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Artemisinins have low aqueous solubility that results in poor and erratic absorption upon oral administration. The poor solubility and erratic absorption usually translate to low bioavailability. Artemisinin-based monotherapy and combination therapies are essential for the management and treatment of uncomplicated as well as cerebral malaria. Artemisone and artemiside are novel artemisinin derivatives that have very good antimalarial activities. Pheroid™ technology is a patented drug delivery system which has the ability to entrap, transport and deliver pharmacologically active compounds. Pharmacokinetic models were constructed for artemisone and artemiside in Pheroid™ vesicle formulations. The compounds were administered at a dose of 50.0 mg/kg bodyweight to C57 BL/6 mice via an oral gavage tube and blood samples were collected by means of tail-bleeding. Drug concentrations in the samples were determined using an LC/MS/MS method. There was 4.57 times more artemisone in the blood when the drug was entrapped in Pheroid™ vesicles in comparison to the drug only formulation (p < 0.0001). The absorption of artemiside was not dramatically enhanced by the Pheroid™ delivery system. http://dx.doi.org/10.1016/j.ijpharm.2011.05.003

BackgroundThe molecular basis of Parkinson's disease in South African population groups remains elusive. To date, substitutions in the GBA gene are the most common large-effect genetic risk factor for Parkinson's disease. The primary objective of this study was to determine the prevalence of GBA substitutions in South Africans with idiopathic Parkinson's disease.MethodsParticipants were recruited from tertiary hospitals in the Gauteng Province in South Africa. All participants were screened for substitutions in GBA exon 8-11 and the full coding region was analysed in 20 participants. Peripheral -glucocerebrosidase enzymatic activity of GBA-carriers was measured in mixed leukocytes.ResultsOf 105 Caucasian Parkinson's disease participants (82.7% Afrikaner) with an average age of disease onset of 61.912.2years and 40 controls (age 73.4 +/- 12.4years) were included. Heterozygous GBA substitutions were identified in 12.38% of affected participants (p.G35A, p.E326K, p.I368T, p.T369M, p.N370S, p.P387L and p.K441N) and 5.00% of controls (p.E326K and p.T369M). The substitutions ranged from predicted benign to moderately damaging; with p.E326K and p.T369M most prevalent, followed by the Afrikaner Gaucher disease substitution p.P387L. Severe Gaucher disease mutations, like p.L444P, were absent in this cohort. Enzyme activity analysis revealed a nonsignificant reduction in the GBA-Parkinson's disease individuals (14.49 +/- 2.30nmol/h/mg protein vs. 15.98 +/- 3.06nmol/h/mg in control samples). GBA substitutions occur in both young-onset and late-onset Parkinson's cases in the cohort.ConclusionMild GBA substitutions that may not cause Gaucher disease were a common risk factor for Parkinson's disease in the participant group.

The purpose of this reported study was to develop and validate an LC-MS/MS method for the quantification of goserelin in a Pheroid (R) formulation simulated intestinal fluid. Biopharmaceuticals are formulated in drug delivery systems to improve their gastrointestinal stability. Goserelin, a peptide drug was formulated in Pheroid (R) delivery system and its gastrointestinal stability assessed using simulated intestinal fluid, which required an assay to determine the varying amounts of goserelin remaining after a specific time. Several extraction methods and solvents investigated to extract goserelin from complex matrix led to either poor recovery, peak shape or high background interference. A rapid gradient reversed-phase method coupled to tandem mass spectrometry detection was optimized for the separation and quantification of the extracted peptide. A simple, reproducible and good recovery extraction procedure for goserelin quantification was achieved through simultaneous acetonitrile protein precipitation and water-saturated n-butanol liquid-liquid extraction with water dilution. The method was found to be rapid, specific, precise and accurate, and successfully applied to determine goserelin remaining content in a simulated intestinal fluid, with potential use in other lipid-based formulation evaluated in simulated intestinal fluids. (C) 2019 Elsevier B.V. All rights reserved

