Effects of dietary Spirulina on vascular reactivity.

There are several reports suggesting that Spirulina (Arthrospira) may have a beneficial effect in the prevention of cardiovascular diseases. Here we review the results of studies on the effects of dietary Spirulina on the vasomotor reactivity of aortic rings excised from either lean or obese Wistar rats. We also review preliminary results on the effects of Spirulina intake on plasma lipids and blood pressure in humans. The results of the former studies strongly suggest that Spirulina induces a tone-related increase in the synthesis/release of nitric oxide by the endothelium as well as an increase in the synthesis/release of a vasodilating cyclooxygenase-dependent metabolite of arachidonic acid and/or a decrease in the synthesis/release of a vasoconstricting eicosanoid by the endothelium. In humans, Spirulina maxima intake decreases blood pressure and plasma lipid concentrations, especially triacylglycerols and low-density lipoprotein-cholesterol, and indirectly modifies the total cholesterol and high-density lipoprotein-cholesterol values.

Effects of single sessions of low-intensity continuous and moderate-intensity intermittent exercise on blood lipids in the same endurance runners.

The factors responsible for the acute effects of exercise on blood lipids are not well known, and there have been few studies comparing different kinds of exercise in the same population. The concentration of blood lipids was evaluated in this study at the end and at post-24h of two 14km/90min single exercise sessions: continuous exercise (CE) at 44.5+/-5.6% VO(2max) and intermittent exercise (IE) at 39-72% VO(2max), in subjects with high levels of aerobic training. Fourteen male athletes (endurance runners) took part in this study and each completed a 24h dietary record. The O(2) uptake and CO(2) production were recorded, and blood lactate and blood lipids were measured. The results showed that triacylglycerols were not modified by any kind of exercise. Total cholesterol was increased at the end of both exercises: 7.04% for CE (p<0.001) and 4.23% for IE (p=0.001). High-density lipoprotein cholesterol was increased at the end of IE: 11.38% (p=0.03) and low-density lipoprotein cholesterol was increased only at the end of CE: 7.45% (p=0.006). The increase of lipids for CE was negatively correlated with aerobic fitness indicators (heart rate and %HRmax at lactate threshold), and was positively associated with energy expenditure. For IE, %HRmax and lactate were negatively correlated, and the respiratory exchange ratio was positively correlated, with the lipid increase. We conclude that in trained male athletes, a 14km run in 90min induced different changes of lipid profile if the exercise was done continuously or intermittently, and that in CE the extent of these increases was influenced by aerobic fitness.

Relationship between carnitine, fatty acids and insulin resistance.

Increased plasma free fatty acid (FFA) levels are a feature of insulin resistance and type 2 diabetes. The aim of the present study was to assess the effect of L-carnitine supplementation on plasma lipids and the expression of enzymes in peripheral mononucleated cells (PMNC) involved in the regulation of fatty acid and glucose oxidation. L-Carnitine supplementation of 2 g/day resulted in a significant decrease in plasma FFA and in a less pronounced diminution of the plasma triacylglycerols. In addition, a concomitant increase in the relative mRNA abundances of carnitine acyltransferases (5- to 10-fold) and of the carnitine carrier OCTN2 (12-fold) in PMNC of pregnant women was found. The results of the present study provide evidence that L-carnitine supplementation in pregnancy (2 g/day) avoids a striking increase in plasma FFA, which are thought to be the main cause of insulin resistance and consequently gestational diabetes mellitus.

Effects of testosterone and growth hormone on long-term retention and extinction of a passive avoidance response in young and aged rats.

Wistar rats were divided into two blocks: young and aged. Each block was then randomly divided into the following groups: (1) Control (saline solution), (2) Growth Hormone (GH), (3) Testosterone enanthate, and (4) GH plus testosterone enanthate. Animals were trained on a one-trial passive avoidance conditioning and tested for retention 24 h after training and thereafter twice a week until the extinction response occurred. Results showed that GH, testosterone, and GH plus testosterone improved long-term memory in young rats. Both GH and testosterone delayed the extinction process in young and aged rats. The GH-testosterone interaction delayed the extinction only in young rats.

Monitoring of carbamazepine concentrations in wastewater and groundwater to quantify sewer leakage.

Monitoring of carbamazepine concentrations in wastewater and groundwater enables us to identify and quantify sewer exfiltration. The antiepileptic drug carbamazepine is hardly removed in wastewater treatment plants and not or just slightly attenuated during bank infiltration and subsoil flow. Concentrations in wastewater are generally 1000 times higher than the limit of quantification. In contrast to . many other wastewater tracers carbamazepine is discharged to the environment only via domestic wastewater. The results from this study carried out in Linz, Austria indicate an average exfiltration rate of 1%, expressed as percentage of the dry weather flow that is lost to the groundwater on the city-wide scale. This rate is lower than sewage losses reported in most other studies which attempted to quantify exfiltration on the basis of groundwater pollution. However, it was also possible to identify one area with significantly higher sewage losses. This method seems to be very suitable for the verification of leakage models used to assess sewer exfiltration on a regional scale.

