Buy Ecstasy Leuven

__________________________

📍 Verified store!

📍 Guarantees! Quality! Reviews!

__________________________

▼▼ ▼▼ ▼▼ ▼▼ ▼▼ ▼▼ ▼▼

▲▲ ▲▲ ▲▲ ▲▲ ▲▲ ▲▲ ▲▲

Buy Ecstasy Leuven

Tina Van Havere: 'The price of a gram of cocaine has been 50 euros for years now and an XTC pill will set you back 4 to 6 euros. Tina Van Havere: 'That prices are so low in Belgium has to do with the presence of the port of Antwerp. Many drugs enter the country via our ports and are then transported across the globe. While a gram of cocaine costs 50 euros in Belgium in Australia the price is apparently euros. XTC prices abroad can be ten times that in Belgium. Cannabis will set you back 5 euros for a joint. Scientists say that the extremely low prices in Belgium mean that loads of youngsters are experimenting with drugs. The price of an XTC pill is often cheaper than an alcoholic cocktail. Lawyer Tanja Smit, who is often involved in drugs cases, concedes that the low prices mean that there are few obstacles to accessing drugs with all the consequences this has. Headlines Categories vrtnieuws. Home News Charleroi Airport cancels all departures scheduled for Tuesday due to industrial action by security staff. Home News Indian summer beckons following Monday's downpours. Home News 'Little girl playing with pussy': Child Focus stepping up battle against child porn via explicit video titles. Sun 18 Nov Home News Amateur footballer seriously injured after a firework thrown onto the pitch by a supporter explodes in his hand. Home News September was a record month for traffic jams in Flanders. Home News Far-right Vlaams Belang joins a municipal coalition for the first time. Home News More than 30, demonstrators call for ceasefire in Lebanon and Gaza. Sun 20 Oct

XTC exhibition in Ghent: “Get people to think about alternative drug policy”

