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Marijuana Party of Canada-CANCER CURE- Candidate

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#1 bigdaddyrolling7



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Posted 26 December 2007 - 10:43 AM

Marijuana Party of Canada-CANCER CURE- Candidate Rick SimpsonGlobal
Group Info Name: Marijuana Party of Canada-CANCER CURE- Candidate Rick Simpson
Type: Common Interest - Politics
Rick Simpson will be running in the next Federal Election
© 2003-2007 Parti Marijuana // Marijuana Party
a Party Representive for Cumberland, Colchester and Musquodoboit Valley

This is the very first Marijuana Party Candidate this riding ever had "THIS IS HISTORY IN THE MAKING"

Lets help send the MAN with the "CURE FOR CANCER" to Ottawa when the election is called 20,000-25,000 votes would send him.

1.5 million Canadians have a record for hemp and WHY?
at the rate of60- 70,000 cannabis cannadians charges a year for HEMP?
The government is promising more crack downs on our
Culture and Manditory Jail,all for What,and Why?

Why should our Children have Records,and go to Jail because of Hemp?

Hemp is mans safest and oldest medicine and was known as a PANACEA-CURE ALL and when made into THC Hemp Oil thats what you got.

Not one death and the scientific studies to prove it
hemp has over 25,000 uses THC KILLS CANCER CELLS[FACT]

8 Canadians die an hour from cancer and 1 in 2 will get a form of cancer

vote for the Marjuana party and vote for Rick Simpson
he has been curing cancer since 2003 for free

Coming soon TO- DVD EVERY- WHERE



if you read just one of the post here
read grinspoon ......and the recent news at the bottom of this page.....
Become a Registered Member
Become a Registered Member
Free Memberships to Elections Canada.
Sunday July 1st, 2007 | M | français

The “Marijuana Party” IS
an association of electors.

Elections law requires us
to prove we have 250,
every three years ...

Marijuana Party members,
complete and mail us one

“Membership Declaration”

Elections Canada form: EC 20036-C 02/07

You may obtain a copy of this

Membership Declaration

by downloading & printing the PDF file
from button at bottom of this Web page.

(You could also mail us a request,
and we would send you the form.)

Alex Neron, Marijuana Party’s 1st officially registered member.

The Marijuana Party encourages interested electors to
obtain declarations, fill them in, sign, & mail them to:

Parti Marijuana Party
23 - 3865 rue Bélanger
Montréal, QC H1X 1B4

Or contact us

Party leader forwards declarations to
the Chief Electoral Officer, as required.

Membership in the Marijuana Party is free.

Every three years, a registered party MUST
submit 250 Membership Declarations
signed during that year ...

Between January 1, 2010, and June 30, 2010,
the Marijuana Party must again get 250 plus
adult citizens’ membership declarations, to
submit to Elections Canada to continue to
meet the requirements of the elections law ...

(A party failing to do so
may get extensions from
Elections Canada to the
end of 2010, however not
submitting 250 by 2011
forces de-registration.)

Membership Declarations submitted from 2007 to 2009
will be used in the future to generate a mailing list
sent requests to sign the triannual Declarations
as required in 2010, 2013, 2016, and so on ...

A signing member must be a "Canadian Elector,"
and write their residential address as an elector.

Elections Canada only accepts memberships
that have their complete residential address,
which is the address that elector who signs
should have as an elector on a voting list.

Merely stating a Post Office Box number is NOT sufficient.

We would like to also have your mailing address, but
you must write your complete residential address,
which is your home address as Canadian Elector,
in order for that to be counted and registered.

The Marijuana Party is governed
solely by Canada Elections Law.

Signing member declarations
is free, and does not oblige
members to future actions.

Signing an Elections Canada

Membership Declaration

ONLY fulfills the requirements in
current Canada elections law.

Nothing else follows from
signing and submitting
those declarations.

Signing memberships does not
require members to perform
any duties for the party.

Marijuana Party membership
may gain privileges to participipate
in some Electoral District Associations.


