The obesity epidemic is a major health threat affecting one in four people in the affluent western world, where high-energy foods are easily available and there is little need for exercise. To identify novel therapeutic targets for the treatment of obesity, one important step is to further define the complex circuitry in the brainwhich is ultimately responsible for our appetite and body weight regulation. Although complex, appetite can be thought of as having two distinct, though none mutually exclusive, aspects: the need to eat (homeostatic) and the desire to eat(hedonistic).The need to eat, a product of energy homeostasis, is what drives the consumption offood for basic survival. In an attempt to further define the mainly “homeostatic” neuronal circuitry, we combined blood-oxygen-level-dependent (BOLD)pharmacological-challenge magnetic resonance imaging (phMRI) with c-Fosfunctional activity mapping to characterise “whole brain” responsiveness to anorexigenic dose of the glucose anti metabolite 2-deoxy-D-glucose (2-DG). Using thesecomplementary methods, we demonstrated functional brain activity in a number ofknown glucose-sensing brain regions, including parts of the hypothalamus andbrainstem, following administration of 2-DG when compared with vehicle treatment.The desire to eat is a result of a complex interplay between the reward andmotivational circuits implicated in addictive behaviours, and those which controlenergy homeostasis. Recent research has pointed to the endocannabinoid system,and specifically the central cannabinoid 1 (CB1) receptor, as a key target mediatingthe functional cross talk between the two appetitive systems. To define the sites ofaction of cannabinoids, we used an orexigenic dose of the full CB1 agonist, CP55940,to map responsive brain regions again using BOLD phMRI and whole-brain c-Fosfunctional activity mapping. Areas of interest demonstrated a drug interaction whenthe CB1 receptor inverse agonist, Rimonabant was co-administered. These complementary methods demonstrated functional activity in the cortico-striatalhypothalamicpathway, a key system in the motivational drive to eat.The appetitive actions of synthetic CB1 inverse agonists such as Rimonabant are welldocumented. We, however, described a putative novel endogenous CB1 inverseagonist, hemopressin, which is the first identified peptide ligand of CB1 receptors.We showed that hemopressin inhibits agonist-induced receptor internalisation in aheterologous cell model in vitro. When administered centrally or systemically in vivo,we found that hemopressin decreases nocturnal food intake in out-bred rats andmice, as well as in obese, leptin-deficient ob/obmice. Importantly, hemopressininduces hypophagia without causing any apparent adverse side effects. We have also shown that the anorectic effect is absent in CB1-/- mice, and that hemopressin canblock CB1 agonist-induced hyperphagia in male rats, providing strong evidence forantagonism of the CB1 receptor in vivo. We speculate that hemopressin may be one of a family of endogenous functional CB1 receptor ligands that modulate the activity of appetite pathways in the brain.

Economical analysis of market with cannabis products is solved by the master thesis. Emphasis is placed on the situation and specifics of Czech Republic market with focus on price of cannabis drugs. It contains research questions which deal with role of price of drugs users -- if price factor is more significant than other socio-economical factors. The thesis describes development of legislation and anti-drug policy in the Czech Republic and its focus on cannabis products. In the theoretical part of the thesis there are presented economical and social aspects of anti-drug policy and its forms of applications in the Czech Republic. Analytical part of the thesis is focused on analysis of recent situation in the market with cannabis products. There is presented survey dedicated to consumption of cannabis drugs by students of high (secondary) schools. The survey also deals with attitudes of these students towards cannabis drugs. The survey is the source of data for regression analysis. Research has shown the most of users of marijuana do not pay for the drug. In case of act of buying there are frequent volume distcounts. Procurations of anti-drug policy which are trying to influence the price therefore may not have any impacts on buying behavior.

Cannabinoids and cannabinoid receptors have been shown to play important roles in immune regulation particularly as modulators of anti-inflammatory cytokines and antibody production. The predominant cannabinoid receptor involved in this immune regulation is cannabinoid receptor 2 (CB2), which is robustly expressed in B cells. Utilizing a combination of bioinformatics, 5' RACE, real time RT-qPCR, and reporter assays, we showed that human B cells from peripheral blood mononuclear cells (PBMC) expressed one CB2 transcript while mouse B cells from spleen express three CB2 transcripts. Alignment of the sequenced B cell RACE products to either the mouse or human genome, along with the GenBank mRNA sequences, revealed that the transcripts isolated in this study contained previously unidentified transcriptional start sites (TSSs). In addition, expression construct testing of the genomic region containing the TSSs of the mouse CB2 exon 1 and 2 transcripts showed a significant increase of promoter activity. Bioinformatics analysis for cis-sequences in the promoter regions identified DNA binding sites for NF-kB, STAT6, and Elk1 transcription factors activated by LPS, IL-4 and anti-CD40. Regarding variations in CB2 transcript expression among the immune cell subtypes, RACE analysis showed that the exon 1b transcript is seen in B cells but not in T cells, dendritic cells or macrophages. Furthermore, RT-qPCR showed variations in transcript expression during B cell development as well as in resting versus LPS or IL-4/anti-CD40 stimulated B cells. The exon 1a transcript was predominant in pre-, immature and resting B cells whereas the exon 1b and 2 transcripts were enhanced in mature and activated B cells. These data showed for the first time that human B cells use one TSS for CB2 expression while mouse B cells use multiple TSSs for the expression of three CB2 transcripts, in which the expression of the individual transcript is related to immune cell type and/or cell activation state. Additionally, this is the first report in mouse B cells defining TSSs that are in genomic areas with promoter activity thus suggesting the location of two promoter regions. Defining the CB2 transcript expression during various stages of B cell activation provide clues to therapeutic methods.

