Characterisation of cannabinoid receptors on immune cells and cell lines

Cannabinoids may inhibit immune cell function by modulating cytokine/chemokine release but the receptors mediating these events are poorly characterised. The aim of this thesis is to characterise cannabinoid receptors mediating cytokine/chemokine release from immune and inflammatory cells by measuring the effects of cannabinoids on cytokine release using ELISA technique. Apoptosis of inflammatory cells was also assessed by visual evaluation of cells treated with cannabinoids using a nuclear fluorochrome 4'6-diamidino-2 phenyl indole dihydrochloride (DAPI). Non-selective cannabinoid receptor agonists CP55,940 (10-6 -10-4 M- 10 'S M), A? - THC (10 -10 M) and anandamide (10 M- 10-4 M) inhibited LPS-induced release of TNF-a from THP-1 cells, a monocytic cell line. The cannabinoid CB2 receptor antagonist SR144528 (10 -6 M) but not the cannabinoid CB1 receptor antagonist SR141716A (10 -6 M) antagonised the inhibitory effects of CP55,940 (pA2 = 6.1 t 0.1, n=6) on THP-1 cells. Similarly, CP55,940 (10-6-104 M -10 'S M), 09-THC (10 10 M -10 -S M) and anandamide (10 -6 M -10'4 M) inhibited PHA/PMA-induced IL-2 release from Jurkat cells, a lymphocytic cell line. However in contrast to THP-1 cells, neither SR141716A (10 -6 M) nor SR144528 (10 -6 M) antagonised the inhibitory effects of CP55,940 on this cell line. In peripheral blood mononuclear cells a nonselective cannabinoid receptor agonist WIN55212-2 (10'10 M-10'5 M) and a selective cannabinoid CB2 receptor agonist JWH 015 (10 -10 M- 10 -S M) inhibited PHAinduced release of IL-2. These effects were antagonised by SR144528 (10-6 M) (pA2 = 6.3 ± 0.1; 6.5 ± 0.1, n=5 respectively) but not by SR141716A (10 -6 M). CP55,940 (10 -10 M -10 -5 M) produced a small, non-significant (P> 0.05) inhibitory effect on IL-2 release. 09-THC (10 -10 M-10-6 M) and ACEA (10 -'0 M- 10 -6 M) had no significant inhibitory effect on the release of IL-2 from PBMC. CP55,940 (10 M) and A9- THC (10 M) antagonised the inhibitory effects of WIN55212-2 (pA2 = 6.1 ± 0.1; 6.96 ± 0.16, n=5 respectively). In HT-29 cells, CP55,940 (10"10 -10"5 M- 10 M), A9-THC (10 -10 M -10 -5 M), WIN55212-2 (10"10 M-10-5 M) and JWH 015 (10 -10 M- 10 -5 M) inhibited IL-8 release. SR141716A (10 -6 M) antagonised the inhibitory effects of CP55,940 (pA2 = 8.3 ± 0.2 n=6) but did not antagonise the effects of WIN55212-2 and JWH 015. SR144528 (10 -6 M) but not SR141716A (10 -6 M) antagonised the inhibitory effects of CP55,940 (pA2 = 8.2 ± 0.8, n=6), WIN55212-2 (pA2 = 7.1± 0.3, n=6), JWH 015 (pA2 = 7.6 ± 0.4, n=6) respectively. A protein the size of cannabinoid CB2 receptors was localised in this cell line by Western blotting. CP55,940 and WIN55212-2 inhibited basal and agonist-evoked increases in both intracellular cyclic AMP and intracellular calcium at the same concentration as that inhibiting TNF-a-induced release of IL-8. Furthermore anandamide (>1 μM) but not CP55,940 caused apoptosis in Jurkat and HT-29 cell. These data suggest that activation of cannabinoid CB2 receptors in THP-1 cells, PBMC and HT-29 cells could lead to inhibition of cytokine/chemokine release. Furthermore,c annabinoid-evokedin hibition of basal and agonist stimulated increases in HT-29 cells may be related to cannabinoid-evokedin hibition of IL-8 release.T hus data presented in this thesis suggest that cannabinoid CB2 receptor agonists with high efficacy may have potential clinical utility in the treatment of inflammatory conditions such as inflammatory bowel disease (IBD) or chronic obstructive pulmonary disease (COPD) and other inflammatory disorders where epithelial cells have a major role

The impact of proliferation of private secondary schools on the quality of education in Ogun State

This study investigated the impact of proliferation of private secondary schools on the quality of education in Ogun State. To give direction to the study, two null hypotheses were postulated. The sample for the study comprised of 200 civil and public servants in the employment of Ogun State Government who were randomly selected. The instrument used for data collection was self- developed questionnaire. Data collected were analysed statistically with chi-square method at 95% confidence level. The results of data analyses showed that access to secondary education has not improved with proliferation of private secondary schools (X2 = 18.93; df = 12; P = 0.09 > 0.05) and that the proliferation of private secondary schools contributed to better performance of students in public certificate examinations (X2 = 24.56; df = 12; P = 0.00 < 0.05). Based on the findings of the study it was recommended among others that the private owners of secondary schools should see the establishment of their schools first of all as service to humanity by charging moderate fees for all and sundry to have access to quality education.Keywords: proliferation, private secondary schools, quality of educatio