Elections Canada Membership Declaration EC 20036-C
(PDF, 46.5 kb)

Elections Canada Web site

Contact en | fr

Articles published in this section:

Membership Laws

© 2003-2007 Parti Marijuana // Marijuana Party

Contact Info Email: bigdaddyrollingstone@hotmail.com
Website: http://www.phoenixtears.ca
Office: Marijuana Party Candidate- Rick Simpson
Street: 344 little forks rd

Recent Newsedit

The therapeutic properties of the hemp plant, Cannabis sativa, have been known since antiquity, but the recreational use of its euphoric and other psychoactive effects has restricted for a long time research on its possible pharmaceutical application. The isolation of 9-tetrahydrocannabinol (THC), the main psychoactive component of Cannabis (Gaoni and Mechoulam, 1964), opened the way to further investigations. After the discovery of the two specific molecular targets for THC, CB1, and CB2 (for review, see Pertwee, 1997), it became clear that most of the effects of marijuana in the brain and peripheral tissues were due to activation of these two G-protein-coupled cannabinoid receptors. However, evidence is also accumulating that some pharmacological effects of marijuana are due to Cannabis components different from THC. Indeed, C. sativa contains at least 400 chemical components, of which 66 have been identified to belong to the class of the cannabinoids (Pertwee, 1997).

To date, cannabinoids have been successfully used in the treatment of nausea and vomiting (for review, see Robson, 2005), two common side effects that accompany chemotherapy in cancer patients. Nevertheless, the use of cannabinoids in oncology might be somehow underestimated since increasing evidence exist that plant, synthetic, and endogenous cannabinoids (endocannabinoids) are able to exert a growth-inhibitory action on various cancer cell types. However, the precise pathways through which these molecules produce an antitumor effect has not been yet fully characterized, also because their mechanism of action appears to be dependent on the type of tumor cell under study. It has been reported that cannabinoids can act through different cellular mechanisms, e.g., by inducing apoptosis, cell-cycle arrest, or cell growth inhibition, but also by targeting angiogenesis and cell migration (for review, see Bifulco and Di Marzo, 2002; Guzman, 2003; Kogan, 2005). Furthermore, the antitumoral effects of plant, synthetic and endocannabinoids can be mediated by activation of either CB1 (Melck et al., 2000; Bifulco et al., 2001; Ligresti et al., 2003; Mimeault et al., 2003) or CB2 receptors or both (Sanchez et al., 2001; Casanova et al., 2003; McKallip et al., 2005), and, at least in the case of the endocannabinoid anandamide, by transient receptor potential vanilloid type-1 (TRPV1) receptors (Maccarrone et al., 2000; Jacobsson et al., 2001; Contassot et al., 2004) as well as by noncannabinoid, nonvanilloid receptors (Ruiz et al., 1999). Additionally, cannabidiol has been suggested to inhibit glioma cell growth in vitro and in vivo independently from cannabinoid and vanilloid receptors (Massi et al., 2004; Vaccani et al., 2005).

The main limitation of the possible future use of THC in oncology might be represented by adverse effects principally at the level of the central nervous system, consisting mostly of perceptual abnormalities, occasionally hallucinations, dysphoria, abnormal thinking, depersonalization, and somnolence (Walsh et al., 2003). However, most non-THC plant cannabinoids seem to be devoid of direct psychotropic proprieties. In particular, it has been ascertained that cannabidiol is nonpsychotropic (for review, see Mechoulam et al., 2002; Pertwee, 2004) and may even mitigate THC psychoactivity by blocking its conversion to the more psychoactive 11-hydroxy-THC (Bornheim and Grillo, 1998; Russo and Guy, 2006). Moreover, it has been recently found that systematic variations in its constituents (i.e., cannabidiol and cannabichromene) do not affect the behavioral or neurophysiological responses to marijuana (Ilan et al., 2005). Finally, it has been also shown that, unlike THC, systemic administration to rats of cannabigerol does not provoke poly-spike discharges in the cortical electroencephalogram during wakefulness and behavioral depression (Colasanti, 1990). These and other observations reinforce the concept that at least cannabidiol, cannabigerol, and cannabichromene lack psychotropic activity and indicate that for a promising medical profile in cancer therapy, research should focus on these compounds, which instead have been poorly studied with regard to their potential antitumor effects. By keeping this goal in mind, we decided to investigate the antitumor properties of cannabigerol and cannabichromene. We also screened THC acid and cannabidiol acid and two distinct Cannabis extracts (enriched in either cannabidiol or THC), where the presence of nonpsychotropic cannabinoids along with THC has been reported to mitigate the potential side effects of the latter compound in clinical trials (Russo and Guy, 2006).

