Immunohistochemical Expression of Growth Factors in the Follicular Wall of Normal and Cystic Ovaries of Sows

The expression of growth factors was evaluated immunohistochemically in normal and cystic ovaries of sows. The immunohistochemically stained area (IHCSA) was quantified by image analysis to analyse the expression of these proteins in the follicular wall of secondary, tertiary and cystic follicles. IGF‐I immunoreactivity was strong in the granulosa cell layer (GC), moderate in the theca interna (TI) and mild in the theca externa (TE) of the normal follicles. There was severe reduction of the labelling to IGF‐I in the GC of the follicular and luteinized cysts. In the normal follicles, the reactivity for IGF‐II was very similar to pattern noted in IGF‐I. There was reduction of the IHCSAs in the GC of the follicular and luteinized cysts, but the decrease was not significant. The staining of the IGF‐II in the TI and TE of the cysts was increased, in comparison with normal follicles. The IHCSAs for VEGF were higher in the GC and TE of the normal follicles in contrast to TI, but this difference was noted only in the tertiary follicle. The VEGF reactivity increased in the GC of the cysts, in relation to normal follicles. The results of the current study show that the formation of ovarian cysts in sows is associated with alterations in the immunohistochemical expression of some growth factors.     

Unconditional quantum teleportation

Quantum teleportation of optical coherent states was demonstrated experimentally using squeezed-state entanglement. The quantum nature of the achieved teleportation was verified by the experimentally determined fidelity Fexp = 0.58 +/- 0.02, which describes the match between input and output states. A fidelity greater than 0.5 is not possible for coherent states without the use of entanglement. This is the first realization of unconditional quantum teleportation where every state entering the device is actually teleported.     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

Direct measurement of femtomoles of osmium and the 187Os/186Os ratio in seawater

Two depth profiles of the osmium concentration and the 187Os/186Os isotopic ratio in the Indian Ocean showed that the osmium concentration seems to be unaltered by chemical or biological processes occuring in seawater; accordingly, osmium is conservative. These data were obtained from an experimental method that eliminated the problems related to osmium preconcentration. This method led to a new evaluation of the concentration of osmium in seawater; the mean concentration of osmium and the 187Os/186Os ratio are equal to 10.86 +/- 0.07 picograms per kilogram and 8.80 +/- 0.07, respectively. The results suggest the existence of an organocomplex that dominates the speciation of osmium in seawater.     

Isolation of very virulent Marek''s disease viruses from vaccinated chickens in Australia

Very virulent Marek's disease viruses (vvMDV), defined as isolates against which the herpesvirus of turkey (HVT) vaccine provide poor protection, have been isolated from poultry flocks in both the United States and Europe. Twenty-one samples from vaccinated Australian flocks, experiencing problems with excessive Marek's disease (MD), were tested for the presence of transmissible MD viruses (MDV). Of the 16 samples which contained a transmissible agent, 14 were pathogenic in chickens, based on the development of MD lesions or depression of the bursa/body weight ratio. Of the pathogenic isolates which have been successfully typed 10 were serotype 1, and one was serotype 2 MDV. Pathogenicity of isolates varied. Several isolates caused tumours in 20-30% of both vaccinated and unvaccinated chickens. Two isolates, MPF6 and MPF23, caused tumours in more than 50% of chickens. When MPF6 and MPF23 were tested in vaccine trials bivalent vaccine gave no better protection against development of MD lesions than a monovalent vaccine. Isolate MPF23 was so pathogenic that lesions were produced in all chickens, regardless of the vaccine protocol used. Therefore vvMDV have been isolated in Australia, and unlike the vaccines tested overseas, bivalent Australian vaccines do not appear to provide greater protection against these vvMDV.     

Metabolic clearance rate of cortisol in pigs: relationship to adrenal responsiveness

Experiments were conducted on 12 prepuberal (18- to 20-week-old) Landrace cross Large White gilts to establish if differences in adrenal responsiveness between individuals could be explained by differences in the metabolic clearance rate (MCR) of cortisol. Pigs with the highest (n = 6) and lowest (n = 6) cortisol concentration 60 minutes after challenge with ACTH were selected from a pool of 36 commercial pigs. Tritium-labelled cortisol was infused (17 to 27 ml h-1) continuously for 120 minutes to establish 'steady state' conditions. Blood samples (10 ml) were collected at 90, 100, 110 and 120 minutes. Replicate experiments were performed on some pigs. Classification of individual pigs as high or low adrenal responders to ACTH challenge was confirmed at the end of the clearance rate experiments. The MCR of cortisol in the group classed as low adrenal responders was 59.7 +/- 7.8 litres h-1 or 1.01 litres h-1 kg-1 (n = 7) which was not significantly different from the average MCR in the group classed as high adrenal responders 60.2 +/- 5.9 litres h-1 or 1.19 litres h-1 kg-1 (n = 10). These results suggest that the repeatable differences in adrenal responsiveness to ACTH that exist between individuals within a particular strain of pig depend on differences in the rate of synthesis of cortisol in response to ACTH stimulation, rather than on differences in its rate of metabolism.     

Pituitary and adrenocortical responses to corticotropin-releasing factor in pigs

A study was conducted to determine whether differences in adrenocortical response to exogenous adrenocorticotrophin (ACTH) were an accurate reflection of an animal's perception of and response to stressful stimuli, or whether the pituitary gland might modulate adrenocortical responsiveness. Sixteen Large White x Landrace female pigs, of which 8 had high adrenocortical response to ACTH and the other 8 had low response, were administered IV a bolus of synthetic human corticotropin-releasing factor (hCRF) at dose rates ranging from 0.002 to 2 micrograms/kg of body weight. Blood samples were collected at known times for up to 2 hours after administration of hCRF. Plasma ACTH and cortisol concentrations were measured by radioimmunoassay. Results indicate that hCRF stimulated the pituitary gland of high- and low-responding pigs to secrete ACTH, which in turn stimulated the adrenal cortex to secrete cortisol. Plasma ACTH concentration, before or after hCRF administration, was not significantly different between the high and low responders. However, high-responding pigs had higher cortisol concentration after hCRF administration than did low-responding pigs. Thus, the differences in adrenocortical response to ACTH between the 2 groups of pigs were not attenuated by variation in pituitary response. It is concluded that adrenocortical responsiveness to ACTH is an accurate indicator of the perception of and the response to stress.     

Consistent capacity for adrenocortical response to ACTH administration in pigs

The effect of ACTH administration on plasma cortisol concentrations in purebred and crossbred pigs was investigated. Pigs were given either 25 IU of ACTH or physiologic saline solution IM. Blood samples were collected immediately before and 1 hour after ACTH or saline solution administration. Administration of ACTH resulted in a significant (P less than 0.01) increase in plasma cortisol concentration compared with that resulting from administration of saline solution; mean values after ACTH administration were similar in both breed groups. In contrast, a fivefold range of differences was observed among individual pigs of the same age, sex, and body weight, irrespective of breed group. The type and magnitude of the adrenocortical response was consistent and repeatable in pigs over a 3-month period, suggesting that pigs have a consistent capacity for adrenocortical response to ACTH administration. Development of a dynamic test allowed the high and low responding extremes in a population to be detected. The most suitable dose of synthetic ACTH was established to be 50 IU, and the best time for blood sample collection was 60 minutes after ACTH administration. The classification of individual pigs as high or low responders was repeatable and was not affected by prior short-term exposure to ACTH.