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Purpose. The purpose of the study was to identify the relationship between plasma β-endorphin (objective variable) and the dimensions of the Chinese Cancer Pain Assessment Tool (CCPAT) (subjective variables). The dimensions included functional, pharmacological, psychosocial, pain beliefs and meanings, emotional and pain intensity. Methods. In this correlational study, 48 Chinese patients who had experienced cancer pain in the 24 hours before the investigation were recruited from an oncology unit by convenience. CCPAT scores and venous blood for β-endorphin were taken from each subject concurrently on two occasions, 8 a.m. and 8 p.m. Plasma β-endorphin was measured by radioimmunoassay. Results. Thirty-eight men and nine women were recruited. With the exception of 14 subjects who did not have the blood-taking procedure in the evening, all subjects completed the CCPAT and their blood was taken. Nineteen subjects suffered from gastrointestinal cancers, 17 from nasopharyngeal carcinoma, 9 from lung ca...

Doxorubicin (DOX) is commonly used for the treatment of hematological and solid tumors. However, there are serious toxic effects on the cardiovascular system, which limits the application of the drug. Recently, melatonin has been reported to have immunomodulatory effect in addition to lowering cholesterol levels as well as inhibiting malignant tumors. In this study, the effect of melatonin against the toxicity of doxorubicin was investigated in rats. Hemodynamic function, pathological and biochemical changes were determined in different treated hearts. Our findings showed that a significant protection by melatonin (6 mg kg -1 for 15 days, cumulative dose of 90 mg kg -1 ) against the DOX-induced toxicity was observed. Cardiac function was improved and lipid peroxidation decreased after melatonin treatment. It is concluded that melatonin provides protection against doxorubicin toxicity via an attenuation of lipid peroxidation.

Pang CS, Tang PL, Song Y, Pang SF, Ng KW, Guardiola-Lemaitre B, Delagrange P, Brown G.M. Differential inhibitory effects of melatonin analogs and three naphthalenic ligands on 2-[l25I]iodomelatonin binding to chicken tissues. J. Peneal Res. 1997; 23:148–155. © Munksgaard, Copenhagen Abstract We have compared the 50% inhibition values of 2-[l25I]iodomelatonin ([125I]Mel) competition curves by melatonin and 3 naphthalenic ligands, N-[2-(7-methoxy-l-naphthyl) ethyl] cyclobutane carboxamide (S20642), N-propyl N-[2-(7-methoxy-l-naphtyl) ethyl] urea (S20753), and N-[2-(7-methoxy-l-naphthyl) ethyl] crotonamide (S20750), using membrane preparations of four tissues (lung, spleen, brain, and kidney) of the chicken simultaneously. In retired breeders, we have demonstrated that the affinities of S20642 were similar in the lung and spleen. However they were 2-fold lower in the brain and 80-fold lower in the kidney. Similar differential binding affinities to the melatonin receptors were observed in the four tissues of young male chicks. This suggests that age and sex have little influence on the differential inhibitory properties of melatonin and S20642 in the tissues studied. The addition of guanosine 5'-0-thiotriphosphate (GTPyS), which encouraged the uncoupling of melatonin receptor to the G protein complex, lowered the binding affinity of melatonin and S20642 in the tissues studied but their differentia] affinities in the four tissues were however maintained. The affinities of 5-methoxy-N-cyclopropanoyltryptamine (CPMT) in the kidney were also 5–10-fold lower than those in the lung, spleen, and brain of young male chicks. The distinctive differential affinities of melatonin, S20642, and CPMT for [l25I]Mel binding sites in the chicken lung, spleen, brain, and kidney indicated that the binding sites in these tissues are heterogeneous. Our study implicated that the naphthalenic ligand S20642 may be a useful melatonin analog to distinguish melatonin receptor subtypes in tissues and a possible drug candidate worthwhile for further investigations.