All plant cannabinoids, the two cannabinoid acids, and the two Cannabis extracts were kindly provided by GW Pharmaceuticals. Cannabidiol- and THC-rich extracts contained approximately 70% cannabidiol or THC, respectively, together with lesser amounts of other cannabinoids. The two cannabinoid receptor antagonists, SR141716A and SR144528, were a kind gift from Sanofi-Aventis (Paris, France), whereas methyl--cyclodextrin, all of the antioxidant drugs (-tocopherol, vitamin C, astaxantine), N-Acetyl-Asp-Glu-Val-Asp-aldehyde, and BAPTA-AM were purchased from Sigma-Aldrich (St. Louis, MO). The endocannabinoid uptake inhibitor (S)-1'-(4-hydroxybenzyl)-N-ethyl-oleoylamide was synthesized as previously described in Ortar et al. (2003). Finally, all of the TRPV1 or cannabinoid receptor agonists and antagonists (capsaicin, resiniferatoxin, arachidonoyl-2-chloro-ethylamide, JWH-133, AM251, AM630) were obtained from Tocris Cookson (Bristol, UK).

Cell lines from various origins (MCF-7 and MDA-MB-231 human breast carcinoma cells, DU-145 human prostate carcinoma cells, CaCo-2 human colorectal carcinoma cells, AGS human gastric adenocarcinoma cells, C6 rat glioma cells, KiMol rat thyroid cells transformed with the v-K-ras oncogene, and rat basophilic leukemia cells) were maintained at 37°C in a humidified atmosphere containing 5% CO2. Media, sera, and subculturing procedures differed from line to line and were according to the information provided in each case by the supplier company (DSMZ, Braunschweig, Germany). Primary cells derived from normal human mammary glands were purchased from Cell Applications, Inc. (San Diego, CA) and cultured as described in the data sheet from the supplier.

Cell Proliferation Assay. Six-well culture plates were incubated at 37°C at a cell density of 5 x 104 cells/well in a humidified atmosphere containing 5% CO2. Three hours after seeding, vehicle or cannabinoids at different concentrations were added to the medium and then daily with each change of medium for 4 days, and the effect of compounds on cell growth was measured by Crystal Violet vital staining. After staining, cells were lysated in 0.01% acetic acid and analyzed by spectrophotometer analysis (PerkinElmer Lambda 12, = 595 nm; PerkinElmer Life and Analytical Sciences, Boston, MA). Optical density values from vehicle-treated cells were considered as 100% of proliferation. Statistical analysis was performed using ANOVA followed by Bonferroni test.

Detection of Reactive Oxygen Species. Intracellular reactive oxygen species (ROS) generation was determined by spectrofluorometric analysis. MDA-MB-231 cells were plated (16 x 103 cells/well) in Porvair PS-White Microplate 96-well (PerkinElmer Life and Analytical Sciences) for 12 h. The day of the experiment, cells were rinsed once with Tyrode's buffer, then loaded (1 h at 37°C in darkness) with 10 µM 2',7'-dichlorofluorescein diacetate (fluorescent probe; Molecular Probes, Eugene, OR) in the presence of 0.05% Pluronic F-127. Reactive oxygen species (ROS)-induced fluorescence of intracellular
2',7'-dichlorofluorescein diacetate was measured with a microplate reader (PerkinElmer LS50B, Ex, 495 nm; Em, 521 nm). Fluorescence detections were carried out after the incubation of 100 µM H2O2 and/or increasing concentrations of cannabidiol at room temperature in the darkness for different times (0-30-60-120 min). The fluorescence measured at time 0 was considered as basal ROS production and subtracted from the fluorescence at different times (1). Data are reported as mean ± S.E. of 2, i.e., fluorescence 1 values at different doses subtracted of the 1 values of cells incubated with vehicle. In some experiments, a buffer containing MgCl2 in amounts equivalent to CaCl2 and 0.1 mM EGTA and cells preloaded for 30 min with BAPTA-AM (40 µM) were used instead.