GBA mutations are to date the most common genetic risk factor for Parkinson's disease. The GBA gene encodes the lysomal hydrolase glucocerebrosidase. Whilst bi-allelic GBA mutations cause Gaucher disease, both mono- and bi-allelic mutations confer risk for Parkinson's disease. Clinically, Parkinson's disease patients with GBA mutations resemble idiopathic Parkinson's disease patients. However, these patients have a modest reduction in age-of-onset of disease and a greater incidence of cognitive decline. In some cases, GBA mutations are also responsible for familial Parkinson's disease. The accumulation of alpha-synuclein into Lewy bodies is the central neuropathological hallmark of Parkinson's disease. Pathologic GBA mutations reduce enzymatic function. A reduction in glucocerebrosidase function increases alpha-synuclein levels and propagation, which in turn inhibits glucocerebrosidase in a feed-forward cascade. This cascade is central to the neuropathology of GBA-associated Parkinson's disease. The lysosomal integral membrane protein type-2 is necessary for normal glucocerebrosidase function. Glucocerebrosidase dysfunction also increases in the accumulation of O-amyloid and amyloid-precursor protein, oxidative stress, neuronal susceptibility to metal ions, microglial and immune activation. These factors contribute to neuronal death. The Mendelian Parkinson's disease genes, Parkin and ATP13A2, intersect with glucocerebrosidase. These factors sketch a complex circuit of GBA-associated neuropathology. To clinically interfere with this circuit, central glucocerebrosidase function must be improved. Strategies based on reducing breakdown of mutant glucocerebrosidase and increasing the fraction that reaches the lysosome has shown promise. Breakdown can be reduced by interfering with the ability of heat-shock proteins to recognize mutant glucocerebrosidase. This underlies the therapeutic efficacy of certain pharmacological chaperones and histone deacetylase inhibitors. These therapies are promising for Parkinson's disease, regardless of mutation status. Recently, there has been a boom in studies investigating the role of glucocerebrosidase in the pathology of Parkinson's disease. This merits a comprehensive review of the current cell biological processes and pathological pictures involving Parkinson's disease associated with GBA mutations.

Safety and efficacy are of concern in developing new compounds. There is a paucity on the changes made by new compounds on inflammatory cytokines in combination with oxidative stress markers. Cytokines are potential markers of drug safety, focusing on cardiovascular, neurologic, and respiratory functions. In addition oxidative stress markers may indicate the impact of drugs on physiological processes. According to literature, a dysregulation of cytokines can be indicative of chronic inflammation. There is no formal reference of normal ranges of inflammatory markers. By the establishment of a baseline for preclinical safety studies, the effects of new compounds on cytokines can be quantified. When cancer cells were then exposed to an anti-cancer compound, the anti-oxidative enzymes' activity increased. Oxidative stress plays an important role in the regulation of physiological processes. The aim of this study was to determine the changes in the cytokines and anti-oxidant enzymes in the renal and hepatic systems in vivo (xenograft rats) after treatment with radiolabeled compounds (confidential, under development). Tissue was collected from xenografted RNU rats, from a control group and two radiolabeled compound exposed groups. The cytokines were analyzed using a CBA Flow Cytometry cytokine assay. Oxidative stress markers analyzed was intracellular reactive oxygen species (ROS), superoxide dismutase (SOD), catalase (CAT), protein carbonyls (PC), and lipid peroxidation (malondialdehyde, (MDA)). There were changes in the cytokines and oxidative stress markers post-exposure to radiolabeled compounds. In the kidneys, the copper-chelate (Cu-c) increased the pro- and anti-inflammatory cytokines (55% & 35%), where Pd-c decreased both (30% & 39%). In contrast liver cytokines for the Cu-c group (19 & 41%) decreased but increased for Pd-c (45 & 84%). In the kidneys, ROS was lower in Cu-c groups and showed an increase for Pd-c. The SOD levels in the kidneys was increased (30%) for Cu-c and decreased (20%) for Pd-c. A 120% increase for Cu-c and 328% decrease for Pd-c was seen for SOD in the liver. The CAT levels in the kidneys were elevated for Cu-c (45%) and Pd-c (67%) compared to the control. In contrast CAT levels in the liver showed a 50% decrease in both Cu-c and Pd-c. The Cu-c group was responsible for more protein damage, whereas Pd-c was responsible for more lipid peroxidation. A relationship between the anti-oxidant markers and the cytokines was seen. This suggest that cytokines in combination with oxidative stress markers may be used as indicators for drug safety

