Direct effect of hypothalamic neuropeptides on the release of catecholamines by adrenal medulla in sheep - study ex vivo.

Stress causes the activation of both the hypothalamic-pituitary-adrenocortical axis and sympatho-adrenal system, thus leading to the release from the adrenal medulla of catecholamines: adrenaline and, to a lesser degree, noradrenaline. It has been established that in addition to catecholamines, the adrenomedullary cells produce a variety of neuropeptides, including corticoliberine (CRH), vasopressin (AVP), oxytocin (OXY) and proopiomelanocortine (POMC) - a precursor of the adrenocorticotropic hormone (ACTH). The aim of this study was to investigate adrenal medulla activity in vitro depending, on a dose of CRH, AVP and OXY on adrenaline and noradrenaline release. Pieces of sheep adrenal medulla tissue (about 50 mg) were put on 24-well plates and were incubated in 1 mL of Eagle medium without hormone (control) or supplemented only once with CRH, AVP and OXY in three doses (10-7, 10-8 and 10-9 M) in a volume of 10 µL. The results showed that CRH stimulates adrenaline and noradrenaline release from the adrenal medulla tissue. The stimulating influence of AVP on adrenaline release was visible after the application of the two lower doses of this neuropeptide; however, AVP reduced noradrenaline release from the adrenal medulla tissue. A strong, inhibitory OXY effect on catecholamine release was observed, regardless of the dose of this hormone. Our results indicate the important role of OXY in the inhibition of adrenal gland activity and thus a better adaptation to stress on the adrenal gland level.

Thyroid gland in vitro activity and reactivity to TRH in newborn lambs.

The aim of the study was to determine the thyroid gland tissue concentration and activity on T4 and T3 in vitro release to the medium without (control) or with TRH (10 µg/ml) for 90 min of incubation in lambs just after birth and after the first 6h of postnatal life. The results obtained show that T4 concentration in the blood plasma in newborn lambs as well after 6h of life is higher than in thyroid gland tissue. In the case of T3 the concentration in blood plasma of lambs just after birth is lower than after 6h of postnatal life. Inversely, the higher level of that hormone concentration in blood plasma just after birth is higher than in the thyroid gland tissue. The results clearly indicate the inhibitory TRH effect on in vitro T4 especially, and in lesser degree T3 release from the thyroid gland tissue of lambs just after birth. Stimulation of the TRH thyroid gland tissue from lambs after 6h of life caused an increase in T4 release, especially after 60 min of the experiment. Similarly, a significant increase of T3 release was found only after 60 min of incubation. The transient promoting TRH effect on the amount of iodothyronine release from the thyroid gland in older lambs is probably connected with this gland's function not occurring before the designated time of postnatal adaptation. The described in vitro experiment visibly shows that changing thyroid gland activity in newborn lambs is not only conditioned by the entire HPT activity, but by the immanent thyroid gland attributes as well.

Verapamil - L type voltage gated calcium channel inhibitor diminishes aggressive behavior in male Siamese fighting fish.

Verapamil is a L-type voltage gated calcium channels inhibitor (VGCCI), which is a highly prescribed drug used in the treatment of hypertension, angina pectoris, cardiac arrhythmia and cluster headaches. Its common use caused its appearance in water environment. VGCC inhibit epinephrine release and cause many neuro-hormonal changes influencing also fish behavior. Siamese fighting fish was chosen to study the influence of verapamil given to the water on the beginning of experiment in 3 different concentrations of 0 (control), 8 and 160 µg · L-1, on aggressive behavior in these fish. The experimental fish were placed in individual glass containers for 3 weeks and the mirror test was used. The highest concentration led to a significant modulation of fish behavior after 1 week and the lower dose caused statistically significant behavioral changes after 2 weeks of verapamil treatment. Siamese fighting fish males exposed to verapamil had longer latencies to the first chase - 12.6 s (8 µg · L-1 of verapamil) and 18.8 s (160 µg · L-1 of verapamil) compared to 5.6 s in the control group, decreased attack frequency and shorter duration of these attacks. The number of attacks within 10 min was decreased from 38.3 in the control group to 27.1 and 16.1, respectively. Also the total duration of these attacks decreased from 354.8 (control) to 326.4 (decrease statistically insignificant) and to 194.8 s in verapamil treated groups. It was shown, that even relatively low concentrations of verapamil in water may have adverse effects on fish and probably other living organisms.

Vaccinia virus-induced changes in cytokine-regulated acute phase plasma protein synthesis by hepatoma cells.