ICT in National Pandemic: The Hope for Academic Development (Case Study of Secondary Schools and Universities in Owerri Municipal Nigeria)

Information and communication technologies (ICT) have become more of a classroom to all nation in carrying out academic activities. This research work used descriptive research method of analysis. This study was concerned with looking at the significance and hope of nations through the use of ICT in academic development in secondary and university education in Owerri municipal Imo state, Nigeria. Out of the Two hundred and five (205) respondent used for this research from different schools, including teachers the majority of respondents 132 (64.4%) were females and the remaining 73 (35.6%) were males. Their ages were 73 (36 %) for 16-20 years, 61 (30 %) for 10-15 years, 51 (26%) for 20-25 years and 17 (8 %) for 26 years above. All the 205 respondents indicated that ICT is the only hope for distance learning during national pandemic for academic development. The researcher went further to ask the respondents the frequently used ICT software that aids online learning and the highest respond shown that whatsapp (50 %) was the highest used app due to its ability to reduce data consumption followed by Google classroom (24%). They found out that the use of ICT encourage research activities since all information needed for any form of online research is found on the internet. Majority of the respondents strongly agreed that ICT permits online assignment (57.5 %). Communicating high expectations, emphasizing time on task, creating quizzes and exams, immediate feedback mechanisms are major principles underlying teaching. This research can conclude that ICTs are said to help expand approach to education, support the relevance of education to the progressively digital workplace, and raise educational quality. Keywords: academic, ICT, national, pandemic DOI: 10.7176/IKM/10-7-02 Publication date:October 31st 202

A Tale of Two Countries: An Assessment of the EU-US Trade Relationship in Agriculture

Although it is recognized that agriculture is not the most financially lucrative sector of EU and US economies, it has tremendous importance politically for these nations. But, agriculture is by far one of the most important features of developing countries economies. The US and the EU are two of the most prominent members of the World Trade Organization, and the WTO often relies on them to lead the charge towards open trade. But the inability of the EU and the US to agree on policy in agriculture has made the Doha Round negotiations in agriculture very difficult. This paper aims to show that the EU-US trade relationship in agriculture is equally economic as well as political, and that the fate of these rounds of WTO negotiations and future legislative reform in the US and the EU rely heavily on the EU and US ability to cooperate with each other

Ex Vivo Activity of Cardiac Glycosides in Acute Leukaemia

BACKGROUND: Despite years of interest in the anti-cancerous effects of cardiac glycosides (CGs), and numerous studies in vitro and in animals, it has not yet been possible to utilize this potential clinically. Reports have demonstrated promising in vitro effects on different targets as well as a possible therapeutic index/selectivity in vitro and in experimental animals. Recently, however, general inhibition of protein synthesis was suggested as the main mechanism of the anti-cancerous effects of CGs. In addition, evidence of species differences of a magnitude sufficient to explain the results of many studies called for reconsideration of earlier results. PRINCIPAL FINDINGS: In this report we identified primary B-precursor and T-ALL cells as being particularly susceptible to the cytotoxic effects of CGs. Digitoxin appeared most potent and IC(50) values for several patient samples were at concentrations that may be achieved in the clinic. Significant protein synthesis inhibition at concentrations corresponding to IC(50) was demonstrated in colorectal tumour cell lines moderately resistant to the cytotoxic effects of digoxin and digitoxin, but not in highly sensitive leukaemia cell lines. CONCLUSION: It is suggested that further investigation regarding CGs may be focused on diagnoses like T- and B-precursor ALL

Cannabinoid pharmacology in cancer research: A new hope for cancer patients?

Cannabinoids have been used for many centuries to ease pain and in the past decade, the endocannabinoid system has been implicated in a number of pathophysiological conditions, such as mood and anxiety disorders, movement disorders such as Parkinson's and Huntington's disease, neuropathic pain, multiple sclerosis, spinal cord injury, atherosclerosis, myocardial infarction, stroke, hypertension, glaucoma, obesity, and osteoporosis. Several studies have demonstrated that cannabinoids also have anti-cancer activity and as cannabinoids are usually well tolerated and do not produce the typical toxic effects of conventional chemotherapies, there is considerable merit in the development of cannabinoids as potential anticancer therapies. Whilst the presence of psychoactive effects of cannabinoids could prevent any progress in this field, recent studies have shown the value of the non-psychoactive components of cannabinoids in activating apoptotic pathways, inducing anti-proliferative and anti-angiogenic effects. The aforementioned effects are suggested to be through pathways such as ERK, Akt, mitogen-activated protein kinase (MAPK) pathways, phosphoinositide 3-kinase (PI3K) pathways and hypoxia inducible factor 1 (HIF1), all of which are important contributors to the hallmarks of cancer. Many important questions still remain unanswered or are poorly addressed thus necessitating further research at basic pre-clinical and clinical levels. In this review, we address these issues with a view to identifying the key challenges that future research needs to address