The object of this thesis is a traditional and also controversial plant Cannabis sativa. The theoretical part is focused on ubiquitous polyphenolic compounds – flavonoids, also present in the studied plant. All previously researched and confirmed effects on human body are described in detail. The experimental part of this thesis describes the isolation and identification of substances of the chloroform and hexane yields of extract obtained from this plant. Chromatographic methods were used for isolation – thin layer and column chromatography and semipreparative HPLC. Obtained substances were characterized using UV, IR spectrophotometry and NMR. Then the anti-inflammatory activity of obtained substances and also of the ethanolic extract and its yields was observed. The result of the experimental part was the isolation of one pure substance which was identified as the cannabidiolic acid. Using the THP 1 assay, we demonstrated mild anti-inflammatory effect of the non-polar yields and isolated CBDA.

Cannabis (and its main psychoactive component, THC) is one of the most widely-used drugs in the world, and recent expansion of its legal status has made it available in a variety of formulations and at a potency unrivaled in history. While its medicinal properties are gaining scientific support, so too is its potential to lead to abuse and dependence. Both initiation of cannabis use and frequent cannabis use are most prevalent in adolescence, and compared to adults, cannabis use by adolescents is associated with a greater likelihood of developing cannabis dependence and cannabis use disorder. Given the ethical limitations surrounding research that provides cannabis to non-users or non-adults, animal models of drug use can be valuable tools for the study of causes and consequences related to drug use, as well as allowing for investigating brain mechanisms underlying these factors. However, only recently have models in which animals reliably use cannabis (THC) at levels above its respective vehicle and at levels which produce consistent behavioral and physiological effects become available, and in no case has age-related differences in this use been examined. Thus, one goal of the current study was to directly compare the self-administration of edible THC (a route of administration used by humans and a formulation increasing in popularity) between adolescent and adult mice.<br><div> Adolescents also appear to be differentially sensitive to various effects of several classes of drugs, and they have been shown to be less sensitive to the aversive effects of cannabis, thereby putting them at greater risk for elevated and continued use. Evidence also suggests that, in addition to the risk associated with adolescent cannabis use, having initial positive subjective experiences resulting from its use is a strong predictor of subsequent cannabis dependence. Thus, the second goal of the current study was to use the place conditioning paradigm to examine the reward- (or aversion-) inducing properties of THC in adolescent and adult C57BL/6J mice, using both the traditional experimenter-administered THC (via injection) as well as edible THC self-administration.</div><div> Prior to initiating these THC studies, sensitivity of the place conditioning procedure to age-related differences in drug-induced reward was validated using cocaine, yielding locomotor stimulation in both ages and a decreased sensitivity to cocaine’s rewarding properties in adolescent mice. When provided limited access to edible THC dough in doses ranging from 0.0 to 6.0 mg/kg, mice showed a dose-dependent reduction in consumption across access sessions, and this reduction was more rapid in adult mice at the highest doses, suggesting that adolescent mice might have been less sensitive to its aversive properties. These same mice, as well as a separate group of mice receiving injection (also 0.0 to 6.0 mg/kg THC), were given place conditioning sessions, alternating between THC dough and control dough or THC injection and vehicle injection, for 6 days per week and were tested once per week across a total of 3 weeks. Mice conditioned using edible THC showed a neutral response (neither reward nor aversion) at all doses. However, mice conditioned using injected THC showed a conditioned place aversion to the highest dose, which was more pronounced in adult mice. Interestingly, in mice self-administering edible THC, the dose of THC consumed was related to the outcome of place conditioning, such that a conditioned place preference was observed for adult mice which shifted their consumption of 3.0 mg/kg edible THC downward relative to those mice with full consumption of 3.0 mg/kg, and for adolescent mice which had the highest degree of consumption of 6.0 mg/kg edible THC relative to those mice with the lowest consumption of 6.0 mg/kg. Furthermore, initial place preference outcomes at the individual level at test 1 predicted subsequent doses of edible THC consumed, suggesting mice adjust their self-administration of edible THC based on the subjective experience it produces. Besides its impact in place conditioning, THC also had differential effects on body weight and locomotor activity based on age and route of administration. Collectively, this project demonstrates that adolescent mice are less sensitive to the hedonic properties of both cocaine and THC, and that differences in edible THC self-administration between ages, and between individuals within an age, are likely related the subjective experience of its rewarding and aversive properties.</div>