The effect of superoxide dismutase (SOD) on membrane integrity and fluidity of the cultured neonatal rat cardiac myocytes in vitro was investigated under the condition of hypoxia and hypoxia/reoxygenation. Lactate dehydrogenase (LDH) concentration was used as the biochemical indicator for the loss of cell membrane integrity. Fluorescence polarization (FP), average microviscosity (eta) and anisotropy (Ast), which are inversely proportional to the fluidity of cell membrane, were assayed. Cells were respectively exposed to hypoxia or hypoxia/reoxygenation for different periods of time in the absence or presence of SOD at various concentrations. Hypoxia alone or hypoxia/reoxygenation brought injury to the cultured myocytes. This was demonstrated by changes in LDH and membrane fluidity. In the former LDH concentration gradually increased in a time-dependent manner and the values of FP, eta and Ast were significantly increased. The changes in membrane integrity and fluidity induced by hypoxia or hypoxia/reoxygenation could be prevented by adding SOD to the culture medium. The results provide a direct evidence that SOD (740 u.ml-1, the effective dose) was effective in protecting cultured myocytes against the injury as well as an indirect evidence of free radical generation. Based on the results obtained from this study and the establishment of concept of optimally effective dose by Bernier and Omar et al, it was suggested that some previous reports, in which no evidence was found both in protective effect of SOD and in free radical generation by using only one dose in hypoxia/reoxygenation model, should be interpreted with caution.

Experimental data of transferrin and transferrin-bound iron uptake byrabbit reticulocytes in the presence or absence of extracellular lead isanalyzed by means of a fractal model. A highly significant correlation offractal dimension (Df) of intracellular transferrin or transferrin-boundiron uptake with varying extracellular concentrations of lead (0 ~ 25umol/L) was observed (Transferrin: r = 0.897, p = 0.015; transferrin-boundiron: r = 0.947, p = 0.004). The Df of membrane-bound transferrin (r =-0.618, p = 0.191) or transferrin-bound iron (r = 0.144, p = 0.786) did notappear to be markedly altered by lead. Further analysis shows thatinhibitory degree of lead on intracellular iron uptake is higher than thaton intracellular transferrin uptake. These results suggest that theinhibitory effect of lead on the iron uptake may occur in intracellularprocess rather than in membrane binding step, probably inhibitingtranslocation of iron across the endosomal membrane.

The ability of melatonin to protect iron-induced lipid peroxidation was studied in various rat cell membranes. The concentration of cellular membrane malondialdehyde (MDA) was used as an index of induced oxidative membrane damage. Cell membranes from the brain, heart, kidney and liver of the male Sprague-Dawley rat were incubated with ferric ammonium citrate (20 microg/ml iron) alone for 3 h and concomitant with varying concentrations of melatonin ranging from 125 to 2,000 microM. The basal MDA levels of all the cell membranes were 25.0+/-1.4 (brain), 21.2+/-0.2 (heart), 10.0+/-0.9 (kidney) and 20.7+/-0.4 (liver) microM/g membrane protein, and the highest lipid peroxidation after exposure to iron occurred in the kidney (314.4%), followed by the heart (151.3%), the liver (130.4%) and the brain (121.7%). This peroxidative effect was completely (ED50 846.7 microM for the heart) and partially suppressed by melatonin (ED50 462.1 microM for the brain, 178.3 microM for the kidney and 886.6 microM for the liver). This inhibition effect on MDA production by these cell membranes was also found - except for the liver - if melatonin was used alone. These results show that the direct effect of lipid peroxidation on cellular membrane following iron exposure is markedly reduced by melatonin.

On the basis of the discussion in this paper, the process of transferrin and iron (transferrin-bound iron and non transferrin-bound iron) uptake and transferrin release by reticulocytes are summarized diagrammatically. Although we were able to outline the pathways shown in the figure, there is still a long way to go before we achieve total understanding of the mechanisms of iron uptake. In addition, many important questions need to be answered. For example: what is the nature and properties of the iron carrier on the membrane? What is the relationship between the iron carrier and transferrin receptor? Is the iron carrier system in membranes of cells from different tissues similar or different? And how does iron cross the membrane of the endosomes after it is released from transferrin? All of these questions merit further investigation.