Physicochemical and microbiological analysis of canned and bottled fruit juices sold in Owerri Metropolis

This study was designed to investigate the physiochemical and microbiological analysis of canned and bottled fruit juices sold in Owerri metropolis. The samples were subjected to standard microbiological analysis and physiochemical parameters. The titratable acidity of the fruit juice samples ranged from 0.15% to 0.31%, while the ash content of all the samples varied from 0.32% to 0.63%. The total solid of all the samples was from 4.10% to 12.25%. The moisture content of the samples ranged from 2.32% to 4.81% and the pH was from 3.0 to 4.01. The total bacteria count (CFU/ml) of all the fruit juice samples showed to be from 1.1×102 to 4.1×102. Finally, the total fungi count (CFU/ml) varied from 0.0×102 to 1.2×102. Bacteria isolates from the samples includes Bacillus sp. (34.7%), followed by Enterococcus sp. (17.3%), Aspergillus sp. (21.7%) and Lactobacillus sp. (26.0%). Fungi isolates were identified as Penicillum sp. and Saccharomyces sp. The Standard Organization of Nigeria and NAFDAC should define more specifically the quality control standards of locally manufactured commercial juices, stating clearly the minimum and tolerable numbers and types of microorganisms

Microorganism Associated with Cassava Fermentation for Abacha Product Sold in Owerri, Imo State

The microorganism associated with cassava fermentation for producing Abacha was investigated using four different Farms in Owerri (Obinze, Umugwuma, Urata and Orji) as sample collection areas. Four of the cassava tubers that harvested, were washed, cooked and soaked with water after cutting to produce Abacha, which was then fermented using standard microbiological methods. The wastewater was subsequently sampled. All the samples A, B, C, and D showed significant bacteria growth after application of NA, MRS, PDA. In the study, six bacteria and fungi isolates were identified: Escherichia coli, Staphylococcus aureus, Lactobacillus spp, Candida spp., Aspergillus spp. and Bacillus spp. Results showed that the total heterotrophic bacteria count of the sample varies within 8.0×1010 to 2.5×1012 and total fungi count (TFC) ranges from 6.6×109 to 9.6×109. The study also showed that Lactobacillus spp. and Escherichia coli had the highest prevalence occurrence of 75.00% (P < 0.05), while Staphylococcus aureus, Saccharomyces spp., Bacillus spp. and Candida spp. had a percentage occurrence of 50.00%, respectively. There is, therefore, urgent need for public enlightenment on public health implications, need for proper hygiene, as well as need for strategies for preventing and controlling the microorganisms which do not play a role in the fermentation of cassava for Abacha production

Biochemical Manifestation of HIV Lipodystrophy Syndrome

Objectives Highly active anti-retroviral therapy (HAART), including protease inhibitors (PI) have led to dramatic improvements in the quality and quantity of life in patients with acquired immunodeficiency syndrome (AIDS). However, a significant number of AIDS patients on HAART develop characteristic changes in body fat redistribution referred to as lipodystrophy syndrome (LDS). Features of LDS include hypertrophy in the neck fat pad (buffalo hump), increased fat in the abdominal region (protease paunch), gynecomastia and loss of fat in the mid-face and extremities. Methods The aim of this paper is to review the current knowledge regarding this syndrome. This article reviews the published investigations on biochemical manifestation of HIV lipodystrophy syndrome. Results It is estimated that approximately 64% of patients treated with PI will experience this syndrome. Biochemically, these patients have increased triglycerides (Trig), total cholesterol (TC), low-density lipoprotein-cholesterol (LDL-C) and extremely low high-density lipoprotein-cholesterol (HDL-C). Conclusions and Public Health Implications It is hoped that awareness of this syndrome would aid in early diagnosis and better patient management, possibly leading to a lower incidence of cardiovascular complications among these patients. Key Words HIV Lipodystrophy Syndrome, Highly active anti-retroviral therapy, Nucleoside Reverse Transcriptase Inhibitors, buffalo hump

Production of Adhesive from Cassava Starch in Owerri, Imo State, Nigeria

In this work, the potentials of adhesives production from cassava starch were investigated. An adhesive product produced from Manihot utilissima was compared with a commercially available adhesive. Improvements in the properties of the cassava-based adhesive were achieved by investigating the effects of temperature and the addition of a viscosity enhancer/stabilizer (borax) on the density of the produced adhesive. The results obtained provide a wide range of insight into the production potentials of using cassava starch as a raw material in the production of large scale adhesives