Hemp (Cannabis sativa L.) is a multi-use crop, able to provide fibre celulose a hurds for industrial treatment seeds for oil preparation biomass for energy conversion and produces secondary metabolites useful for pharmaceutical application. For its resistence to stress ability to accumulate high concentration of heavy metals and low cultivations demands, it can also be used for phytoextractions. Current research is focused on establishment of cultivation protocol, which allows transformation of callus cultures, and their regeneration with high efficiency. In this thesis, several varieties of hemp were transferred to in vitro conditions and were tested for their ability to form callus. The best results were achieved using the hypocotyl segments in a nutrient medium supplemented with 1 mg/L of naphtylacetic acid and one of these two synthetic cytokinins 0,5 mg/L of thidiazuron or 5 mg/L of 6-benzylaminopurine. No significant difference in the use of these two cytokinins were observed. None of the explants on four different test media for regeneration of shoots were able to succesfully regenerate. Transformation of hemp was tested using two different methods. Transformed protoplasts from hemp leafs after agroinfiltration were isolated. This method turn out to be unsuitable for use with hemp due to its...

College students have an increased risk for cannabis use, trauma exposure, and posttraumatic stress disorder (PTSD). Cannabis use disorder (CUD) and PTSD comorbidity is high, and given the negative consequences of the comorbidity (e.g., poor academic outcomes), there is a need to understand comorbid CUD-PTSD etiology. Two primary etiologic models exist: self-medication (i.e., PTSD à CUD) and high-risk (i.e., CUD à PTSD) hypotheses. This study 1) examined the prevalence and predictors of cannabis use and interpersonal trauma (IPT) exposure; 2) investigated the relationship between cannabis use and IPT; and 3) examined cannabis use, IPT, and PTSD through mediational self-medication and high-risk hypotheses lenses in a large (n = 9,889) longitudinal study of college students. Aim 1 found the prevalence of lifetime problematic (i.e., use ≥ 6 times) and experimental (i.e., use 1-5 times) cannabis use was 28.3% and 17.4%, respectively. Aim 1 results also estimated that the prevalence of lifetime IPT exposure was 35.9%. Aim 2 results supported the self-medication hypothesis, but not the high-risk hypothesis. Overall model fit from Aim 3 was poor. Nonetheless, Aim 3 results did not support the self-medication or high-risk hypotheses. Given the poor model fit of Aim 3, results should be interpreted with caution. However, as a whole, these findings provide preliminary support for the self-medication hypothesis, indicating that those reporting IPT exposure and probable PTSD may be at risk for cannabis use. Implications of these findings, in light of study limitations, are discussed.

No description available.

Interest in the use of medical cannabis for the treatment of drug-resistant epilepsy in children has grown over the last decade; however, little is known about its potential benefits and harms, cost-effectiveness, or the perspectives of key stakeholders. In this thesis, a health technology assessment approach was adopted to assess the intended and unintended consequences of medical cannabis use in the treatment of pediatric drug-resistant epilepsy. This thesis comprises three main sections: (1) a living systematic review of the benefits and harms of medical cannabis for the treatment of pediatric epilepsy, including drug-resistant forms; (2) an economic evaluation of the cost-effectiveness of medical cannabis for the treatment of pediatric drug-resistant epilepsy, and (3) qualitative exploration of the perspectives of neurologists and parents of children with drug-resistant epilepsy about the use of medical cannabis in this population. While neurologists generally perceived medical cannabis as a viable treatment option for drug-resistant epilepsy in children, particularly after other treatments have failed, they identified several gaps in the evidence base, including a lack of long-term studies and a lack of evidence related to cannabinoids other than cannabidiol. This is in keeping with the findings of the living systematic review, which support a beneficial role for medical cannabis in reducing seizures associated with drug-resistant epilepsy, although the certainty of the evidence was moderate at best. Parents described experiencing many barriers to accessing medical cannabis for their children, primarily related to finding a health care provider to authorize its use, the high cost of cannabis-based treatments, and a lack of reimbursement through public or private insurance programs. However, cannabinoid oil may be a more cost-effective treatment for some types of pediatric drug-resistant epilepsy compared with antiepileptic drugs currently reimbursed by some provincial insurance programs. These findings suggest that medical cannabis is a potentially effective and cost-effective treatment for drug-resistant epilepsy that may addresses an unmet need. However, additional studies are needed to address uncertainty related to the long-term benefits and harms of cannabis-based products, particularly with respect to products available in Canada.

This study investigated whether core beliefs about the world being safe and predictable (i.e. world assumptions) mediated the association between discrimination and internalizing and substance use problems among individuals from marginalized groups. Path analyses tested mediating effects of four types of world assumptions on the association between discrimination (race-, gender-, and sexual orientation-based) and anxiety, depression, alcohol and cannabis problems in college students (N = 1,181, agemean = 19.50, SD = 1.67). Limited support for mediation by world assumptions was found: among Asian students, race-based discrimination indirectly impacted anxiety symptoms through low perceived controllability of events. Direct effects across groups and discrimination types were also found.