Quantification of sewer exfiltration using the anti-epileptic drug carbamazepine as marker species for wastewater.

The anti-epileptic drug carbamazepine was used as marker species in wastewater to identify and quantify sewer exfiltration. In several studies carbamazepine turned out to be hardly removed in wastewater treatment and not or just slightly attenuated during bank infiltration. Concentrations in wastewater are generally 1000 times higher than the limit of quantification. In contrast to many other marker species a "young" drug as carbamazepine is discharged to the environment only by wastewater. The results from this study carried out in Linz, Austria indicate an average exfiltration rate, expressed as percentage of the dry weather flow that is lost on the city-wide scale, of 1%. This rate is lower than sewage losses reported in most other studies which attempted to quantify exfiltration on the basis of groundwater pollution. However, it was also possible to identify one area with significant higher sewage losses.

Endothelium-dependent effects of the ethanolic extract of the mistletoe Psittacanthus calyculatus on the vasomotor responses of rat aortic rings.

The mistletoe Psittacanthus calyculatus (Loranthaceae) is used in Mexican traditional medicine for the treatment of hypertension. In the present study the effects of a crude ethanolic extract of this mistletoe, on the vasomotor reactivity of superfused rat aortic rings (with or without a functional endothelium) were analyzed. Either in the absence or in the presence of L-NAME or indomethacin, the extract (12.5-800 microg/ml) had no effect on the basal tone of both types of rings. In phenylephrine-precontracted rings, low concentrations of the extract (up to 300 microg/ml) induced a small additional tension development in both types of rings; however, the tension increase was slightly larger in rings having an intact endothelium. At higher concentrations (400-800 microg/ml), the extract relaxed, concentration-dependently, phenylephrine-precontracted rings with an intact endothelium. This relaxation was completely reverted by the addition of L-NAME. When the extract was applied in the continuous presence of L-NAME to phenylephrine-precontracted rings, instead of a relaxation a marked additional tension development occurred. Indomethacin did not modify the relaxation induced by the extract. The results indicate that the ethanolic extract of this mistletoe induces, predominantly, an endothelium-dependent relaxation which seems to be mediated by the synthesis/release of nitric oxide.

Effects of different ozone doses on memory, motor activity and lipid peroxidation levels, in rats.

Ozone is one of the main atmospheric pollutants. Its inhalation causes an increase in free radicals, when these free radicals are not compensated by antioxidants, it leads to an oxidative stress state. This oxidative stress state has been implicated in neurodegenerative processes. To determine the effects of oxidative stress caused by exposure to ozone on memory and motor activity, we used 120 male Wistar rats exposed to one of the following ozone doses, (0.0, 0.1, 0.4, 0.7, 1.1 and 1.5 ppm), for four hours. After ozone exposure, short and long term memory of a one trial passive avoidance test were measured, and motor activity was registered for five minutes, in 10 rats of each group. In 16 rats exposed to 0.0, 0.4, 0.7 or 1.1 ppm lipid peroxidation levels from frontal cortex, hippocampus, striatum and cerebellum, were measured. Results show that ozone, causes memory impairment from doses of 0.7 ppm, decrease in motor activity from doses of 1.1 ppm, and increase in lipid peroxidation levels from doses of 0.4 ppm. that increase with the dose.

Design, synthesis, and modeling of novel cyclic thrombin receptor-derived peptide analogues of the Ser42-Phe-Leu-Leu-Arg46 motif sequence with fixed conformations of pharmacophoric groups: importance of a Phe/Arg/NH2 cluster for receptor activation and implications in the design of nonpeptide thrombin receptor mimetics.

The novel cyclic analogues cyclo(Phe-Leu-Leu-Arg-epsilonLys-Dap) (1) and cyclo(D-Phe-Leu-Leu-Arg-epsilonLys-Dap) (2), which differ only in the absolute conformation of Phe, have been designed and synthesized based upon the minimal peptide sequence Phe-Leu-Leu-Arg which has been found to exhibit biological activity for the thrombin receptor. Compound 1, in which all amino acids have the L-configuration, exhibited higher activity in the rat aorta relaxation and rat longitudinal muscle bioassays compared to compound 2, in which the Phe residue is in the D-configuration. This is attributed to the spatial proximity of the Phe and Arg in compound 1 which does not exist in its diastereomeric compound 2, as is depicted from a combination of NMR studies and computational analysis. Structure-activity studies (SAR) showed that the Phe and Arg side chains along with a primary amino group form an active recognition motif that is augmented by the presence of a second primary amino group in the cyclic peptide. We suggest that a comparable cyclic conformation may be responsible for the interaction of linear TRAPs with the thrombin receptor. The validity of this proposition was tested by the synthesis of four active nonpeptide thrombin receptor mimetics. Substance (S)-N-(6-guanidohexanoyl)-N'-(2-amino-3-phenylpropionyl)piperazine (3), in which the pharmacophoric phenyl, guanidino, and amino groups were incorporated onto a piperazine template, was found to be the most active compared to the other synthesized compounds which lack the amino pharmacophoric group.