We have compared the pharmacological characteristics of 2-[125I]iodo-melatonin binding to crude membrane preparations of the lung, spleen, brain and kidney of chicken. Saturation studies in

Classically, sensory receptors are specialized cells which detect specific environmental disturbances and send out neural signals for the integration, control and/or regulation of effector organs. Recently, a special class of sensory receptors called sensori-hormonal cells which employ hormones as their means of flux of biological information has been proposed. These sensori-hormonal cells are capable of detecting and transducing environmental signals directly into the secretion of hormones within the same cells. Theoretically, all sensory receptors may have examples capable of direct sensori-hormonal transduction. However, only one group of sensori-hormonal cells, the photoendocrine cells, have so far been studied. The photoendocrine cells including the photoreceptors of fish retinas and pinealocytes of bird pineals are capable of detecting light and/or darkness and transducing the electromagnetic radiation energy into a hormonal output. Generally, light suppresses and darkness stimulates the biosynthesis and secretion of melatonin by these photoendocrine cells. Contrary to many hormonal systems which employ principally the feedback mechanism for regulation, the sensori-hormonal cells are predominantly controlled by the feedforward mechanism. However, other factors may serve as additional means of regulation by influencing the system and affecting the transduction processes and/or the synthesis and secretion of the hormone. The ability of sensori-hormonal transduction is suggested to be important for the survival of the organism itself and/or its species and sensori-hormonal cells or their equivalent should appear early in the course of animal evolution. It is further suggested that the sequence of appearance of melatonin functions in the course of evolution would be: hormone--humoral factor--neuromodulator--neurotransmitter.

This paper presents findings on utilizing bright light to entrain the sleep–wake cycle in order to improve the perceived quality of sleep. The feasibility of including bright light as one of the nonpharmacologic nursing interventions for sleep promotion is discussed. A decade ago, nurses relied on sleeping pills to solve patients' sleep problems (Halfens et al. 1991). Reported obstacles to the development of sleep management tools included insufficient knowledge of sleep (Edell-Gustafasson et al. 1994) and unfamiliarity with effective nursing methods for tackling sleep problems (Halfens et al. 1991). Ten years later, nurses took the initiative in sleep promotion activities. In their book, Morgan and Closs (1999) introduced several sleep management methods in nursing practice. Floyd et al. (2000) summarized that 12 nonpharmacologic interventions used by nurses in the past to promote sleep. These 12 interventions are categorized according to their mechanism of action as relaxation, noise management, circadian repatterning and sleep hygiene. Light is thought to regulate the circadian system (Dawson & Van den Heuvel 1998) as a major synchronizer of endogenous circadian cycles (Dijk et al. 1995).

This study aims to determine the effects of light on the levels of 5-MIAA to provide further information on this indoleamine, using a sensitive and specific radioimmunoassay (RIA) for immunoreactive 5-methoxyindole-3-acetic acid (5-MIAA) developed in our laboratories using a specific antibody and tritiated label. Significant differences were found in the immunoreactive 5-MIAA levels between mid-light and mid-dark pineal glands in rats adapted to 12/12 hrs light/dark and in constant darkness. Under constant light, the circadian rhythm was abolished. The rat serum displayed no diurnal variations in 5-MIAA levels under aux photic conditions. The persistence of rhythms found in constant darkness but abolished in constant light suggests that the pineal 5-MIAA is endogenous and uses light as an environmental cue. In addition to melatonin, 5-MIAA could possibly be another pineal photoperiodic signal in animals.

We hypothesized that doxorubicin (DOX) induces cardiotoxicity of myocardium via oxygen radicals. The present study is aimed at examining the membrane alterations by oxygen radicals generated by DOX in adult rats and cultured neonatal myocytes. Our results showed that DOX 1) decreased beta-adrenoceptor (BAR) density in the cell membrane, 2) increased the membrane permeability of cultured neonatal rat myocytes and 3) altered the ultrastructure of myofibrils and subplasmalemmal actin networks. These effects were reproducible by exogenous hydrogen peroxide. The antioxidant melatonin (MLT) inhibited enzyme leakage and peroxidation in a concentration-dependent manner. It is concluded that DOX induces cardiotoxicity through lipid peroxidation and melatonin is an effective antioxidant against the reactive oxygen intermediates generated by DOX.

