxygen species (ROS) generation and decreased the intracellular mitochondrial membrane potential (MMP)- and superoxide dismutase 1 (SOD1)-induced oxidative stress. Our findings improve the understanding of HDAC in embryo development and provide a theoretical basis and reproduction toxicity evaluation for the application of PsA.

CLINICAL RELEVANCE

These results indicate that PsA inhibits the development of bovine preimplantation PA embryos, supplying data for the PsA clinical application concentration to avoid reproductive toxicity. In addition, the reproduction toxic effect of PsA may be modulated through increased oxidative stress on the bovine PA embryo, suggesting that PsA in combination with antioxidants, for example, melatonin, might be an effective clinical application strategy.

Open access
in American Journal of Veterinary Research