Effects of orexin-monoaminergic interactions on oxytocin secretion in rat neurohypophyseal cell cultures

The effects of orexin-monoaminergic compound interactions on oxytocin release were studied in 14-day rat neurohypophyseal cell cultures prepared by an enzymatic dissociation technique. The oxytocin contents of the supernatants were determined by radioimmunoassay. Following the administration of orexin-A or orexin-B in increasing doses, significant changes were not observed in the oxytocin content of the supernatant media. The oxytocin level increased substantially in response to adrenaline, noradrenaline, serotonin, histamine, dopamine or K(+) treatment. Preincubation with orexin-A or orexin-B reduced the adrenaline-, histamine- or serotonin-induced oxytocin level increases, but the oxytocin concentrations of the supernatant media remained above the control level. There was no significant difference in decreasing effect between orexin-A and orexin-B. Neither orexin-A nor orexin-B induced changes in oxytocin release following monoaminergic compound treatment. The results indicate that the changes in oxytocin secretion induced by the monoaminergic system can be directly influenced by the orexin system. The effects of orexin on oxytocin release can be antagonized by an orexin-1 receptor-specific antagonist. It may be presumed that the orexins can play a role in the pathogenetic process of metabolic diseases (e.g. obesity) by reducing the effects of increased oxytocin release caused by monoaminergic compounds. The interactions between the monoaminergic and orexin systems regarding oxytocin secretion occur at both the hypothalamic and the neurohypophyseal levels.     

Major and trace elements in whole blood of phlebotomized patients with porphyria cutanea tarda

Porphyria cutanea tarda (PCT) is a disorder of hem biosynthesis resulting from a decreased activity of the uroporphyrinogen decarboxylase enzyme. Hem precursors are accumulated in the blood, liver and skin. Inherited and acquired factors also contribute to the pathogenesis of PCT. Hem precursors and porphyrins are excreted with urine and faeces. Whole blood of 8 PCT patients and 6 volunteers of Caucasian origin were analysed. In addition to routine laboratory measurements, 19 elements (Al, B, Ba, Ca, Cd, Co, Cu, Fe, K, Li, Mg, Mn, Mo, Na, Ni, P, S, V, Zn) were determined by means of inductively coupled plasma optical emission spectrometry (ICP-OES). Mg, P and S concentrations in whole blood were decreased significantly (p<0.05), whereas Ba was increased in PCT patients compared to controls. Metabolic alterations are reflected in the correlation of parameters. Positive correlations were found between the element pairs of Zn-Al, Zn-Mg, Zn-Mn, B-S, Fe-Mg, K-P, Mg-Mn for PCT patients, whereas in the control group Al-Mn, Ca-Cu, Ca-Na, Cu-Mg, Fe-K, Mg-Na, Zn-P showed positive correlations.     

A non-invasive device to continuously determine heat strain in humans

Heat stroke remains a very dangerous, potentially lethal illness in humans. The Physiological Strain Index (PSI), originally based on heart rate and rectal temperature recordings in humans, describes heat strain in quantitative terms. The objective of our study was to establish whether the rectal temperature recordings serving to determine the PSI could be replaced by a non-invasive skin temperature sensor combined with a heat flux sensor (Double Sensor) attached to the inside of a helmet. We assumed (i) that the difference between the recordings by the device under test and the rectal temperature should be less than ±1.0 °C for ±2 S.D. at 10, 25, and 40 °C ambient temperature, and (ii) that the temperature predictions based on the Double Sensor temperature should differ by less than 1 PSI score from the calculations based on recordings of the rectal temperature. Twenty male subjects participated in the study. Rectal, nasopharyngeal, and skin temperatures, heat flux, and cardiovascular data were collected continuously during different experimental setups at ambient temperatures of 10, 25, and 40 °C. Depending on the protocols, the exercise intensities varied from 25% to 55% of the individual VO_2max. A comparison of the recordings obtained from the device under test with those of the rectal temperature revealed that (i) the recordings of the Double Sensor differed by −0.16 to 0.1 °C from the mean rectal temperature, (ii) the concordance correlation coefficients (CCC) during all work and rest periods rose with rising ambient temperatures (all work periods: 10 °C: 0.49; 25 °C: 0.69; 40 °C: 0.75; all rest periods: 10 °C: 0.39; 25 °C: 0.81; 40 °C: 0.74), and that (iii) the Double Sensor in the helmet showed that during all rest periods and in all ambient conditions, the temperature dropped much more quickly than what was recorded when taking the rectal temperature (p<0.01). When we compared the PSI values based on the rectal temperature recordings to those determined by the Double Sensor, it was found that (i) the PSI based on the Double Sensor recordings differed by −0.27 to 0.17 from the mean PSI established by rectal temperature recordings. Furthermore, the CCC for the PSI rose during all work periods (10 °C: 0.81, 25 °C: 0.93, 40 °C: 0.87) and rest periods (10 °C: 0.68; 25 °C: 0.93; 40 °C: 0.79). In conclusion, under warm/hot environmental conditions the device under test provided a reliable method of assessing the PSI in operational environments to improve physiological situational awareness and safety in action. However, there are some limitations that reduce the device's performance in cold environments; these need to be investigated further.     

