The therapeutic value of protein (de)nitrosylation in experimental septic shock

Cardiovascular dysfunction and organ damage are hallmarks of sepsis and septic shock. Protein S-nitrosylation by nitric oxide has been described as an important modifier of protein function. We studied whether protein nitrosylation/denitrosylation would impact positively in hemodynamic parameters of septic rats. Polymicrobial sepsis was induced by cecal ligation and puncture. Female Wistar rats were treated with increasing doses of DTNB [5,5′-dithio-bis-(2-nitrobenzoic acid)] 30 min before or 4 or 12 h after sepsis induction. Twenty-four hours after surgery the following data was obtained: aorta response to phenylephrine, mean arterial pressure, vascular reactivity to phenylephrine, biochemical markers of organ damage, survival and aorta protein nitrosylation profile. Sepsis substantially decreases blood pressure and the response of aorta rings and of blood pressure to phenylephrine, as well as increased plasma levels of organ damage markers, mortality of 60% and S-nitrosylation of aorta proteins increased during sepsis. Treatment with DTNB 12 h after septic shock induction reversed the loss of response of aorta rings and blood pressure to vasoconstrictors, reduced organ damage and protein nitrosylation and increased survival to 80%. Increases in protein S-nitrosylation are related to cardiovascular dysfunction and multiple organ injury during sepsis. Treatment of rats with DTNB reduced the excessive protein S-nitrosylation, including that in calcium-dependent potassium channels (BK_Ca), reversed the cardiovascular dysfunction, improved markers of organ dysfunction and glycemic profile and substantially reduced mortality. Since all these beneficial consequences were attained even if DTNB was administered after septic shock onset, protein (de)nitrosylation may be a suitable target for sepsis treatment.     

Adrenocorticotropic hormone (ACTH) in pituitary colloid and extrapituitary tissues

The level of ACTH in bovine pituitary intraglandular colloid (IGC/ACTH), the holocrine secretion of the marginal half of the intermediate lobe (IL) cells, was found to correlate with the concentration of this peptide in blood plasma (BP/ACTH), cerebrospinal fluid (CSF/ACTH), the paraventricular nuclei (PVN/ACTH) and the arcuate nuclei (AN/ACTH). The ACTH content in all sites was measured by radioimmunoassay (RIA). Although the IL is virtually avascular, the intraglandular lumen housing the IGC communicates nonetheless with the venous cavernous sinuses as well as the subarachnoid cerebral spinal fluid (CSF). Therefore, we content that the IGC serves as a transport medium whereby IL materials gain access to these extrapituitary sites.     

Ultrasonography and hormone profiles of adrenocorticotrophic hormone (ACTH)-induced persistent ovarian follicles (cysts) in cattle

The objective of this study was to develop a model for the study of abnormal ovarian follicles in cattle by treating heifers with adrenocorticotrophic hormone (ACTH) (100 iu at 12 h intervals for 7 days, beginning on day 15 of the oestrous cycle). Cortisol concentrations increased (P < 0.05) within 24 h after beginning ACTH treatment and cortisol and progesterone concentrations remained elevated after cessation of ACTH treatment for 8 and 4 days, respectively. The pulses and surges of LH decreased during ACTH treatment, but FSH profiles were similar to those in controls and persistent or prolonged follicles were eventually observed in all heifers. In five heifers, prolonged dominant follicles ovulated after 10 days, whereas in six heifers, persistent follicular structures were present for 20 days, but ceased to secrete oestradiol after approximately 12 days. In the heifers with persistent follicular structures, new follicles emerged when the persistent follicle became non-oestrogenic. During the last 2 days of normal follicular growth, the concentration of oestradiol was greater than it was during prolonged or persistent follicle development (P < 0.05). There were no differences in the growth rates or maximum diameters of abnormal follicles that had different outcomes, but oestradiol concentrations were greater in prolonged follicles that ovulated compared with those follicles that persisted (P = 0.06). In conclusion, stimulation with ACTH resulted in a marked deviance from normal follicular activity. The aberrations were probably caused by the interruption of pulsatile secretion of LH (but not FSH) leading to decreased but prolonged oestradiol secretion.     

Effect of different concentrations of serotonin, histamine and insulin on the hormone (serotonin and ACTH) production of Tetrahymena in nutrient-free physiological milieu

Cell populations of Tetrahymena pyriformisGL were kept in nutrient-free (Losina) milieu and treated with different (10⁻⁶–10⁻²¹ M) concentrations of serotonin, histamine or insulin for 30 min. Following that the hormone (serotonin and adrenocorticotropin (ACTH) content of the cells were measured by immunocytochemical flow cytometric method. Serotonin reduced histamine when applied in 10⁻¹² and 10⁻¹⁵ M concentrations, while elevated ACTH levels when applied in 10⁻⁶, 10⁻⁹ and 10⁻²¹ M concentrations. Histamine reduced serotonin concentration at 10⁻⁹–10⁻²¹ M concentrations and increased ACTH in 10⁻⁶ M. Insulin elevated both hormones’ content in each concentration except at 10⁻¹² M. The results demonstrate that (1) in nutrient-free conditions the hormonal effects differ from that of nutrient-rich (tryptone + yeast) condition; (2) there is an optimal hormone concentration, which causes the strongest effect and this is different for each hormones; (3) the hormone receptors of Tetrahymena are very sensitive; as they react to zeptomolar concentrations. Such small concentration is even more effective than higher ones. Since hormones must become highly diluted in the natural environment of Tetrahymena, it seems that such low concentrations are the actual physiological concentrations.     

Secretion patterns of luteinizing hormone in growing mithuns (Bos frontalis)

To characterize the luteinizing hormone (LH) secretion patterns in growing mithun (Bos frontalis), a semi-wild ruminant, six female mithuns (1 year old; BW: 145.5 kg) were maintained in a semi-intensive system. Plasma progesterone (P(4)) level was measured in twice-a-week samples collected for six weeks to assess ovarian status. This was followed by a frequent sampling period. Blood samples collected at 15 min intervals for 9 h were assayed for plasma LH. Luteinizing hormone patterns consisted of pulses of varying amplitudes. Luteinizing hormone pulses occurred at an average rate of 0.54/h ( approximately 5 pulses/9 h). The rate did not differ among mithuns. The mean plasma LH levels was correlated with body weight (r=0.82; p<0.05) and pulse amplitude (r=0.87; p<0.01). Neither the LH amplitude nor the frequency was affected by time (p>0.05). The mean plasma P(4) concentration was 0.37 ng/ml. In conclusion, we demonstrated a pulsatile nature of LH secretion in growing mithuns. In addition, the mean plasma LH level and LH amplitude were positively correlated with body weight. It appears that in contrast to cattle, five LH pulses per nine hours recorded in mithuns were not an indication of approaching puberty.