This study investigated the effects of strenuous exercise on transferrin (Tf)-receptor (TfR) expression and Tf-bound iron (Tf-Fe) uptake in erythroblasts of rat bone marrow. Female Sprague-Dawley rats were randomly assigned to either an exercise or sedentary group. Animals in the exercise group swam 2 h/day for 3 mo in a glass swimming basin. Both groups received the same amount of handling. At the end of 3 mo, the bone marrow erythroblasts were freshly isolated for Tf-binding assay and determination of Tf-Fe uptake in vitro. Tissue nonheme iron and hematological iron indexes were measured. The number of Tf-binding sites found in erythroblasts was ∼674,500 ± 132,766 and 1,270,011 ± 235,321 molecules/cell in control and exercised rats, respectively ( P < 0.05). Total Fe and Tf uptake by the cells was also significantly increased in the exercised rats after 30 min of incubation. Rates of cellular Fe accumulation were 5.68 and 2.58 fmol ⋅ 106cells−1⋅ min−1in the exercised and control rats, respectively ( P < 0.05). Tf recycling time and TfR affinity were not different in exercised and control rats. Increased cellular Fe was mainly located in the stromal fraction, suggesting that most of accumulated Fe was transported to the mitochondria for heme synthesis. The findings demonstrated that the increased cellular Fe uptake in exercised rats was a consequence of the increased TfR expression rather than the changes in TfR affinity and Tf recycling time. The increase in TfR expression and cellular Fe accumulation, as well as the decreased serum Fe concentration and nonheme Fe in the liver and the spleen induced by exercise, probably represented the early signs of Fe deficiency.

In this study, the mechanism of transferrin-free iron uptake by brain neuronal cells was investigated using the cultured cerebellar granule cells. Effects of incubation time, iron concentration, temperature and other divalent metals on the cellular uptake were determined. After five days of plating, the cells were incubated with different concentrations of transferrin-free iron in isotonic sucrose solution at different temperatures for a certain time. The cellular transferrin-free iron uptake was analysed by measuring the cellular radioactivity with a gamma-counter. The result showed that the cultured cerebellar granule cells had the capacity to acquire transferrin-free iron at pH 6.5, at which it was demonstrated that transferrin binds iron very poorly and only very little transferrin can be internalized by reticulocytes and HeLa cells. The iron uptake by cells increased with incubation time in a linear manner at a rate of 0.1076 pmol/microg protein/min within the first 10 min. The uptake was time- and temperature-dependent, iron concentration saturable, and inhibited by several divalent metal ions, such as Co2+, Zn2+, Mn2+ and Ni2+. These characteristics of transferrin-free iron uptake by the cultured cerebellar granule cells observed in this study, similar to those obtained from cells outside of the brain, implied that a carrier-mediated iron transport system might be present on the membrane of this type of brain neuronal cells. In addition, no significant difference in malondialdehyde measurement was found when the cells were incubated without or with the lower concentrations of iron (4 microM) for 20 min at 37 degrees C, demonstrating that this system was valid for studying membrane iron transport in this type of brain neuronal cell.

Excessive brain iron has been found in several neurodegenerative diseases. However, little information is available about mechanism of iron uptake by different types of brain cells including neurons. In this study, transferrin-bound iron (Tf-Fe) accumulation in the cultured cerebellar granule cell was investigated in vitro. After 5 days of culture, the cells were incubated with 1 microM of double-labelled transferrin (1251-Tf-59Fe) at 37 degrees C for 60 min. The cellular Tf-Fe and transferrin (Tf) uptake was analysed. The result showed (1) Tf uptake by the cells increased rapidly at the first 5 min, reaching its maximum after about 20 min of incubation; (2) Tf-Fe uptake kept increasing in a linear manner during the whole period of incubation; (3) the addition of either NH4Cl or CH3NH2, the blockers of Tf-Fe uptake via inhibiting iron release from Tf within endosomes, decreased the cellular Tf-Fe uptake but had no significant effect on Tf uptake; (4) trypsin and unlabelled Tf-Fe inhibited the uptake rate of Tf-Fe as well as Tf. The results suggested that Tf-Fe transport across the membrane of this type of neuron, much like other mammalian cells, was mediated by Tf-TfR endocytosis. Dysfunction of Tf or TfR would possibly lead to iron irregulation in the brain and consequently cause damage to neuronal functions.