Buy Ecstasy Leuven

Official websites use. Share sensitive information only on official, secure websites. We aimed to assess in rats the acute MDMA toxicity to the brain and peripheral organs using a binge dose scheme that tries to simulate human adolescent abuse. After 24 h animal sacrifice took place and collection of brain areas cerebellum, hippocampus, frontal cortex and striatum and peripheral organs liver, heart and kidneys occurred. No brain oxidative stress-related changes were observed after MDMA. MDMA-treated rat organs revealed significant histological tissue alterations including vascular congestion, but no signs of apoptosis or necrosis were found, which was corroborated by the lack of changes in plasma biomarkers and tissue caspases. In peripheral organs, MDMA did not affect significantly protein carbonylation, glutathione, or ATP levels, but liver presented a higher vulnerability as MDMA promoted an increase in quinoprotein levels. Adolescent rats exposed to a moderate MDMA dose, presented hyperthermia and acute tissue damage to peripheral organs without signs of brain oxidative stress. In Europe, 2. Recreational use of MDMA is frequently done by binge administration that is a pattern of several administrations over a short time period \[ 4 — 6 \]. MDMA is a drug frequently used by adolescents and therefore we need to understand the consequences of MDMA exposure at this age through the administration of doses and schemes more close to the human situation. Interestingly, the vast majority of studies tend to use adult animals for studying MDMA toxicity. MDMA administration to adult laboratory animals was shown to promote long-term depletion of monoamine neurotransmitter content, damage to the nerve terminals, neuronal cell death and long-lasting cognitive impairments \[ 1 , 2 \]. Notably, studies conducted with adolescent animals have shown that MDMA exposure to neurotoxic doses leads to deficits in the serotonergic system \[ 2 \] and also late changes in memory and learning abilities \[ 7 , 8 \]. Investigation on adolescent laboratory animals is of extreme importance, given that studies with human adolescents have ethical barriers, making extremely difficult to enrol this population in studies that evaluate drug abuse toxicity. Nonetheless, studies with young adults reported deficits in the serotonergic system \[ 9 , 10 \], decreased grey matter concentration \[ 11 \] and cognitive deficits \[ 12 , 13 \] following MDMA abuse. These high doses, while important for studying neurotoxic related events, do not match the typical MDMA user profile, as they tend to be extremely high and correlate only to a high-intensity abuser. This method does not account for the differences in MDMA metabolism or administration route between humans and rats, but is certainly of great value for an approximate extrapolation. According to the latest European Union report MDMA pills range from 57 to mg \[ 15 \], and therefore that dose would mean an average intake of more than seven pills in a single session, a rather extreme scenario. We have conducted experiments that proved that this MDMA regimen did not induce serotonergic toxicity 7 days following MDMA administration to adolescent PND 40 Wistar rats, as no 5-HT depletion could be found in any of the four brain areas evaluated manuscript being prepared for submission. Therefore, and in accordance with the last European Union report on drugs, we estimate that the dose used in our protocol is equivalent to the intake of two to three pills by adolescents using the binge-dosing pattern. Accordingly, the current paradigm of exposure to adolescent animals tries to mimic the dose schedule used by human adolescents. For that reason studies using more moderate doses that closely match the typical MDMA user are needed. Additionally, MDMA abuse was associated with histopathological evidences of toxicity to the liver, heart and kidneys in humans \[ 24 , 25 \]. Studies are lacking regarding the evaluation of MDMA-induced damage to organs following lower doses. Moreover, at this point, studies in adolescent animal models regarding peripheral toxicity using moderate MDMA doses have not been done. To the best of our knowledge, this is the first study in an adolescent rat model to evaluate simultaneously in three peripheral organs oxidative stress parameters and histological damage. Folin—Ciocalteu reagent, cupper II sulphate, dimethyl sulfoxide, disodium phosphate, ethylenediaminetetraacetic acid EDTA , perchloric acid, potassium bicarbonate KHCO 3 , sodium hydroxide, sodium carbonate, magnesium chloride, potassium dihydrogen phosphate and magnesium sulphate were purchased from Merck Darmstadt, Germany. The fluorescent peptide substrates for the caspase activity assays were acquired from Peptanova Sandhausen, Germany. Animals were housed in a controlled environment \[temperature of Animals had ad libitum access to food and water and throughout the experimental period had permanent veterinary supervision. All procedures were performed as to give the proper animal care, to reduce suffering and stress. Then each animal was subjected to a brief inhalatory anaesthesia with isoflurane to perform a subcutaneous insertion of a temperature transponder BioMedic Data Systems Inc with minimum animal discomfort. This transponder ensures precise core body temperature measurements throughout the entire experimental period, as we reported before \[ 27 \]. Prior to MDMA administration, animals were maintained in groups allowing conspecific social interactions. In the administration day and for the next 24 h, animals were housed individually. We selected this sample size per group based on a pilot study that we performed. For a full explanation on these statistical calculations please see reference \[ 28 \]. Additionally, sample size took into consideration ethical reasons related to animal welfare in a pre-clinical study conducted with