Lung cancer is the leading cause of cancer related deaths, globally, with poor survival profile. Cisplatin is frequently used in the treatment of many solid tumors including lung cancer despite its toxicity; ototoxicity, peripheral neuropathy, and nephrotoxicity. Turmeric and ginger extracts constitute a mix of biologically active compounds such as curcumin and [6]-shogaol with established chemopreventive, anti-inflammatory and antioxidant properties. Human clinical safety data show that turmeric and ginger extracts exhibit low intrinsic toxicity even at higher doses. The present in vitro safety study investigated the chemopreventive activity of combined Meriva® (a curcumin phytosome) and ginger extract in Pheroid® delivery system against the A549 lung cancer cell line. Pheroid® is a colloidal emulsion system comprised of plant and ethyl esters of essential fatty acids (EFAs) as the dispersed phase and nitrous oxide saturated water as the continuous phase. Six combinations of Meriva® and ginger extract were formulated in Pheroid®. The contents of curcumin in Meriva® and [6]-shogaol in ginger extract were quantified using LC-MS/MS. Cell viability was studied using MTT and neutral red uptake assays. Cellular uptake as well as safety in terms of induction of oxidative stress markers including reactive oxygen species (ROS), superoxide dismutase (SOD) and catalase (CAT) were examined between Pheroid® formulated phytochemical combination and cisplatin. Pheroid® enhanced cellular uptake of Meriva® and ginger extract combination and proved to be safe despite higher concentration of the actives were present in the cells. Meriva® and ginger extract at 1:1.43 combination ratios in Pheroid® had a comparable anticancer activity with cisplatin. In the oxidative stress test, cisplatin induced significantly higher ROS and lower CAT levels, and insignificantly lowered SOD level compared to the control while the same Meriva® and ginger extract combination in Pheroid® showed insignificantly lower ROS level and significantly lower SOD and CAT levels compared to the control. The content of curcumin in Meriva® and [6]-shogaol in ginger extract were found to be 400 mg/g and 10.61 mg/g, respectively. Although cisplatin is an effective antineoplastic agent, its side effects stem from its nonselective induction of oxidative stress to both cancer and normal cells. Pheroid® formulated Meriva® and ginger extract combination demonstrated a comparable anticancer activity with cisplatin without induction of oxidative stress and can therefore be used as a safe and effective adjuvant therapy in lung cancer

Graphical abstract, Highlights • Acute administration of 2000 mg/kg of the Pheroid® delivery system was tolerated upon intravenous administration in BALB/c mice and Sprague-Dawley rats. • Oral administration was tolerated in both acute toxicity evaluation (14-days post single dose administration) and during chronic administration (90-days dosing). • No mutagenicity was present during the Ames assay. • A statistically significant increase in creatinine levels in the sub-chronic female treatment group was observed, however no treatment related pathology was identified during histopathology. • This evaluation did not identify any risk factors present for toxicity during oral or intravenous administration of the tested formulations during acute or repeated dosing., The Pheroid® drug delivery system is now on the threshold of progressing into human clinical trials for various patented pharmaceutical applications and a systematic investigation of its toxicological properties in vitro and in vivo is thus a priority. Colloidal dispersions (nano- and microemulsions) demonstrate the ability to be adapted to accommodate either lipophilic, hydrophilic or amphiphilic drug molecules. The colloidal dispersions investigated during this evaluation has a general size of 200 nm - 2 μm, a zeta-potential of -25 mV and the main ingredient was ethyl esters of essential fatty acids. The Ames mutagenicity assay was performed on selected Salmonella thyphimurium strains TA98, TA100 and TA102. The Ames assay included S9 metabolic activation and no mutagenicity was present during the assay. The effect of acute and subchronic administration on a biological system was investigated in two species of rodent (BALB/c mice and Sprague-Dawley rats). Observations focused on the physical condition, blood biochemical analysis and the haematological profiles. Gross necropsy was performed on all the test animals. Organ weights followed by histopathology of selected organ tissues were recorded. During the acute evaluation animals showed tolerance of the maximum prescribed dose of 2000 mg/kg (according to OECD guidelines) in two rodent species after intravenous administration (absolute bioavaibility). The oral formulation was tolerated without incidents in both acute and subchronic studies. Although valuable baseline safety data was obtained regarding the Pheroid® system, future studies with the entrapped active pharmaceutical ingredients is necessary to provide a definitive safety profile.