Reverse Transcription-Polymerase Chain Reaction Analysis. Total RNAs from these cells were extracted using the Trizol reagent according to the manufacturer's recommendations (Invitrogen, Carlsbad, CA). Following extraction, RNA was precipitated using ice-cold isopropanol, resuspended in diethyl pyrocarbonate (Sigma-Aldrich)-treated water, and its integrity was verified following separation by electrophoresis on a 1% agarose gel containing ethidium bromide. RNA was further treated with RNase-free DNase I (Ambion DNA-free kit; Ambion, Austin, TX) according to the manufacturer's recommendations to digest contaminating genomic DNA and to subsequently remove the DNase and divalent cations.

The expression of mRNAs for CB1, CB2, TRPV1, and GAPDH were examined by semiquantitative RT-PCR. Total RNA was reverse-transcribed using random primers. DNA amplifications were carried out in PCR buffer (Invitrogen) containing 2 µl of cDNA, 500 µM dNTP, 2 mM MgCl2, 0.8 µM of each primer, and 0.5 U of Taq polymerase platinum (Invitrogen). The thermal reaction profile consisted of a denaturation step at 94°C for 1 min, annealing at 55°C (GAPDH) or 57°C (CB2 and TRPV1) or 60°C (CB1) for 1 min, and an extension step at 72°C for 1 min. A final extension step of 10 min was carried out at 72°C. The PCR cycles observed to be optimal and in the linear portion of the amplification curve were 24 for GAPDH, 29 for CB1 and CB2, and 28 for TRPV1 (data not shown). Reaction was performed in a PE Gene Amp PCR System 9600 (PerkinElmer Life and Analytical Sciences). After reaction, the PCR products were electrophoresed on a 2% agarose gel containing ethidium bromide for UV visualization.

Specific rat and human oligonucleotides were synthesized on the basis of cloned rat and human cDNA sequences of CB1 (GenBank accession nos. NM_012784 [GenBank] .3 and X81120 [GenBank] for rat and human, respectively), CB2 (GenBank accession nos. NM_0205433 and X74328 [GenBank] for rat and human, respectively), TRPV1 (GenBank accession nos. NM_031982 [GenBank] and NM_080706 [GenBank] .2 for rat and human, respectively), and GAPDH (GenBank accession nos. NM_017008 [GenBank] .2 and BT006893 [GenBank] .1 for rat and human, respectively).

For rat and human CB1, the primers sequences were 5'-GAT GTC TTT GGG AAG ATG AAC AAG C-3' (nt 1250-1274 for rat and nt 1187-1211 for human; sense) and 5'-AGA CGT GTC TGT GGA CAC AGA CAT GG-3' (nt 1558-1534 for rat and nt 1495-1470 for human; antisense). The rat CB2 sense and antisense primers were 5'-TA(C/T) CC(G/A) CCT (A/T)CC TAC AAA GCT C-3' (nt 407-428) and 5'-C (A/T)GG CAC CTG CCT GTC CTG GTG-3' (nt 698-676), respectively. For human CB2, the primers sequences were 5'-TTT CCC ACT GAT CCC CAA TG-3' (nt 672-691; sense) and 5'-AGT TGA TGA GGC ACA GCA-3' (nt 1000-983; antisense). For rat TRPV1, the primers sequences were 5'-GAC ATG CCA CCC AGC AGG-3' (nt 2491-2508; sense) and 5'-TCA ATT CCC ACA CAC CTC CC-3' (nt 2752-2733; antisense). The human TRPV1 sense and antisense primers were 5'-TGG ACG AGG TGA ACT GGA C-3' (nt 2761-2779) and 5'-ACT CTT GAA GAC CTC AGC GTC-3' (nt 3023-3003), respectively. For rat and human GAPDH, the primers sequences were 5'-CCC TTC ATT GAC CTC AAC TAC ATG GT-3' (nt 949-974 for rat and nt 106-131 for human; sense) and 5'-GAG GGG CCA TCC ACA GTC TTC TG-3' (nt 1418-1396 for rat and nt 575-553 for human; antisense).

