Detection of Ultraweak Photon Emission (UPE) from Cells as a Tool for Pathological Studies.

OBJECTIVE: It is well-known that all living cells emit ultra-weak photon emission (UPE), which is due to byproducts of chemical reactions in cell metabolisms. It has been shown that Reactive Oxygen Species (ROS) in the cells enhances the UPE intensity. The magnitude of such UPE is extremely weak (i.e. a few to 103 photons/ (sec.cm22)), and the detection of such ultra-weak signals is hardly possible via sensitive instruments like photomultiplier tube (PMT) that can detect single photons. MATERIALS AND METHODS: H2O2 factor with various concentrations was applied on the HT-29 cells to generate ROS. H2O2 concentrations were so low to be nondestructive to the cells. Then, the effect of ROS generation on UPE intensity was investigated. PMT was used to detect UPE from HT-29 cells. RESULTS: The topical application of H2O2 was significantly different (P < 0.05) in comparison with HT-29 cells without H2O2 at a concentration of 1mM in 5 min detection time. The integrated UPE in the presence of H2O2 at concentration of 3mM was significantly higher (P < 0.05) than the integrated UPE in other groups at the same detection time. The difference between the concentrations of 3mM and 4mM was not significant (P > 0.01) for integrated UPE in the cell groups in the presence of H2O2. CONCLUSION: The results show that the recorded UPE from HT-29 cells increased with the topical application of exogenous ROS inducer. As a result, UPE can be used as a non-invasive technique for monitoring ROS in cells.

High levels of corticosterone, and gene expression of star, cyp17a2, hsd3b, cyp21, hsd11b2 during acute stress in common carp with interrenal hyperplasia.

We investigated the acute stress response in a common carp strain (E5) with interrenal hyperplasia due to 17α-hydroxylase deficiency, and in an isogenic standard (STD) carp strain. Cortisol, corticosterone and the head kidney-somatic index were measured during and after a 3 h net confinement stress. Star, cyp17a2, hsd3b, cyp21, hsd11b2 mRNA levels were measured in head kidneys using real-time qPCR. The results show very high corticosterone levels and enlargement of the head kidney in E5 fish. This is the first report in a teleost fish showing a significant increase of corticosterone levels in response to stress due to interrenal hyperplasia. The high levels of corticosterone in E5 suggest that corticosterone is not converted to aldosterone in common carp. star and hsd3b mRNA levels were significantly higher in E5 compared to STD fish, while cyp17a2 levels were significantly lower in E5. In contrast to E5, star levels did not change during stress and recovery in STD, suggesting that the enzyme is regulated in a different manner in E5 and STD fish. In E5, the levels of cyp17a2 dropped below control values after 20 min stress. These findings strongly suggest that cyp17a2 is impaired at (post)-transcriptional level. As a consequence the accumulated precursor (pregnenolone) is not converted to cortisol, but to corticosterone. In contrast to STD, significant levels of cortisol could not be detected in E5. Finally, hsd11b2 mRNA levels were significantly lower in E5 compared to STD, and did not change during stress and recovery. These results support the idea that hsd11b2 is involved in the conversion of physiologically active cortisol to inactive cortisone, as reported earlier for STD carp. In conclusion our results show high levels of corticosterone in E5 and differences in star and mRNA levels of steroidogenic genes between E5 and STD carp during net confinement stress.

Effects of Watercress (Nasturtium nasturtium) extract on selected immunological parameters of rainbow trout (Oncorhynchus mykiss).

Watercress (Nasturtium nasturtium) is a medical plant containing diverse chemically-active substances with biological properties. The present study was conducted to investigate the immunomodulatory effects of watercress extract on immunological and hematological parameters of rainbow trout (Oncorhynchus mykiss). Fish were fed for 21 days with diet supplemented with 0.1% and 1% of watercress extract per 1 kg food and with a normal diet as control. Hematological parameters such as red blood cells (RBC) and white blood cells (WBC), hematocrit (Hct), hemoglobin (Hb), RBC index like mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) as well as immunological parameters such as peroxidase, lysozyme and complement activities, total protein, albumin and globulin levels were measured after 21 days of watercress extract treatment. The results indicated that oral administration of 1 % watercress extract in fish may enhance some hematological and immunological parameters including Hb and MCHC, lysozyme and complement activities, total protein and globulin levels, compared to the controls after 21 days of experimental period. In conclusion, on the basis of these results, oral administration of watercress extract may be useful to improve fish's immune system.

Transcript levels of five enzymes involved in cortisol synthesis and regulation during the stress response in common carp: relationship with cortisol.

In this study the expression of five genes involved in cortisol synthesis and regulation in the head kidneys of common carp (Cyprinus carpio L.) has been investigated in response to 3h net confinement stress, followed by 22h recovery. Cortisol, glucose, lactate and free fatty acid levels were measured in blood plasma. StAR, P450c17a2, 3betaHSD, P450c21 and 11betaHSD2 transcript sequences were identified based on Cyprinidae homologs and quantified by real-time PCR. Results showed that the plasma cortisol level reached a peak at one hour post-stress (85-fold higher than in control) and quickly returned to normal after 4h recovery. 11betaHSD2 transcripts were for the first time identified in interrenals. Changes in cortisol levels during and after confinement were correlated in a time-delayed relationship with increase and decrease in mRNA levels of 11betaHSD2, respectively. These results suggest that cortisol may be involved in the control or activation of 11betaHSD2. StAR and P450c21 mRNA levels did not change during net confinement stress and recovery, but P450c17a2 levels were significantly increased 4 and 22h after recovery. Since plasma cortisol levels increased by 68-fold within 5min net confinement stress, it seems that transcriptional activation of this enzyme is not directly involved in acute cortisol production.