Southern Africa (South Africa as well as neighboring countries) has not had an infrastructure for testing the safety of new drugs or other therapeutic entities, including plant-based or herbal treatments coming from the indigenous cultures, according to the current practice and regulatory requirements. Simultaneously, Southern Africa is challenged by very high rates of immune deficiency diseases which have further led to the resurgence of diseases such as tuberculosis, which is commonly thought to be eradicated, at least in Western cultures. Thus, there is high medical need for affordable, effective and safe treatments, but up to now there has been no local expertise and capabilities for state-of-the-art drug testing. Based on an initiative funded by the South African Department of Science and Technology, the Potchefstroom campus of North-West University was chosen for the establishment of a center of excellence for doing rodent-based drug testing research. The centerpiece of the Pre-Clinical Drug Development Platform (PCDDP) is a new rodent vivarium for breeding of SPF rats and mice and associated laboratories for conducting a wide range of physiological and pharmacological studies. Notably, the vivarium has become the first AAALAC-accredited animal facility in Sub-Sahara Africa and is qualified for conducting GLP studies. The establishment of safety pharmacology models is a first priority of the PCDDP, including expertise in studies to examine electrophysiological effects in vitro, and potential CNS, cardiovascular and respiratory effects. The intention is to offer the rodent-based safety pharmacology testing not only to local academic and company-related scientists, but also to those throughout South Africa and neighboring countries.

Tuberculosis, which typically presents as a pulmonary disease, has a complex pathology. The primary site of infection, the Ghon focus, recruits immune cells and a granuloma forms. At earlier stages the granuloma is still vascularized, offering the best opportunity for drug treatment. In the more progressive state blood flow is reduced and a distinct caseous structure develops. Effective delivery of drugs to bacilli in the core of the granuloma becomes very difficult. It is perceivable that granuloma cores could create conditions where bacilli persist and develop resistance. In this study we analyze drug delivery to granulomas by means of a nanoparticle delivery system. The model consists of two parts; the overall distribution of the nanoparticles is described by a simple circulatory model and this result is used in the second part, focusing on transport in a capillary lined with macrophages. Nanoparticles enter the macrophages where they are metabolized and the drugs are released. The model reveals significant differences in drug concentrations between the plasma and macrophages. Based on the results of the model, strategies for improved drug delivery are proposed.

Global tuberculosis (TB) control and eradication is hampered by the unavailability of simple, rapid and affordable diagnostic tests deployable at low infrastructure microscopy centers. We have developed and evaluated the performance of a nucleic acid amplification test for detection of Mycobacterium tuberculosis (MTB), the NWU-TB test, in clinical sputum specimens from 306 patients with suspected pulmonary tuberculosis. The test involves sputum sample processing using a Lyser device within 7 min, followed by rapid multiplex-PCR on a fast thermal cycler within 25 min, and amplicon resolution on agarose gel electrophoresis. Samples were also examined for presence of MTB using smear microscopy, GeneXpert and MGIT culture. Results were assessed in comparison to a MGIT culture as gold standard. Of the 306 patients, 174 had a previous TB history or already on treatment, and 132 were TB naive cases. The NWU-TB system was found to have an overall sensitivity and specificity of 80.8% (95% CI: 75–85.7) and 75.6% (95% CI: 64.9–84.4) respectively, in comparison to 85.3% (95% CI: 79.9–89.6) and 73.2% (95% CI: 62.2–82.4) respectively for GeneXpert; and 62.1% (95% CI: 55.3–68.4) and 56.1% (95% CI: 44.7–67) respectively for smear microscopy. The study has shown that the NWU-TB system allows detection of TB in less than two hours and can be utilized at low infrastructure sites to provide quick and accurate diagnosis at a very low cost.