The expected sizes of the amplicons were 309 bp for rat and human CB1, 291 bp for rat CB2, 329 bp for human CB2, 263 bp for rat TRPV1, 262 bp for human TRPV1, and 470 bp for rat and human GAPDH. In the presence of contaminant genomic DNA, the expected size of the amplicons would be 1062 bp for GAPDH (data not shown). No PCR product was detected when the reverse transcriptase step was omitted.

Western Immunoblotting Analysis for Caspase-3. Immunoblotting analysis was performed on the cytosolic fraction of cells treated as described above and according to previous published work (Iuvone et al., 2004). Cytosolic fraction proteins were mixed with gel loading buffer (50 mM Tris/10% SDS/10% glycerol 2-mercaptoethanol/2 mg of bromphenol/ml) in a ratio of 1:1, boiled for 5 min and centrifuged at 10,000g for 10 min. Protein concentration was determined, and equivalent amounts (50 µg) of each sample were separated under reducing conditions in 12% SDS-polyacrylamide minigel. The proteins were transferred onto nitrocellulose membrane, according to the manufacturer's instructions (Bio-Rad, Hercules, CA). The membranes were blocked by incubation at 4°C overnight in high-salt buffer (50 mM Trizma base, 500 mM NaCl, 0.05% Tween 20) containing 5% bovine serum albumin and then incubated for 2 h with anticaspase 3 (1:2000, v/v) at room temperature, followed by incubation for 2 h with horseradish peroxidase-conjugate secondary antibody (Dako, Glostrup, Denmark). The immune complexes were developed using enhanced chemiluminescence detection reagents (GE Healthcare, Little Chalfont, Buckinghamshire, UK) according to the manufacturer's instructions and exposed to Kodak X-OMAT film (Eastman Kodak, Rochester, NY). The bands of protein on X-ray film were scanned and densitometrically analyzed with a GS-700 imaging densitometer.

Immunofluorescence. For immunoreaction, MDA-MB-231 cells were seeded on sterile coverslips (22 x 22 mm; Menzel, Braunschweig, Germany) in six-well culture plates and incubated under standard conditions until they were at least 70% confluent. Cultured cells were processed for immunofluorescence. After three washes with PBS, cells were fixed by incubating them in 4% (v/v) paraformaldehyde in PBS for 20 min at room temperature, rinsed with PBS, permeabilized for 15 min in 0.5% Triton X-100 in PBS, and incubated overnight at 4°C with rabbit polyclonal rabbit anti-CB1 or anti-CB2 antibody (Cayman Chemical, Ann Arbor, MI), both diluted 1:50 in 0.5% Triton X-100 in PBS, or goat anti-TRPV1 antibody (Santa Cruz Biochemicals, Santa Cruz, CA) diluted 1:100 in 0.5% Triton X-100 in PBS. After three washes in PBS, fluorescence was revealed by incubation for 2 h in an Alexa Fluor 488-labeled secondary anti-rabbit antibody (Invitrogen) diluted 1:100 in 0.5% Triton X-100 in PBS or Alexa Fluor 546-labeled secondary anti-goat antibody (Invitrogen) diluted 1:200 in 0.5% Triton X-100 in PBS. The preabsorption of antibodies with the respective blocking peptides as well as omission of primary antibodies (control immunoreaction) resulted in much weaker or negative staining, respectively. Sections processed for immunofluorescence were studied with an epifluorescence microscope equipped with the appropriate filter (Leica DM IRB; Leica, Wetzlar, Germany). Images were acquired using a digital Leica DFC 320 camera connected to the microscope and the image analysis software Leica IM500. Images were processed in Adobe Photoshop (Adobe Systems, Mountain View, CA), with brightness and contrast being the only adjustments made.