drugs of abuse, such as the present one. MDMA was prepared at the day of use in a concentration of 2. Control animals received saline solution at the same schedule and using equivalent injection volumes of treated animals. Food and water consumption, as well as animal weight were evaluated before the first injection and at the next day. Twenty-four hours following the first MDMA administration adolescent rats were sacrificed. Animals were anesthetized and euthanized with the volatile anaesthetic, isoflurane. Before decapitation, blood was collected from the inferior vena cava. Following sacrifice, brain areas cerebellum, hippocampus, frontal cortex and striatum and peripheral organs liver, heart and kidneys were collected. The collection of the different brain regions was performed in agreement to a rat brain atlas \[ 29 \], and the dissection techniques were conducted in accordance to a previous work \[ 30 \]. The brain and peripheral organs were also weighted. Samples of brain areas from the right hemisphere were collected in RIPA buffer supplemented with protease inhibitors \[0. The supernatants were separated for the protein carbonylation and quinoprotein analysis. The sample pellets were stored for protein determination. A section of liver, heart and kidneys was collected for caspases activity assay in caspase lysis buffer 0. Another section was collected in complete RIPA buffer supplemented with protease inhibitors, and homogenized using a sonicator 30 seconds, continuously , while tubes were kept on ice. Another section of each organ was collected and homogenized in an Ultra-Turrax samples diluted in 0. ATP levels were quantified by a bioluminescent assay using the luciferin-luciferase system, as described in detail in previous works \[ 26 , 32 \]. The results are presented in nmol of ATP per mg of protein. Protein carbonylation was determined as we previously reported \[ 33 \], with minor modifications. Samples 0. Incubation at room temperature with the secondary antibody anti-rabbit IgG-peroxidase, , 1 h was followed. Sections of liver, heart, and kidneys were prepared, and analysed, as previously described by our group \[ 31 \]. A fluorescent assay for tissues was used to determine each caspase activity in the liver, heart and kidneys of the animals, as previously described by our group \[ 31 \]. For other samples, proteins were quantified by the Lowry method \[ 34 \]. The Shapiro—Wilk normality test was conducted before group comparison. For data where two groups were compared, the t-test was used for a normal distribution or the Mann-Whitney Rank Sum test when data did not follow a normal distribution. Statistical analysis of the temperature, included in Fig. Statistical significance was accepted at p values less than 0. Temperature monitoring of adolescent rats after exposure to three doses of NaCl 0. MDMA-treated during 7 h. Temperature persisted significantly higher for almost 2 h after the third dose in treated animals. The temperature was also measured 24 h post-MDMA binge administration and no differences were observed in the temperature among controls and MDMA-treated rats data not shown. Recordings of body weight gain, food or water intake before and 24 h after exposure showed no differences among control and MDMA-treated rats data not shown. In fact, there was a slight decrease in the body weight of animals in both groups, possibly as a result of the animal stress due to the manipulation. The food consumption was similar in both control and MDMA-treated animals. However, there was a tendency for an increased water intake in animals that received MDMA, but it was not statistically significant data not shown. Each core organ liver, heart, kidneys, and brain had their weight registered and the weight ratio of each organ was taken to brain weight. No significant differences were observed between MDMA-treated rats and the control group for all collected organs Table 1. The mean brain weight of control animals was 1. ATP content in the cerebellum a , hippocampus b , frontal cortex c and striatum d after MDMA administration to adolescent rats. Statistical comparisons were made using t-test for the quinoprotein levels in cerebellum and striatum and protein carbonylation in cerebellum, hippocampus and frontal cortex; and Mann-Whitney Rank Sum test for quinoprotein levels in hippocampus and frontal cortex, and for protein carbonylation in striatum. Moreover, MDMA had no influence on quinoprotein levels in the brain areas. In cerebellum, hippocampus, frontal cortex and striatum no differences were found between controls and MDMA-treated animals regarding quinoprotein levels Table 2. The levels of protein carbonylation in the cerebellum, hippocampus, frontal cortex and striatum are also presented in Table 2 , and there were no significant alterations in protein carbonyl levels in the four brain areas of treated animals. There were no significant differences in these parameters between control and MDMA-treated animals. The qualitative histologic examination of peripheral organs liver, heart and kidneys of control and MDMA-treated rats was performed by means of optical microscopy. Representative histological figures can be observed in Fig. MDMA-treated rats showed sinusoidal dilatation green arrows with a marked cellular vacuolization in the periportal regions. In d scattered cardiomyocytes with signs of intracellular oedema yellow arrows can be observed, as identified by the reduced cytoplasmic staining. In f a slight interstitial oedema blue arrows , detected by the enlarged space between the tubular structures, can be observed. The control group showed a preserved liver tissue structure Fig. Livers of MDMA-treated rats presented a marked cellular vacuolization in the periportal regions, and sinusoidal dilatation with periportal and centrilobular vascular congestion Fig. No necrotic zones or interstitial inflammatory cell infiltration was