In vivo evidence that local cortisol production increases in the preovulatory follicle of the cow

The aim of the present in vivo study was to monitor real-time fluctuations of cortisol (Cr) in the wall of preovulatory follicles using a microdialysis system (MDS) implanted in the theca layer as well as changes in ovarian venous plasma (OVP) and jugular venous plasma (JVP). Seven cows were superovulated using FSH and prostaglandin F2alpha injections. Dialysis capillary membranes were surgically implanted into the theca layer of mature follicles and connected to a microdialysis system. Fractions of the perfusates were collected from Day -1 (Day 0=LH surge) to Day 3. No difference in the concentrations of Cr between JVP and OVP was detected throughout the experiment. Circulating concentrations of Cr ranged from 20 to 35 ng/ml 8 h after surgery in ovulatory and anovulatory cows. In five ovulatory cows, the Cr concentration decreased to basal levels (<10 ng/ml) between 12 and 24 h after surgery, however, two anovulatory cows retained high Cr levels (>10 ng/ml) up to 42 h after surgery. There was a clear increase in the local concentration of Cr from 13.3+/-2.1 pg/ml at -24 h to 27.5+/-1.7 pg/ml at 0 h (peak of the LH surge) within the wall of ovulatory follicles. This increase was not detected in anovulatory follicles. This transient increase in Cr occurred only in the follicle wall, but not in the OVP or JVP, indicating that the presence of a local regulatory mechanism for Cr production/conversion in ovulatory follicles, and this mechanism may modulate the inflammatory-like reaction induced by LH surge in the follicle wall. The present results demonstrate that the glucocorticoid environment in the follicular wall adjusts at the local level in bovine ovulatory follicles. This mechanism may protect follicles from the adverse effects of glucocorticoid, and it may prevent excess inflammatory reactions associated with ovulation by temporarily increasing local concentrations of glucocorticoid, thus forming an integral part of the regulatory mechanism in ovarian physiology.     

Intrafollicular paraoxonase 1 activity and the steroidogenic potential of the first post‐partum dominant follicle in dairy cows

Cows experiencing high levels of inflammation and specific metabolic conditions tend to have slower follicular growth and lower serum and follicular concentrations of oestradiol (E2). Paraoxonase1 (PON1) activity decreases during inflammatory processes. Therefore, the aim of this study was to evaluate the association between serum and intrafollicular (FF) PON1 activity and the serum and intrafollicular levels of E2 and progesterone (P4), as well as the mRNA expression of genes related to steroidogenesis, metabolism and inflammation in the first post‐partum dominant follicle of Holstein cows. No correlation was found between PON1 activity, the expression of the analysed genes and levels of follicular E2 and P4, except for a negative correlation between serum E2 and follicular PO1 activity. Also, no correlation was found between serum and follicular PON1 during the first post‐partum follicular wave.     

Autoantigenic nuclear proteins of a clinically atypical renal vasculitis

Background

Systemic vasculitides constitute a heterogeneous group of diseases of autoimmunological origin characterized by inflammation of blood vessels and antibodies that react against autoantigens in a process that ultimately affects blood vessel walls. An important number of these patients present kidney disease. An endeavour of this area of research is the identification of autoantigens involved in these diseases. Accordingly, we used serum from a patient suffering from a microscopic polyangiitis, P-ANCA positive, manifesting a clinically atypical renal necrotizing glomerulonephritis and interstitial nephropathy for the identification of autoantigens; we also determined the prevalence of corresponding autoantibodies in other vasculitides, diabetic microangiopathy and in general population.

Methods

The patient's serum was used as a probe for the immunoscreening method SEREX to screen a human brain cDNA expression library.

Results

Four positive clones were isolated and sequenced. Clones Jos002 code for protein HDAC5, Jos014 for TFC4, Jos107 for RTF1, and Jos313 for POLDIP3 polymerase. The four proteins are of nuclear localization. None of them had been reported as autoantigen. Recombinant proteins were synthesised and checked as antigens by western blot with different sera from controls and patients affected with other vasculitides and diabetic microangiopathy as well. Only the serum from the patient origin of this study recognized all recombinant proteins.

Conclusion

We identify four nuclear proteins, HDAC5, TFC4, RTF1 and POLDIP3 polymerase as new autoantigens that could be used as markers in the diagnosis of subfamilies in immune diseases, although we cannot determine the role of these proteins in the aetiopathogenic process.

     

Hafnia alvei as an opportunistic pathogen causing mortality in brown trout, Salmo trutta L.

Strains of Hafnia alvei caused mortalities in brown trout, Salmo trutta L., following intraperitoneal injection with LD₅₀ values ranging between 1.3 × 10⁴ and 2.5 × 10⁷ bacteria fish^??1. These values are considered to represent a high to moderate degree of virulence. Virulent strains were isolated from non-fish sources. Fish surviving the LD₅₀ values continued to harbour the organism in the kidney, suggesting the establishment of a carrier state.     

Factors influencing half‐pearl (mabe) production and quality in the red abalone Haliotis rufescens

The factors influencing half‐pearl (mabe) production in the red abalone Haliotis rufescens were investigated, as a strategy to optimize the technology for the development of a pearling industry in Chile. The effects of abalone size (small, large), implant position (dorsal, central, ventral and their combinations), and number of nuclei (one, two, three) were analysed on the quantity, thickness of the nacre layer and quality (shape, colour, lustre, defects) of the mabe produced. The size of abalone did not exert a clear influence on the quantity and quality of mabe, but the position on the shell (firstly) and number of nuclei (secondly) did. More mabe, with thicker nacre, brighter lustre and fewer defects occurred in small or large abalone implanted with only one nucleus on dorsal position. In contrast, small and large abalone implanted in the ventral position formed less mabe, with thinner nacre, poor lustre and more defects. This likely occurs because mantle tissue folds at the ventral position and does not always cover the nucleus, leaving it partially or totally exposed. Most of the pearls were categories AA in small abalone and A in large abalone; only three AAA gems were produced in large abalone with one nucleus on dorsal position or with three nuclei (DCV). Based on these results, we recommend implanting abalone at 60–100 mm shell length, with only one nucleus on the dorsal position.     

Cryopreservation of sperm in annual fish Austrolebias minuano (Cyprinodontiformes; Rivulidae)

The effects of the cryoprotectants dimethyl sulfoxide, glycerol and methyl glycol at different concentrations on Austrolebias minuano sperm quality parameters were evaluated in this study. The cellular kinetic parameters, determined using flow cytometry, indicated the best results with the samples cryopreserved with 7.5% methyl glycol. Dimethyl sulfoxide concentrations of 7.5% and glycerol concentrations of 10%, 12.5% and 15% demonstrated the least benefit across all evaluated sperm kinetic parameters. When assessed using flow cytometry, a concentration of 7.5% methyl glycol similarly showed better sperm kinetic parameters than 12.5% dimethyl sulfoxide. We conclude that cryoprotectants, especially methyl glycol, are effective for the preservation of sperm quality in A. minuano. However, high sensitivity of spermatozoa against glycerol was observed in these studies; thus, it is not recommended for cryopreservation purposes.