Effects of the ethanolic extract of Spirulina maxima on endothelium dependent vasomotor responses of rat aortic rings.

Dietary Spirulina decreases, endothelium-dependently, the responses to vasoconstrictor agonists and increases the endothelium-dependent, agonist-induced, vasodilator responses of rat aorta rings. The aim of this study was to analyze, in vitro, the effects of a raw ethanolic extract of Spirulina maxima on the vasomotor responses of rat aortic rings to phenylephrine and to carbachol. On rings with endothelium, the extract produced the following effects: (a) a concentration-dependent (60-1000 microg/ml) decrease of the contractile response to phenylephrine; (b) a rightward shift and a decrease in maximal developed tension, of the concentration--response curve to phenylephrine; (c) a concentration dependent relaxation of phenylephrine-precontracted rings. These effects were blocked by L-NAME, and not modified by indomethacin. The extract had no effect on the concentration-response curve to carbachol of rings with endothelium. On endothelium-denuded rings the extract caused a significant rightward shift of the concentration response curve to phenylephrine without any effect on maximal tension development. In the presence of the extract, indomethacin induced a marked decrease in the maximal phenylephrine-induced tension of endothelium-denuded rings. These results suggest that the extract increases the basal synthesis/release of NO by the endothelium and, also, the synthesis/release of a cyclooxygenase-dependent vasoconstricting prostanoid by vascular smooth muscle cells.

High-performance liquid chromatographic-tandem mass spectrometric determination of 3-hydroxypropylmercapturic acid in human urine.

A sensitive and specific high-performance liquid chromatographic-tandem mass spectrometric (HPLC-MS-MS) method was developed for the determination of 3-hydroxypropylmercapturic acid (3-HPMA) in human urine. Samples were extracted using ENV+ cartridges and then injected onto a C8 Superspher Select B column with acetonitrile and formic acid as eluent (5:95, v/v). N-Acetylcysteine was used as internal standard for HPLC-MS-MS. Linearity was given in the tested range of 50-5000 ng/ml urine. The limit of quantification was 50 ng/ml. Precision, as C.V., in the tested range of 50-5000 ng/ml was 1.47-6.04%. Accuracy ranged from 87 to 114%. 3-HPMA was stable in human urine at 37 degrees C for 24 h. The method was able to quantify 3-HPMA in urine of non-smokers and smokers.

Studies on the preventive effect of Spirulina maxima on fatty liver development induced by carbon tetrachloride, in the rat.

The aim of the present work was to assess if the feeding of either the oil extract of Spirulina maxima or of its defatted fraction would prevent fatty liver development, induced in rats by a single intraperitoneal dose of carbon tetrachloride (CCl4). Liver and serum lipids were evaluated 4 days after treatment with this agent. Concentration of liver lipids did not differ in rats fed on a purified diet either without or with one of the fractions of Spirulina, except for total cholesterol, which showed a slight increase in the group receiving the oil extract of Spirulina. However, after CCl4 treatment, liver total lipids and triacylglycerols were significantly lower in rats fed on a diet containing any fraction of Spirulina (defatted or the oil fraction) than in rats without Spirulina in their diet. Furthermore, the increased liver cholesterol values, induced by CCl4 treatment, were not observed in rats receiving Spirulina. In addition, rats receiving whole Spirulina in their diet and treated only with the vehicle showed an increase in the percentage of HDL values. The changes in VLDL and LDL induced by CCl4 treatment were not observed in the whole Spirulina group. Furthermore, after CCl4 treatment the values of the liver microsomal thiobarbituric acid-reactive substances were lower in the whole Spirulina group than in the control group. These results support the potential hepatoprotective role of Spirulina.

Spirulina maxima prevents induction of fatty liver by carbon tetrachloride in the rat.

The aim of the present work was to assess the capacity of Spirulina maxima to prevent fatty liver development induced in rats by an intraperitoneal single dose (1 ml/kg) of carbon tetrachloride. Liver and serum lipids were quantified two or four days after treatment with this agent. Liver lipid concentration did not differ in rats fed on a purified diet with or without Spirulina. However, after carbon tetrachloride treatment, liver triacylglycerols were significantly lower in rats fed on a diet with Spirulina 5% than in rats without Spirulina in their diet (P < 0.05). Furthermore, the increased liver cholesterol values, induced by carbon tetrachloride treatment, were not observed in rats that received Spirulina. These results support the potential hepatoprotective role of Spirulina.