Adenomatous polyposis coli proteins and cell adhesion

Adenomatous polyposis coli (APC) is an important tumour suppressor in the mammalian intestinal epithelium. It binds to beta-catenin and its role as a tumour suppressor depends predominantly on its ability to downregulate soluble beta-catenin, a key effector of the Wnt signalling pathway. However, epithelial cells have a distinct subcellular pool of beta-catenin, or Drosophila Armadillo, which functions as a structural component of adherens junctions. Notably, APC proteins can be associated with these adherens junctions, and recent evidence points to a role for APC in cellular adhesion. Thus, APC--like beta-catenin/Armadillo--may have a dual role in Wnt signal transduction and in cellular adhesion, which could be relevant to its activity as a tumour suppressor.     

Intermolecular relations between the glucocorticoid receptor, ZAP-70 kinase, and Hsp-90

The glucocorticoid receptor (GR) participates in both genomic and non-genomic glucocorticoid hormone (GC) actions by interacting with other cytoplasmic signalling proteins. Previously, we have shown that high dose Dexamethasone (DX) treatment of Jurkat cells causes tyrosine phosphorylation of ZAP-70 within 5 min in a GR-dependent manner. By using co-immunoprecipitation and confocal microscopy, here we demonstrate that the liganded GR physically associates with ZAP-70, in addition to its phosphorylation changes. The association of the ligand-bound GR and ZAP-70 was also observed in HeLa cells transfected with ZAP-70, suggesting that this co-clustering is independent of lymphocyte specific factors. Furthermore, the ZAP-70 was found to also co-precipitate with Hsp-90 chaperone both in Jurkat and transgenic HeLa cells, independent of the presence of DX. These findings raise the possibility that ZAP-70 may serve as an important link between GC and TcR-induced signaling, thereby transmitting non-genomic GC action in T-cells.     

Gamma irradiation induced apoptotic changes in the chromatin structure of human erythroleukemia K562 cells

Exponentially growing human erythroleukemia K562 cells were synchronized by centrifugal elutriation prior to and after Co⁶⁰ γ-irradiation (4 Gy). Forward scatter flow cytometry used for size analysis revealed the increase of an early apoptotic cell population ranging from lower (0.05 C-value) to higher DNA content (∼1 C) as the cells progressed through the S phase. The increase in cellular DNA content expressed in C-values correlated with apoptotic chromatin changes manifested as many small apoptotic bodies in early S phase and larger but less numerous disintegrated apoptotic bodies in late S phase. Most significant changes after exposure to γ-irradiation took place in early S phase resulting in an increase of nuclear size by more than 50%. Cell fractions containing irradiated cells showed enhanced growth arrest at 2.4 C-value, which was accompanied by apoptosis. Apoptotic cell cycle arrest near to the G₁/G₀ checkpoint and apoptotic changes indicate that the radiation resistance of K562 cells is related to the bypass of the early stage of the p53 apoptotic pathway. Apoptotic changes in chromatin structure induced by γ-irradiation indicate that these injury-specific changes can be identified and distinguished from chromatin changes induced by UV radiation or heavy metals.