Qian, Z. M., D. S. Xiao, W. Q. Huang, P. L. Tang and B. Xu. Central ANG II receptor involved in carotid sinus reflex resetting in chronically stressed rats. Physiol Behav 62(2) 241–247, 1997.—The performance of carotid sinus baroreceptor reflex (CSR) was characterized in chronically stressed rats by changing intracarotid sinus pressure (ISP) and constructing ISP-MAP (mean arterial pressure) relationship curve. The role of central angiotensin II (ANG II) receptors in the changes of CSR performance induced by chronic stress was determined. Rats were subjected to foot-shock stress for two weeks. The carotid sinus was isolated from the systemic circulation and the ISP changed in a stepwise manner. The results showed that in chronically stressed rats, ISP-MAP relationship curve shifted upward, the set point was significantly higher than that obtained from the unstressed group, and the reflex gain and the MAP range were significantly smaller than those in unstressed rats. After intracerebroventricular injection of saralasin (20 ng), MAP range was augmented and the set point decreased significantly. Injection of vehicle did not lead any significant differences between the parameters of the reflex measured before or after injection in either the stressed or the unstressed rats. Furthermore, administration of ANG II (10 μ g) induced a significant increase in the set point and decrease in the reflex gain in the unstressed rats. The responses of CSR to ANG II were completely blocked by pretreatment of saralasin. These findings suggest that chronic stress could induce the decreased CSR function in the normotensives and central ANG II receptors involved in the resetting of CSR in the chronically stressed rats.

Polysaccharide peptide (PSP) is a protein-bound polysaccharide extracted from an edible mushroom, Coriolus versicolor. Effects of PSP (2g/kg/day) on cyclophosphamide (CPA, 40 mg/kg/2 days)-induced immunosuppression were investigated by determining lymphocyte proliferation, Natural Killer (NK) cell function, IgG and IL-2 concentration, WBC count and the weight of organs after rats were treated with or without CPA in the presence or absence of PSP. The results demonstrated that PSP possessed immunopotentiating effect, being effective in restoring CPA-induced immunosuppression such as depressed lymphocyte proliferation, Natural Killer cell function, production of white blood cell and the growth of spleen and thymus in rats as well as in increasing both IgG and IL-2 production on which CPA did not have significant effects under the conditions of our experiments. PSP can partly restore CPA-induced immunosuppression. Based on our findings and the data accumulated so far, it was suggested that PSP should be considered as an useful adjuvant especially combined with CPA or other chemotherapy in clinical treatment of cancer patients. The mechanism by which PSP restores the immunosuppression induced by CPA is unclear.

The antioxidant and possible pro-oxidant effects of melatonin and related indoleamines (tryptophan, serotonin, N-acetylserotonin, and 5-methoxytryptamine) were studied in vitro. In two model membrane systems, i.e., phospholipid liposomes and rat erythrocytes, lipid peroxidation induced by Fe 2+ and H 2 O 2 , respectively, were reduced by the tested indoleamines except for tryptophan. The 5-hydroxy-indoleamines, serotonin, and N-acetylserotonin exhibited pro-oxidant actions in the bleomycin assay by reducing Fe 3+ to Fe 2+ , which leads to DNA damage. The 5-methoxy-indoleamines, melatonin and 5-methoxytryptamine, were devoid of any pro-oxidant actions in this assay. Serotonin, but not N-acetylserotonin, scavenged the superoxide anion. None of the indoleamines tested had any reactivity towards H 2 O 2 . All the indoleamines, including tryptophan, were, however, shown to react with hypochlorous acid. These findings support an antiperoxidative and antioxidant action of melatonin which is devoid of pro-oxidant effect on non-lipid substrates.