Effects of dietary Spirulina maxima on endothelium dependent vasomotor responses of rat aortic rings.

The aim of this study was to evaluate the effects of Spirulina maxima on vasomotor responses of aorta rings from male Wistar rats fed on a purified diet. For this purpose, the animals (weighing 200-240 g) were allocated randomly in two groups. One receiving purified control diet (A) and the other receiving purified diet containing 5% Spirulina (B). Purified diets were according to American Institute of Nutrition guidelines and adjusted to Spirulina protein content. All animals were fed (20 g/day/rat) during two weeks, receiving water ad libitum and 12 h. light-dark cycles. Spirulina maxima effects were evaluated by concentration-response (CR) curves of aorta rings with or without endothelium to phenylephrine (PE), both in presence and absence of indomethacin (Indom) or indomethacin plus L-NAME (Indom. + L-NAME), and to carbachol (CCh). Aorta rings with endothelium from group B showed, relative to corresponding rings from group A: 1) a significant decrease in the maximal tension developed in response to PE. 2) this decrease was reverted by Indom. 3) Indom. + L-NAME induced an additional increase in the contractile responses to PE. 4) a significant shift to the left of the CR curve to CCh. No significant differences were observed in the tension developed in response to PE in rings without endothelium from either group. These results suggest that Spirulina maxima may decrease vascular tone by increasing the synthesis and release of both a vasodilating cyclooxygenase-dependent product of arachidonic acid and nitric oxide, as well as by decreasing the synthesis and release of a vasoconstricting eicosanoid from the endothelial cells.

Effects of acute and chronic estrogenic treatment on vasomotor responses of aortic rings from ovariectomized rats.

The effects of either chronic or acute estrogenic treatment on the "in vitro" vasomotor responses to phenylephrine (10(-9)-10(-5) M) and to carbachol (10(-9)-10(-5) M) of aortic rings excised from ovariectomized rats were analyzed. Chronic estrogenic treatment consisted in a single subcutaneous dose of 1 mumol estradiol 17-stearate. Effects of acute estrogenic treatment were evaluated by recording the responses of aortic rings excised from untreated ovariectomized rats both before and after the addition of 17 beta-estradiol to the superfusing solutions. In order to identify the endothelium-dependent responses each experiment was performed simultaneously on pairs of rings from the same aorta, one with and the other without functional endothelium. The contractile responses to phenylephrine of endothelium-intact vessels were attenuated by chronic estrogenic treatment; this attenuation was further increased by preincubation of the vessels with indomethacin and was reverted by N omega-nitro-L-arginine methyl ester. Either chronic or acute estrogenic treatment enhanced the carbachol-induced endothelium dependent relaxation of phenylephrine-precontracted rings. The results may be explained by assuming that estrogens increase the basal release of both nitric oxide and a cyclooxygenase-dependent vasoconstricting prostanoid as well as the receptor-mediated release of nitric oxide from the endothelium of the rat aorta.

The two components in the shortening of unloaded ventricular myocytes: their voltage dependence.

In isolated myocytes from mammalian ventricles a fast and a slow component in the contractile response to depolarizing voltage clamp steps were identified. The potential dependence of the slow component was identical to the activation curve of iCa. The fast component, however, remained at its maximal amplitude at potentials positive to +10 mV (up to +100 mV), in which potential range iCa declined and eventually disappeared. The results suggest that the slow component may be activated by Ca++ entering through sarcolemmal Ca channels, whereas the fast component depends on Ca release from intracellular sites and may depend on both Cai and voltage.

Recovery of the membrane potential of the sectioned striated muscle fibers of the guinea-pig cremaster.

We investigated changes in membrane potential after small transections (2-4 mm) of the striated muscle of the guinea-pig cremaster and the rate of penetration of procion yellow (1-5 mg/ml) into the injured fibers. Transverse severance of the striated muscle fibers was followed by a marked drop of the membrane potential near the site of injury, such a fall being undetectable about 2 mm from the lesion. The depolarization reached a peak value (40 mV at 0.5 mm from the lesion) 3-5 min after injury and was followed by a gradual repolarization which was complete in about 60 min. A second lesion evoked the same changes, but under conditions of a low calcium solution, there was no recovery in the potential. The rate of indicator penetration reached a peak value (20 micrometers/min) when depolarization was near maximal, declined as the membrane repolarized and became negligible in about 50 min. Conversely, the transected striated muscle fibers of the guinea-pig diaphragm remained depolarized for more than 1 hr after injury. These observations suggest that the healing-over process is a property of the striated muscle fibers of the guinea-pig cremaster and may tentatively be ascribed to the development of a calcium-dependent diffusion barrier (of unknown nature) in the area of the injury.