observed in either group. In the histological analysis of the heart, both controls Fig. MDMA exposed animals presented random signs of cardiomyocyte oedema, particularly in the sub-endocardic region Fig. The renal tissue organization remained preserved in the control group Fig. However, the MDMA-treated group presented scattered interstitial oedema, detected by the enlarged space between the tubular structures, and signs of vascular congestion Fig. No differences were found for all these parameters in the three organs, between control and MDMA-treated animals. ATP levels were measured in the liver, heart and kidneys, and no significant differences in the ATP content were observed between control and MDMA-treated animals, as can be seen in Table 4. In Fig. No differences were found regarding this parameter in the heart or kidneys when comparing both groups Fig. Quinoprotein levels in liver a , heart b and kidneys c of control and MDMA-treated adolescent rats. In Table 4 are presented the results of protein carbonylation in the liver, heart and kidneys in the two groups. No significant differences were found in the levels of protein carbonyls in the liver and heart between groups. Data concerning the activities of caspase-3, -8 and -9 in liver, heart, and kidneys in the two groups are presented in Table 5. No differences were found in the activity of this protease either in heart or kidneys. The activities of caspase-3 and caspase-9 had no significant alterations in all three organs following MDMA when compared to the control group. Caspase-3, -8 and -9 activities in the heart, kidneys and liver of adolescent rats that received either saline or MDMA. Using a dose that does not evoke serotonergic neurotoxicity in adolescent rats we did not observe any signs of brain oxidative stress and the toxic effects occurred mainly in the peripheral organs. Animals reproduce the MDMA-induced hyperthermia seen in humans. Therefore, we proved that lower MDMA doses do produce hyperthermia in adolescent rats. MDMA abuse promotes several physiological changes, and, for that reason, we also evaluated body weight gain, as well as water and food consumption. In the literature, the cardiovascular changes and anorectic actions of amphetamines and MDMA are well described \[ 2 \]. A decrease in body weight was found in both groups of animals with similar levels, possibly a consequence of stress evoked by animal handling throughout the experiment. Our acute protocol neither evoked slower weight gain nor animal dehydration, which is seen in protocols of animal MDMA exposure during several days \[ 37 \]. Therefore, it appears that dehydration has not an important role in our overall results. Furthermore, studies in vitro in neuronal cultures revealed that the MDMA-induced neurotoxic effects are potentiated by hyperthermia \[ 26 , 39 \]. A report using adult 10 week old Wistar rats that received the same MDMA binge scheme of our study, revealed that under normal ambient temperature Therefore there is a tight relation between body temperature and MDMA serotonergic neurotoxicity \[ 16 \]. Altogether, our results show that hyperthermia per se is not the triggering factor for serotonergic toxicity or to other neurotoxic actions. Nevertheless, the hyperthermia induced by MDMA certainly potentiates the toxicity found in the brain and most importantly in the peripheral organs. Despite the absence of serotonergic neurotoxicity, we sought to study other markers of brain toxicity, namely energetic status and oxidative stress parameters. It is important to know whether doses that do not cause depletion of monoamines can elicit other toxic brain changes. Most likely there is a possible transitory effect in brain ATP levels caused by MDMA, and the time frame may disclose that short periods following exposure a decrease in ATP levels occurs, meanwhile longer periods following exposure ATP brain levels can recover. Of note that only the frontal cortex area showed a decrease in the ATP levels in the present study. The involvement of this area in both memory and decision-making is widely known, however it is unclear whether this event is related with the impairment in memory and learning abilities seen in young animals exposed to MDMA \[ 7 , 8 \]. More investigation is needed in adolescent animals to confirm the long-lasting effects of amphetamines in ATP brain levels. The metabolism of MDMA is a known triggering factor for the toxicity of this drug. The MDMA metabolites are highly reactive and can evoke oxidative stress \[ 1 \]. Several studies showed that MDMA metabolites promote neurotoxic effects to laboratory animals \[ 43 , 44 \]. Catechol MDMA metabolites were also shown to promote toxicity to cardiomyocytes \[ 17 \] and to hepatocytes \[ 47 \] in vitro. There are other contributing factors for MDMA-induced oxidative stress, including monoamine neurotransmitters metabolism by monoamine oxidase \[ 27 \], and nitric oxide formation leading to damaging reactive nitrogen species \[ 1 \]. Our paradigm of MDMA exposure elicited no oxidative stress related changes in the adolescent rat brain. Other studies reported decreases in glutathione levels \[ 48 \] and increases in protein carbonylation \[ 27 \] in the rat brain after MDMA exposure. Major differences between the previously mentioned studies and the current study are the use of higher doses, older animals, and different time-points at measurements. The lack of changes in brain oxidative stress parameters following MDMA-induced hyperthermia confirms that hyperthermia induction per se is not a guarantee for MDMA-evoked brain oxidative stress. Regarding the oxidative stress related parameters evaluated in the liver, heart, and kidneys, we could only find an increase in quinoproteins in the liver following MDMA exposure. In fact, MDMA metabolism is primarily hepatic, and, as previously mentioned, promotes the formation of catechol metabolites