In Vivo Studies: Effect on Xenograft Models of Carcinoma. All of the experiments were performed by using Charles River 6-week-old male athymic mice (Charles River, Margate, Kent, UK) as described previously (Bifulco et al., 2001). Two different mouse xenograft models of tumor growth were induced by s.c. injection (5 x 105 cells) of two distinct highly invasive tumoral cell lines (KiMol or MBA-MD-231 cells) into the dorsal right side of athymic mice. Starting from the appearance of tumoral mass, pure compounds or Cannabis extracts were injected intratumor in the same inoculation region twice per week for 20 (KiMol cell-induced tumors) or 16 (MBA-MD-231 cell-induced tumors) days. THC and cannabidiol were administered at the dose of 5 mg/kg, whereas THC-rich and cannabidiol-rich were administrated at the dose of 6.5 mg/kg, which contains 5 mg/kg THC and cannabidiol, respectively. Tumor diameters were measured with calipers every other day until the animals were killed. Tumor volumes (V) were calculated by the formula of rotational ellipsoid (V = AxB2/2; A = axial diameter, B = rotational diameter). Results were reported as means ± S.E. Statistical analysis was performed using ANOVA followed by the Bonferroni's test.

In Vivo Analysis: Effect on Experimental Lung Metastasis. Monocellular suspension of MDA-MB-231 cells containing 2.5 x 105 cells was injected into the left paw of 30-day-old BalB/c male mice. Animals were divided into three groups: vehicle (n = 11), cannabidiol (5 mg/kg/dose, n = 14), or cannabidiol-rich (6.5 mg/kg/dose, n = 14). The drugs were injected i.p. every 72 h. Experimental metastases were evaluated 21 days after the injection. To contrast lung nodules, lungs were fixed in Bouin's fluid, and metastatic nodes were scored on dissected lung under a stereoscopic microscope. All animal studies were conducted in accordance with the Italian regulation for the welfare of animals in experimental neoplasia. All data were presented as means ± S.D. Statistical analysis was performed using one-way ANOVA.

Cell Cycle and Apoptosis Detection. Different cell lines were exposed to 10 µM of cannabidiol or cannabigerol for 48 h at 37°C in a humidified atmosphere containing 5% CO2. The distribution of cells among the different phases of the cell cycle and apoptosis rate were evaluated by flow cytometric analysis of the DNA content. Cells (5 x 105) were collected, washed twice with PBS, fixed by ethanol 70%, and kept at -20°C for at least 4 h. Propidium iodide (10 µg/ml) in PBS containing 100 U/ml DNase-free RNase was added to the cells for 15 min at room temperature. Cells were acquired by a FACScalibur flow cytometer (BD Biosciences, San Jose, CA), and then analysis was performed using ModFit LT version 3.0 from Verity Software House, Inc. (Topsham, ME); 10,000 events were collected and corrected for debris and aggregate populations.

Anandamide Cellular Reuptake and Intracellular Hydrolysis. The effect of compounds on anandamide cellular reuptake was analyzed on rat basophilic leukemia cells or MDA-MB-231 cells by using 2.5 µM (10,000 cpm) [14C]anandamide as described previously (De Petrocellis et al., 2000). Briefly, cells were incubated with [14C]anandamide for 5 min at 37°C, in the presence or absence of varying concentrations of the compounds. Residual [14C]anandamide in the incubation medium after extraction with CHCl3/CH3OH 2:1 (by volume), determined by scintillation counting of the lyophilized organic phase, was used as a measure of the anandamide that was taken up by cells (De Petrocellis et al., 2000). Nonspecific binding of [14C]anandamide to cells and plastic dishes was determined in the presence of 100 µM anandamide and was never higher than 30%. Data are expressed as the concentration exerting 50% inhibition of anandamide uptake (IC50) calculated by GraphPad software (GraphPad Software Inc., San Diego, CA). The effect of compounds on the enzymatic hydrolysis of anandamide was studied using membranes prepared from N18TG2 cells, incubated with the test compounds and [14C]anandamide (20,000 cpm; 5 µM) in 50 mM Tris-HCl, pH 9, for 30 min at 37°C. [14C]Ethanolamine produced from [14C]anandamide hydrolysis was measured by scintillation counting of the aqueous phase after extraction of the incubation mixture with 2 volumes of CHCl3/CH3OH 1:1 (by volume). Data are expressed as the concentration exerting 50% inhibition of [14C]anandamide hydrolysis (IC50), calculated by GraphPad software.