Huntington’s disease (HD) is a fatal genetic disease which causes uncontrolled movements by affecting the striatum of the brain. Genetics only partially predict the age of onset. Firstly, the research showed that in a cohort of 13.856 HD-carriers, persons with birth complications were diagnosed up to 6 years earlier. The most common birth complication among these persons: perinatal asphyxia (PA), increased nitrosidative stress in the striatum of a rat model. Nitrosidative stress is also an important factor in HD and could contribute to the gene-environment interaction found. Lastly, we showed that treatment with bone-marrow derived stem cells shortly after PA reduced the degree of disability in adulthood.

Colloidal suspensions of 14 nm gold nanoparticles (AuNPs) were repeatedly administered intravenously at three dose levels (0.9, 9 and 90 µg) to male Sprague Dawley rats weekly for 7 weeks, followed by a 14-day washout period. After sacrificing, the amount of gold was quantified in the liver, lungs, spleen, skeleton and carcass using neutron activation analysis (NAA). During the study, pre- and post (24 h) administration blood samples were collected from both the test and control groups, the latter which received an equal injection volume of normal saline. General health indicators were monitored together with markers of kidney and liver damage for acute and subchronic toxicity assessment. Histopathological assessments were done on the heart, kidneys, liver, lungs and spleen to assess any morphological changes as a result of the exposure to AuNPs. The mass measurements of all the groups showed a steady increase with no signs of overt toxicity. The liver had the highest amount of gold (µg) per gram of tissue after 56 days followed by the spleen, lungs, skeleton and carcass. Markers of kidney and liver damage showed similar trends between the pre and post samples within each group and across groups. The histopathological examination also showed no hepatotoxicity and nephrotoxicity. There was accumulation of Au in tissues after repeated dosing, albeit with no observable overt toxicity, kidney or liver damage.

Malaria and HIV are among the most important global health problems of our time and together are responsible for approximately 3 million deaths annually. These two diseases overlap in many regions of the world including sub-Saharan Africa, Southeast Asia and South America, leading to a higher risk of co-infection. In this study, we generated and characterized hybrid molecules to target Plasmodium falciparum and HIV simultaneously for a potential HIV/malaria combination therapy. Hybrid molecules were synthesized by the covalent fusion of azidothymidine (AZT) with dihydroartemisinin (DHA), a tetraoxane or a 4-aminoquinoline derivative; and the small library was tested for antiviral and antimalarial activity. Our data suggests that compound 7 is the most potent molecule in vitro, with antiplasmodial activity comparable to that of DHA (IC50 = 26 nM, SI >3000), a moderate activity against HIV (IC50 = 2.9 μM; SI >35) and not toxic to HeLa cells at concentrations used in the assay (CC50 >100 μM). Pharmacokinetics studies further revealed that compound 7 is metabolically unstable and is cleaved via O-dealkylation. These studies account for the lack of in vivo efficacy of compound 7 against the CQ-sensitive Plasmodium berghei N strain in mice, when administered orally at 20 mg/kg. http://dx.doi.org/ 10.1016/j.bmc.2012.06.038