Human hepatoma cell line, HepG2, has been infected with vaccinia virus and synthesis of plasma proteins was determined by electroimmunoassay and corresponding mRNA's measured by Northern blotting. The inhibitory effect of the virus was dose- and time-dependent. Electrophoretic mobility shift assay revealed a decrease in C/EBP binding activities in nuclear extracts isolated from the infected hepatoma cells. Supershift analysis of the C/EBP isoforms showed alpha and beta subunit involvement in DNA binding. The treatment of the cells with interleukin-1, interleukin-6, and dexamethasone at the initial stage of infection appears to delay the virally induced inhibition of host cell protein synthesis. Thus, possible "protective" role of the acute phase cytokines in viral infection is proposed.

Induction of DNA synthesis by microtubule depolymerization is mediated by actin filaments.

Colcemid (0.4 microgram/ml) induced DNA synthesis in quiescent chick embryo fibroblasts, and increased the amount of F-actin. Cytochalasin D, which disorganized actin filaments, inhibited the stimulation of DNA synthesis by colcemid. These results suggest that actin filaments are involved in the sequence of events leading from the depolymerization of microtubules to the initiation of DNA synthesis. They also demonstrate that depolymerization of microtubules can influence the amount of F-actin in the cell.

[Effect of phenothiazine derivative on adrenal cortex response of sheep to repeated emotional stress]. / Wplyw pochodnej fenotiazyny na reakcje kory nadnerczy owiec w czasie wielokrotnego stresu emocjonalnego.

The study was aimed at the evaluation of propiopromazine (Combelen, Bayer), a derivative of phenothiazine, as an agent lowering in sheep the response to stress. The stress of emotional origin was induced in sheep by the isolation from herd lasting 1 hour. The isolation experiments were repeated 6 times on the same group of sheep, first three isolations (1-3) in daily intervals and next three (4-6) in weekly intervals. Propiopromazine was administered before each isolation experiment. The reaction of sheep to the isolation stress was weaker after propiopromazine administration. This was suggested by smaller increase in blood serum cortisol and glucose levels when compared to sheep subjected to isolation but not receiving the drug. Such effect was especially conspicuous during the course of the first isolation experiment; during the next experiments the difference concerning the reaction to stress between the sheep isolated from the herd receiving and not receiving the drug was gradually diminishing. It was shown in addition that propiopromazine administration to the sheep not subjected to stress caused an increase in cortisol level by 125 per cent and that in glucose level by 35 per cent. These results suggest that propiopromazine administration protects the organism against the effects of emotional stress only partially. Moreover, the effect of its administration gradually weakens with repeating of the stress inducing experiment, and propiopromazine itself may act as a stress inducing factor. It seems therefore that the use of propiopromazine and similar compounds as anti-stress agents may be questionable.

Food deprivation suppresses stress-induced rise in catabolic hormones with a concomitant tendency to potentiate the increment of blood glucose.

The organism of a food-deprived animal is directed toward minimizing energy expenditure and plasma levels of catabolic hormones and glucose are also reduced. Stress, on the other hand, is associated with enhancement of metabolic processes, elevated plasma catabolic hormones, and higher glucose levels. The question arises as to whether food deprivation may be able to attenuate the rise of plasma catabolic hormones seen in stress. For this purpose the variations in triiodothyronine (T3), thyroxine (T4), cortisol and glucose in blood plasma of sheep were monitored during 101 hr of food deprivation and 5 hr of stress. Stress was evoked by isolation of individual sheep from the flock. Blood was sampled by venipuncture once a day during 4 days preceding the isolation stress. On the day of isolation, blood was taken 4 times at 1.5- to 2-hr intervals. Food deprivation lowered the T3, T4 and glucose levels to 45.0, 59.5 and 78.0 percent of the basal level, respectively. Plasma cortisol level did not change over the fasting period in sheep not having visual contact with fed animals. Maintaining such a contact elevated cortisol level maximally by 139 percent over basal level. This indicates that the involvement of an emotional factor seems to be necessary for manifestation of stress. Isolation stress acting on fed and fasting sheep increased all measured hormones and glucose levels. However, in fed sheep, the maximal levels of T3, T4 and cortisol were 72.5, 48.4 and 50.0 percent higher than in corresponding isolated and food-deprived animals. Inversely, the maximal concentration of plasma glucose was about 16.6 percent higher in food-deprived than in fed animals.(ABSTRACT TRUNCATED AT 250 WORDS)