that can generate protein-bound quinones. Moreover, in rat hepatocytes, catechol MDMA metabolites promoted ortho-quinones formation and oxidative stress \[ 47 \]. The increase in liver quinoprotein formation possibly reflects the contribution of MDMA metabolism and the formation of reactive metabolites, and reveals the higher susceptibility of the liver to MDMA-evoked toxicity. Using higher doses and older animals, others reported decreases in glutathione levels. The fact that our low MDMA binge dose did not elicit changes in GSH levels in the peripheral organs 24 h following exposure, reveals that GSH levels might not have been affected or that GSH could decrease at early times of exposure but then recovered. Enhanced carbonylation in the kidney has been associated with the development of hypertension and kidney disease \[ 49 \]. Importantly, MDMA administration was shown to increase blood pressure in humans \[ 36 \], as well as in laboratory animals \[ 4 \]. In accordance, both the histological damage and the trend for protein carbonylation increase that we found in the kidneys reveal that this organ may be highly prone to damage following MDMA. Vascular alterations that we observed in the three studied organs, including vascular congestion, after exposure to MDMA have been associated to the MDMA-elicited hyperthermic response \[ 50 \]. In fact, other studies reported vascular lesions in the peripheral organs as a consequence of hyperthermia \[ 51 , 52 \]. The exposure of Wistar rats to high temperature environments was previously shown to result in several vascular lesions in the heart, liver, kidneys, and lungs of animals that can possibly lead to functional organ failure \[ 52 \]. Those effects were similar to the signs of damage observed in our study. Halpin and co-workers reported morphological damages in the liver of rats 24 h after the treatment with METH. The referred morphological changes were prevented when the hyperthermic response induced by the treatment with METH was blocked, suggesting that liver damage is possibly a consequence of METH-induced hyperthermia \[ 51 \]. Therefore, it can be postulated that the hyperthermic response observed in our experiment may have contributed to the observed histological alterations. The liver may be an organ with greater susceptibility to MDMA toxicity as indicated not only by the tissue damage, but also because it was the only to show an increase in quinoprotein formation. Of note, that one of the most frequently reported damage promoted by MDMA in humans is hepatotoxicity \[ 18 , 24 , 25 \]. In fact, MDMA liver metabolism and hyperthermia may cooperate to render the liver very vulnerable to damage. Moreover, the absence of caspase activity increase in the organs proves the lack of severe damage to the tissues, given that caspases are important effectors of the apoptotic pathway \[ 53 \]. The notable exception of caspase-8 activity in the liver, which revealed a significant decrease in MDMA-treated rats, might be related to a repression of genes related to apoptosis. The inhibition of caspase-8 activity has been observed in hepatic cells through nitric oxide signalling \[ 54 \]. In cultured rat striated cardiac myocytes there was a repression of caspase-1 and caspase-8 genes following exposure to MDMA \[ 55 \]. The heart is particularly susceptible to oxidative stress-related injuries and amphetamines like MDMA evoke cardiotoxicity \[ 56 \]. Our study used lower MDMA doses in adolescent rats and drug exposed hearts showed a particular vulnerability of the myocytes from the sub-endocardic region. Reports from MDMA users following fatalities describe major organ changes, including necrosis, oedema and inflammation \[ 1 , 24 , 25 \]. These important human findings report a rather extreme scenario following the course of MDMA intoxication. Our report more reliably reproduces the hyperthermia seen in human abusers and organ changes might be more similar to those seen in adolescents. MDMA moderate doses, more close to those used by human adolescents, do not elicit in adolescent rats oxidative stress related changes in the brain. MDMA treatment in adolescent rats promoted morphological tissue alterations in the heart, kidneys, and liver, as well as rises in liver quinoproteins. New studies are required to assess the impact of moderate MDMA doses in adolescent animals to verify whether these brain and peripheral organ changes are long-lasting and may be reflected later during adulthood. ATG, VMC and JPC have made substantial contributions to conception and design of the study, were involved in all experimental procedures and drafted the manuscript. JAD performed the experimental procedures and data analysis of organ histology. JPC conceived of the study. All authors contributed to the interpretation of data and manuscript writing. All authors read, revised and approved the final manuscript. Additionally, part of the work has been present orally at the Portuguese Pharmacology Society annual meeting, and in a Poster at the Eurotox international meeting. This section collects any data citations, data availability statements, or supplementary materials included in this article. As a library, NLM provides access to scientific literature. BMC Pharmacol Toxicol. Find articles by Armanda Teixeira-Gomes. Find articles by Vera Marisa Costa. Find articles by Rita Feio-Azevedo. Find articles by Eduarda Fernandes. Find articles by Maria de Lourdes Bastos. Received Nov 11; Accepted Jun 3; Collection date Open in a new tab. Effect of MDMA administration in oxidative stress related parameters in the four brain areas. Effect of MDMA administration in caspase-3, -9 and -8 activities in the three peripheral organs. Similar articles. Add to Collections. Create a new collection. Add to an existing collection. Choose a collection Unable to load your collection due to an error Please try again. Add Cancel.

Buy Ecstasy Leuven