Radiopharmaceuticals containing the motive tripeptide arginyl-glycyl-asparatic acid (RGD) are known to target ανβ3 integrins during tumor angiogenesis. A more generic kit radiolabeling procedure accommodating Ga-68 from different generators was developed for NOTA-RGD and evaluated for its versatile use and safety in subsequent in vivo applications. The [68Ga]NOTA-RGD kit was further verified for its expected biodistribution and pharmacokinetics in nonhuman primates and its clinical sensitivity to detect solitary pulmonary nodules (SPN) in cancer patients. Single vial kits containing 28–56 nmol of NOTA-cyclo-Arg-Gly-Asp-d-Tyr-Lys (NOTA-RGD) and sodium acetate trihydrate buffer were formulated. Versatility of the NOTA-RGD radiolabeling performance and adaption to a TiO2- and a SnO2-based generator type, characterization and long-term storage stability of the kits were carried out. The blood clearance and urine recovery kinetics as well as the image-guided biodistribution of [68Ga]NOTA-RGD was studied in a vervet monkey model. [68Ga]NOTA-RGD kits were further tested clinically to target solitary pulmonary nodules. The kits could be successfully formulated warranting integrity over 3–4 months with a good [68Ga]NOTA-RGD radiolabeling performance (radiochemical purity >95 %, decay corrected yield 76–94 %, specific activity of 8.8–37.9 GBq/μmol) The kits met all quality requirements to be further tested in vivo. [68Ga]NOTA-RGD cleared rapidly from blood and was majorly excreted via the renal route. The liver, spleen, heart and intestines showed initial uptake with steadily declining tissue activity concentration over time. In addition, the [68Ga]NOTA-RGD kit allowed for delineation of SPN from non-malignant lung tissue in humans. A more versatile radiolabeling procedure using kit-formulated NOTA-RGD and different generator types was achieved. The uncompromised in vivo behavior and efficient targeting of SPN warrants further investigations on the clinical relevance of [68Ga]NOTA-RGD derivatives to implement initial guidelines and management of patients, with regard to integrin targeted imaging.

Background: Mycobacterium tuberculosis (M. TB) remains the prime cause of bacterial mortality and morbidity world-wide. Therefore, effective delivery and targeting of drug to the cellular tropics is essentially required to generate significant results for tuberculosis treatment. The aim of the present study was to develop and characterize ligand anchored pH sensitive liposomes (TPSL) as dry powder inhaler for the targeted delivery of drugs in the target site i.e. lungs. Method: Ligand anchored PSL (TPSL) was prepared by thin film hydration for the combined delivery of Isoniazid (INH) and Ciprofloxacin HCl (CIP HCl) using 4-aminophenyl-α-D mannopyranoside (Man) as surface functionalized ligand and characterized using different parameters. Results: It was observed that size of the ligand anchored liposomes (TPSL) was slightly more than the non-ligand anchored liposomes (PSL). Drug release was studied at different pH for 24 hrs and it was observed that liposomes exhibited slow release at alkaline pH (58-64%) as compared to macrophage pH (81-87%) where it increased dramatically due to the destabilization of pH sensitive liposome (PSL). In vitro cellular uptake study showed that much higher concentration was achieved in the alveolar macrophage using ligand anchored liposomes as compared to its counterpart. In vivo study showed that maximum drug accumulation was achieved in the lung by delivering drug using ligand anchored PSL as compared to conventional PSL. Conclusion: It was concluded that ligand anchored pH sensitive liposome is one of the promising systems for the targeted drug therapy in pulmonary tuberculosis.

National malaria treatment policies are devised to guide health professionals and to facilitate diagnosis and case management. Following the recommendations of the WHO, Cameroon changed its malaria treatment policy from monotherapy to artemisinin-based combination therapy (ACT) as the first-line treatment for uncomplicated malaria. We report an investigation into trends of case management following this change in policy. Data was collected retrospectively, through consultation and perusal of laboratory and prescription registers of the Mbakong Health Centre. Analysis of data was done using SPSS and SAS Statistics. Data presented herein demonstrate that from 2006 to 2012, a total of 2484 (58.7%) of the total prescriptions included an anti-malarial, 1989 (47.0%) included an antibiotic and 1935(45.7%) included an antipyretic. The anti-malarials prescribed were Anti-malaria combination therapy (ACT) – 1216 (47.6%), quinine 1044 (40.8%) or SP 296 (11.6%). Of the 1216 patients prescribed an ACT, 441(36.3%) had a positive malaria parasite confirmation, 746 (61.3%) were negative for plasmodium. Overall, 29 patients (2.4%) were treated either with an ACT without any test performed. Quinine intake was recorded in 566 (54.2%) patients positive for plasmodium. ACT prescription increased from 23% in 2007 to between 44 and 45% in 2008–2009. During this period there was a corresponding drop in the prescription of quinine from 38% in 2007 to 13% in 2009 (r = −0.43, p > 0.05). Sulphadoxine-Pyrimethamine (SP) was restrictively prescribed to women of childbearing age (97.0%) after 2008. Antibiotics prescription dropped from 53.7% to 39.3% from 2010 to 2012. The odds of being prescribed an antibiotic was significantly higher in patients with a malaria negative result compared to malaria positive patients (OR = 6.12, CI 4.74–7.91, p < 0.00001). Overall, there is an over treatment of malaria, thus departing from the WHO guidelines of appropriate treatment. Although there is an overall increase in the prescription of ACT, less prescription of quinine and a noticeable restrain from prescription of SP to febrile cases, the old practice was still rampant. There is need for healthcare workers to adhere to guidelines in order to enhance the rational use of drugs to achieve appropriate treatment of uncomplicated malaria according to WHO guidelines DST/NWU Preclinical Drug Development Platform of the North-West University, the Division of Research,Catholic University of Cameroon Bamenda, the Research Office of the North-West University, Potchefstroom, South Africa, the Laboratory of Public Health Research Biotechnologies and the Njongdong Foundation