To investigate whether melatonin has a direct action on the cardiovascular system, putative melatonin receptors were studied in quail heart membrane preparations using the specific melatonin agonist 2-[125I]iodomelatonin ([125I]Mel) as the radioligand. The [125I]Mel binding demonstrated in the mature quail heart was saturable, highly specific and reversible, of picomolar affinity and femtomolar density (Kd = 35.2 ±5.2 pM; Bmax = 1.32 ± 0.25 fmol/mg protein; n = 8). The linear Scatchard plots and the close to unity Hill coefficient indicated a single class of binding sites. The pharmacological profile was in the affinity order of 2-iodomelatonin = 2-phenylmelatonin > melatonin > 6-chloromelatonin ⪢ 6-hydroxymelatonin > 6-sulphatoxymelatonin ⪢ N-acetylserotonin ⋙ 5-hydroxytryptamine. Guanosine 5′- riphosphate and guanosine 5'-O-(3-thiotriphosphate) (GTPγS) dose dependently inhibited the binding. Ten μM GTPγS lowered the binding affinity by 50% in saturation studies. The order of potency of inhibition by cations was: Ca2+ > Mg2+ > Li+ > Na+ > K+ > choline chloride. Contrary to most other melatonin binding sites, millimolar concentrations of Ca2+ and Mg2+ did not promote binding in the quail heart membranes. In vitro autoradiography indicated homogenous labeling in the heart. Our results demonstrated [125I]Mel binding sites in the quail heart. That guanine nucleotides and Na+ inhibited the binding indicated that these putative melatonin receptors are coupled to guanine nucleotide-binding proteins (G-proteins).

[ 125 I]Iodomelatonin binding was first demonstrated in the chicken lung membrane preparations. The binding was saturable, reversible, rapid, and of high affinity. The sites of binding distributed widely in different subcellular fractions except the cytosol fraction. Scatchard plots are linear indicating a dissociation constant ( K d ) of 8.11 ± 0.73 pmol/l and a maximum number of binding sites of 1.29 ± 0.16 fmol/mg protein ( n = 9). The K d estimated from the kinetic study was 11.2 pmol/l. Guanosine 5′- O -(3-thiotriphosphate) significantly increased the K d values while the density of binding sites was not affected indicating that the binding sites may be linked to a guanine nucleotide binding protein.

The relationship between lipid peroxidation and uptake of transferrin- free iron, Fe(II), by reticulocytes in an experimental system for studying membrane transport of Fe(II) was investigated by using free radical scavengers: BHA (butylated hydroxyanisole), BHT (butylated hydroxytoluene), superoxide dismutase, alpha-tocopherol, propyl gallate and DPPD (N,N-diphenyl-1,4-phenylenediamine), and producers: t-butyl hydroperoxide, cumene hydroperoxide, H2O2 and aluminium carbonate. Measurements were made of MDA (malondialdehyde) and the rate of Fe(II) uptake from a sucrose solution buffered at pH 6.5 by Pipes. Most scavengers and producers used could increase or decrease only slightly the rate of Fe(II) uptake and some of them had no effect on Fe(II) uptake and MDA could not be detected at iron concentration of lower than 10 microM and incubation time of 20 min. At iron concentration of higher than 100 microM and incubation time of 4 h, there was the production of MDA which increased with the increment of iron concentration of incubation medium and BHT could inhibit the production of MDA. In addition, no difference was found in the rates of Fe(II) uptake in three experimental groups whose incubation medium was buffered by Pipes, Mops and Mes respectively. The results suggested that iron could induce free radical reaction under experimental conditions, especially at high concentration of iron and longer incubation time; however, at low concentration of iron (10 microM) and the usual incubation time (20 min) free radical reaction was very slight and the extent of the reaction was not enough to damage the integrity and function of the membrane of reticulocytes, and that Fe(II) uptake by reticulocytes was not the result of free radical reaction and lipid peroxidation. It was therefore concluded that iron could not initiate its own membrane transport in rabbit reticulocytes by free radical reaction and lipid peroxidation and that the experimental system we used for studying membrane transport of Fe(II) is valid.