Preliminary transcriptomic analysis of peripheral blood from German Shepherd dogs with degenerative joint disease for the identification of diagnostic biomarkers.

OBJECTIVE: Degenerative joint disease (DJD) includes a group of disorders characterised by the deterioration of the articular cartilage. In this study, we investigated the transcriptomic profile of peripheral blood in German Shepherd dogs with DJD to identify putative diagnostic biomarkers. METHODS: Differential gene expression (DGE) and gene ontology (GO) analyses of the bulk RNA-seq experiment were performed in a cohort of 12 adult dogs (five cases and seven controls, classified by clinical and radiographic analyses). RESULTS: Radiographs of cases revealed severe signs of progressive DJD. Two up-regulated (LOC106559672 and THBS4) and one down-regulated (LOC106559235) differentially expressed genes (adjusted p value < 0.05) were identified. The DGE with log2 fold change < -1.5 and > 1.5 and non-adjusted p < 0.01 were selected for GO analysis. No significant enrichment terms were observed in the selected threshold. CONCLUSION: The gene-encoding protein THBS4 is correlated with DJD severity and long noncoding RNA LOC106559235 is probably involved in the DJD process. The THBS4 gene should be considered a good biomarker for DJD in dogs. Future studies using independent cohorts will be necessary to validate the present results.

Ovarian and uterine two-dimensional and Doppler ultrasonographic changes in the feline peri-estrus period.

OBJECTIVES: The aim of this study was to evaluate two-dimensional and Doppler ultrasonographic changes of the ovary and uterus during estrus and the early post-estrus period in domestic cats. METHODS: Two-dimensional and Doppler ultrasonographic evaluations of the ovaries and uterus were performed in seven queens on days 1, 3 and 5 of estrus, and 5 days after estrus (AE5). RESULTS: On day 1, 5.4 ± 0.5 follicles of 2.1 ± 0.1 mm were detected progressively increasing in number and size up to day 5 and then decreased on AE5 (P <0.05). A maximum follicular diameter of 4.1 ± 0.1 mm was achieved on day 5. Both during and after estrus, the uterus was generally hypoechoic compared with the surrounding tissues and delineated by a thin hyperechoic line corresponding to the serosa. In some animals, the uterine layers were distinguished during and after estrus. The blood flow waveforms of the intraovarian and uterine arteries were characterized by a systolic peak and diastolic flow extending throughout the remainder of the cardiac cycle to the next systole. In the uterine artery waveforms, the early diastolic notch was mild or absent during most estrous observations. The resistance index of the intraovarian arteries decreased up to day 5, and then increased on AE5 (P <0.05). The resistance index of the uterine arteries was lower during estrus than in the post-estrus period (P <0.05). CONCLUSIONS AND RELEVANCE: It is concluded that in the domestic cat, follicular number and diameter as well as ovarian and uterine blood flow changed during and immediately after estrus. Doppler ultrasound proved suitable to evaluate the hemodynamic changes involved in the cyclic remodeling of ovarian and uterine tissues that occurs during and after follicular growth in domestic cats.

Ultrasonographic evaluation of ovarian morphology and blood flow in prepubertal and pubertal cats.

The aim of this study was to evaluate two-dimensional and Doppler ultrasonographic changes of feline ovaries before and during puberty. Nine, 3-month-old female cats were followed until puberty (Day 1). Two-dimensional and Doppler ultrasound evaluations of the ovaries were carried out on Days -50, -20, -7 and 1. Longitudinal and transverse sections of the ovaries were measured and all anechoic spherical structures were considered to be follicles. The number of follicles >1 mm and the maximum diameter of the largest follicle were recorded. Peak systolic velocity and end diastolic velocity of intraovarian arteries were also measured to automatically calculate the resistive index (RI). The mean ovarian longitudinal diameter increased gradually throughout the study from 8.6 to 10.7 mm (p < 0.05). While four cats presented multiple anechoic spherical structures <1 mm diameter throughout the study, the remaining five animals had these structures only on Days -50 and -20. On Days -20, -7 and 1, the mean number of follicles were 1.4 ± 0.7, 2.5 ± 0.8 and 4.8 ± 1 respectively (p < 0.01). The largest follicles at the same time points were 1.1 ± 0.2 mm, 1.9 ± 0.3 mm and 2.6 ± 0.5 mm respectively (p < 0.05). The RI of the intraovarian arteries declined throughout the study period (p < 0.01). It is concluded that, in female cats, ovarian dimensions, follicle number and intraovarian blood flow increased from 3 months of age to puberty.

Disrupting effect of androgens in postnatal female domestic cats.

To test the hypothesis that in domestic cats, postnatal androgens induce sterility, the aims of this study were to describe the reproductive effects and the clinical safety of a postnatal administration of a long term release androgen in this species. Thirteen newborn littermate female kittens were randomly assigned to one of the following treatment groups within the first 24h of birth: testosterone enanthate 12.5mg sc (TE; n=8) or Placebo (PL; n=5). The animals were subsequently assessed for fecal sexual hormones until puberty was attained and subsequently when matings occurred. After 21 days, ovulation and gestation were diagnosed. All queens were subsequently ovario-hysterectomized. Fecal testosterone concentrations differed between the treatment groups throughout the study period (P<0.05) being greater during the first 2 postnatal weeks in those of the TE group (P<0.01). Fecal estradiol was not affected by treatment (P>0.1). While all the females were receptive during the pubertal estrus (P>0.1), two TE (2/8) compared with all (5/5) females of the PL group had ovulations (P<0.05). Only one (1/2) compared with three (3/5) of the queens of the TE and PL groups, respectively became pregnant. All kittens of the TE group had transient clitoral enlargement. Anovulatory TE-treated cats had no corpus luteum, and a significant diminution of the endometrial glands as well as of the height of the uterine epithelium. It is concluded that, in domestic cats, a single postnatal supra-physiological dose of testosterone caused a large proportion of queens to be anovulatory and there were also histological endometrial abnormalities that also occurred with this treatment that were accompanied by mild and transient side effects.