Pheroid® technology was assessed as an alternative to Freund's adjuvant to raise antibodies in experimental animals. Chickens were immunized with two recombinantly expressed Plasmodium falciparum proteins, lactate dehydrogenase (PfLDH) and glyceraldehyde-3-phosphate dehydrogenase (PfGAPDH), alone or in combination with Freund's adjuvant or Pheroid®. Chicken egg yolk antibodies (IgY) were isolated and compared for specificity, sensitivity and yield. Freund's adjuvant and Pheroid® stimulated prolonged antibody responses in chickens against both antigens. Affinity purified antibodies had specificity for the recombinant and the native proteins on Western blots. Antibodies generated in the presence of Freund's adjuvant had high sensitivity for both antigens. Pheroid® generated antibodies that detected the lowest concentration of recombinant PfLDH. Freund's adjuvant and Pheroid® both improved chicken IgY yields, with Pheroid® showing a 2-fold increase relative to controls. Pheroid® was well-tolerated in chickens and has potential for development as a safe adjuvant for testing alternative stimulatory factors to improve adjuvant formulations.

Objectives: The objectives were to determine the pharmacokinetics (PK) of artemisone and artemisone formulated in the Pheroid® drug delivery system in primates and to establish whether the formulation affects the in vitro metabolism of artemisone in human and monkey liver and intestinal microsomes. Methods: For the PK study, a single oral dose of artemisone was administered to vervet monkeys using a crossover design. Plasma samples were analyzed by means of liquid chromatography-tandem mass spectrometry. For the in vitro metabolism study, clearance was determined using microsomes and recombinant CYP3A4 enzymes, and samples were analyzed by means of ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry. Results: Artemisone and M1 plasma levels were unexpectedly low compared to those previously recorded in rodents and humans. The in vitro intrinsic clearance (CLint) of the reference formulation with monkey liver microsomes was much higher (1359.33 ± 103.24 vs 178.86 ± 23.42) than that of human liver microsomes. The in vitro data suggest that microsomal metabolism of artemisone is inhibited by the Pheroid delivery system. Conclusions: The in vivo results obtained in this study indicate that the Pheroid delivery system improves the PK profile of artemisone. The in vitro results indicate that microsomal metabolism of artemisone is inhibited by the Pheroid delivery system http://dx.doi.org/10.1517/17425255.2014.885503 http://www.tandfonline.com/loi/iemt20#.VjiobytpETY South African Technology Innovation Agency (TIA), the Swiss South African Joint Research Project Initiative and the North-West University

The in vitro antimalarial activities of artemisone and artemisone entrapped in Pheroid vesicles were compared, as was their ability to induce dormancy in Plasmodium falciparum . There was no increase in the activity of artemisone entrapped in Pheroid vesicles against multidrug-resistant P. falciparum lines. Artemisone induced the formation of dormant ring stages similar to dihydroartemisinin. Thus, the Pheroid delivery system neither improved the activity of artemisone nor prevented the induction of dormant rings.