The cytokine interleukin-1 (IL-1) has a variety of effects in brain, including induction of fever, alteration of slow wave sleep, and alteration of neuroendocrine activity. To examine the potential sites of action of IL-1 in brain, we used iodine-125-labeled recombinant human interleukin-1 [( 125I]IL-1) to identify and characterize IL-1 receptors in crude membrane preparations of mouse (C57BL/6) hippocampus and to study the distribution of IL-1-binding sites in brain using autoradiography. In preliminary homogenate binding and autoradiographic studies, [125I]IL-1 alpha showed significantly higher specific binding than [125I]IL-1 beta. Thus, [125I]IL-1 alpha was used in all subsequent assays. The binding of [125I]IL-1 alpha was linear over a broad range of membrane protein concentrations, saturable, reversible, and of high affinity, with an equilibrium dissociation constant value of 114 +/- 35 pM and a maximum number of binding sites of 2.5 +/- 0.4 fmol/mg protein. In competition studies, recombinant human IL-1 alpha, recombinant human IL-1 beta, and a weak IL-1 beta analog. IL-1 beta +, inhibited [125I]IL-1 alpha binding to mouse hippocampus in parallel with their relative bioactivities in the T-cell comitogenesis assay, with inhibitory binding affinity constants of 55 +/- 18, 76 +/- 20, and 2940 +/- 742 pM, respectively; rat/human CRF and human tumor necrosis factor showed no effect on [125I]IL-1 alpha binding. Autoradiographic localization studies revealed very low densities of [125I]IL-1 alpha-binding sites throughout the brain, with highest densities present in the molecular and granular layers of the dentate gyrus of the hippocampus and in the choroid plexus. Quinolinic acid lesion studies demonstrated that the [125I]IL-1 alpha-binding sites in the hippocampus were localized to intrinsic neurons. After hypophysectomy, homogenate binding and autoradiographic studies showed that there was no apparent change in the relative density of IL-1 receptors in the hippocampus. The identification of IL-1 receptors in brain with characteristics similar to IL-1 receptors in immune and neuroendocrine tissues provides further support for a physiological role for IL-1 to regulate central nervous system activity.

Background. Autophagy is an intracellular bulk degradation process induced by cell starvation. Autophagy was recently reported to be induced by various stresses such as hypoxia, ischemia/reperfusion, toxins, and denatured proteins, and to affect cell survival and death. Light chain 3-II (LC3-II) is specifically located on double membrane-bound autophagosomes that envelop disused proteins or organelles. Method. Transgenic mice in which green fluorescent protein (GFP) was fused to LC3(LC3-GFP) were administered cisplatin (20 mg/kg). After euthanasia at times between 0-72 hours, kidneys were excised for immunohistochemical analyses.Microscopic examinations of the generated NRK-52E cell lines stably transfected with LC3-GFP, and western blot analyses of NRK-52E cells were undertaken after cisplatin treatment with or without autophagy inhibitors and beclin 1 siRNA.Results. Autophagosomes increased in the proximal tubular cells of transgenic mice from 12 hours after cisplatin injection (20 mg/kg). The time course for this was faster than those for tubular necrosis and apoptosis. Autophagosomes also increased in NRK-52E cells after cisplatin treatment, with the time course for this faster than that for apoptosis. When autophagy was suppressed by autophagy inhibitors or beclin 1 siRNA, the level of apoptosis was also suppressed. Conclusion.Autophagy occurs in proximal tubular cells after cisplatin treatment and is involved in cell death in renal tubular injury. Our data suggested that autophagy is a kind of cell damage index and cells with activated autophagy will be scavenged by apoptosis. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 4 and in other cases induces apoptosis or type-2 programmed cell death [6][7][8]. Chronic myocardial ischemia/reperfusion injury induces autophagy increment and apoptosis decrement [9]. Inhibition of autophagy prevents neuron death after hypoxic-ischemic injury in neonatal mice [10]. However, in acute kidney injury (AKI), the association between autophagy and cell death is not obvious.Cisplatin is a major chemotherapeutic drug against solid tumors. One of the most important side effects of cisplatin is nephrotoxicity. Cisplatin is freely filtered at the Subjects and Methods MaterialsAnti-light chain 3 antibody was purchased from MBL (Nagoya, Japan).Anti-aquaporin-1 antibody was purchased from Abcam (Cambridge, MA).Anti-cleaved caspase 3 and rapamycin were purchased from Cell Signaling (Danvers, 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 Cell CultureThe NRK-52E cell line was obtained from ATCC (Manassas, VA), and cultured in DMEM medium containing 10% fetal bovine serum, 100 u...

During pregnancy, changes in circulating levels of hormones, including estrogens, correlates with a significant decrease in the relapse incidence in women with Multiple Sclerosis (MS). In the present study, we demonstrate that both primary and cell line cultures of rat oligodendrocytes express the estrogen receptor (ER)-a and ERb estrogen receptors in the cytosol and nucleus, and that nuclear compartmentalization becomes more pronounced as the cells mature. Moreover, 17b-estradiol significantly decreases the cytotoxic effects of the peroxynitrite generator 3-(4-morpholinyl)-sydnonimine (SIN-1) in both immature and mature oligodendrocytes in a dose dependent manner. This protective mechanism requires pretreatment with 17b-estradiol and is blocked by ICI 182,780, a selective ERa/ERb antagonist. These results strongly suggest that 17b-estradiol protects oligodendrocytes against SIN-1 mediated cytotoxicity through the activation of the estrogen receptors and provides new insights into the roles of the estrogen signaling pathways in myelin forming cells that are lost in demyelinating disorders.

The objective of this investigation was to examine the stress and coping styles in Japanese nursing students. The principal measures of the stress and coping styles were the General Health Questionnaire (GHQ)-12 and Brief Coping Orientations to Problems Experienced scale. In a cross-sectional analysis, 1324 students completed the anonymous self-administered questionnaires including the scales earlier. Feeling stress, living with family, not eating breakfast every day, having no regular exercise and poor sleep were associated with GHQ responder (psychological distressed group). The most commonly reported source of stress was taking examinations, followed by relationships with friends, engaging in clinical practice and presenting reports. The three most common coping styles adopted by the nursing students were acceptance, self-distraction and using instrumental support. By logistic regression analysis of coping styles with GHQ responder, self-blame, active coping, acceptance and behavioural disengagement were highly associated with GHQ responder. The nursing school educators as well as students should be aware of stress management strategies (e.g. using active coping and avoiding self-blame) that may help prevent depression.