Activity at Human Recombinant TRPV1. The effect of the substances on [Ca2+]i was determined by using Fluo-3 (Molecular Probes), a selective intracellular fluorescent probe for Ca2+ (De Petrocellis et al., 2000). Human embryonic kidney (HEK) 293 cells stably overexpressing human TRPV1 receptor or MDA-MB-231 cells were transferred into six-well dishes coated with poly-L-lysine (Sigma-Aldrich) 1 day prior to experiments and grown in the culture medium mentioned above. On the day of the experiment, the cells (50-60,000 cells/well) were loaded for 2 h at 25°C with 4 µM Flu-3-methylester (Invitrogen) in dimethyl sulfoxide containing 0.04% Pluronic F-127 (Invitrogen). After loading, cells were washed with Tyrode's solution, pH 7.4, trypsinized, resuspended in Tyrode's solution, and transferred to the cuvette of the fluorescence detector (PerkinElmer LS50B) under continuous stirring. Experiments were carried out by measuring cell fluorescence at 25°C (EX = 488 nm, EM = 540 nm) before and after the addition of the test compounds at various concentrations. Data are expressed as the concentration exerting a half-maximal effect (EC50). The efficacy of the effect was determined by comparing it with the analogous effect observed with 4 µM ionomycin. In some experiments with MDA-MB-231 cells, the effect of cannabidiol was measured also in the absence of extracellular Ca2+ (i.e., in a Tyrode's solution containing Mg2+ instead of Ca2+ and 0.1 mM EGTA) and in cells preloaded with BAPTA-AM (20 µM).

Effect on Cancer Cell Growth: In Vitro Studies. For in vitro studies, the cannabinoids under investigation were screened for their ability to reduce cell proliferation on a collection of tumoral cell lines. Cannabidiol always exhibited the highest potency with IC50 values ranging between 6.0 ± 3.0 and 10.6 ± 1.8 µM. Cannabidiol acid was the least potent compound. Among the other plant cannabinoids, cannabigerol was almost always the second most potent compound, followed by cannabichromene. The effect of the two Cannabis extracts (enriched in cannabidiol or THC) was next investigated, and in some circumstances, the cannabidiol-rich extract appeared slightly more potent than pure cannabidiol. In the case of MCF-7 cells, both compounds exhibited quite similar potency, as indicated by the IC50 values of 8.2 ± 0.3 and 6.0 ± 1.0 µM, respectively, for cannabidiol and cannabidiol-rich extract, on the contrary, in the case of C6 glioma cells, cannabidiol-rich extract also exhibited significantly higher potency than pure cannabidiol (IC50, 4.7 ± 0.6 and 8.5 ± 0.8 µM, respectively, p < 0.05, Fig. 2B). Only in the case of human DU-145 prostate carcinoma cells, plant cannabinoids induced a stimulatory effect on cancer growth at the lowest doses tested and an inhibitory effect only at the highest concentration tested (25 µM) (as also found by Sanchez et al., 2003 in another prostate carcinoma cell line). In this case, however, the cannabidiolrich extract lacked the pro-proliferative effect even at the lowest concentration tested of 2 µM

The Effect of cannabinoids and Cannabis extracts on cancer cell growth Various epithelial cell lines of various tumoral origin were treated with different concentrations of drugs, and after 4 days, the cell number was measured with Crystal Violet Vital staining (see Materials and Methods). Data are reported as mean ± S.E. of IC50 values (micromolar) calculated from three independent experiments. CBG, cannabigerol; CBC, cannabichromene; CBD-A, cannabidiol-acid; THC-A, THC-acid; CBD-rich, cannabidiol-enriched cannabis extract; THC-rich, THC-enriched cannabis extract.

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Legalize pot smoking, senators say - Pot and Politics: Canada and the Marijuana Debate - CBC Archive
Should pot be sold at corner stores to anyone over 16? Some of Canada's most senior politicians think so. The Senate Special Committee on Illegal Drugs final report says marijuana is less harmful than alcohol and should be governed by the same sort of rules. ...
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Cure Cancer with Hemp Oil - Phoenix Tears
Phoenix Tears is a not for profit entity dedicated to the production of Hemp medicines and providing information about the use of natural Hemp oil, (not Hemp Seed oil) as an effective treatment for cancer and other serious illnesses.
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