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The International Mouse Phenotyping Consortium reports the generation of new mouse mutant strains for over 5,000 genes from targeted embryonic stem cells on the C57BL/6N genetic background. This includes 2,850 null alleles for which no equivalent mutant mouse line exists, 2,987 novel conditional-ready alleles, and 4,433 novel reporter alleles. This nearly triples the number of genes with reporter alleles and almost doubles the number of conditional alleles available to the scientific community. When combined with more than 30 years of community effort, the total mutant allele mouse resource covers more than half of the genome. The extensively validated collection is archived and distributed through public repositories, facilitating availability to the worldwide biomedical research community, and expanding our understanding of gene function and human disease.

Resistance against the current TB treatment regimens is spreading. A combination of the current TB regime with a novel drug delivery system, might lead to reduced resistance, shorter treatment durations and increased patient compliance. A delivery system is used to facilitate the delivery of specific therapeutically active ingredients to a targeted area in an organism (Vonarbourg et al., 2006). An ideal delivery system ought to be biodegradable, stable, pharmaceutically acceptable and suitable for targeted delivery (Buszello & Müller, 2000). The Pheroid® drug delivery system is such a system. It is an intricate polydisperse technology that is based on different colloidal emulsion systems (Grobler, 2009). In this study a novel Pheroid® drug delivery system was used with the addition of the mycobactericidal eicosapentaenoic fatty acid (EPA) and a targeting ligand Mycolic acid (MA). The in vivo anti-TB activity of the novel formulation alone, and in combination with the current treatment regime was determined against the H37Rv Mtb strain in the Kramnik mouse model. After inoculation, three groups of mice (n = 56) each received a different treatment formulation. Group 1 received the first line treatment API's (positive control), group 2 novel Pheroid® formulation and group 3 novel Pheroid® formulation in combination with the first line treatment API's. One group, group 4 (negative control), was infected but received no treatment. Six animals per treatment group were euthanised at different time intervals, i.e. at week 1, 2, 3, 4, 8 and lastly at week 12 and four animals in group 4 at different time intervals, i.e. at week 1, 8 and 12. Lungs were harvested for bacteriological examination in order to provide information regarding disease progression. Inflammation in the lungs indicates the presence of the Mtb infection, a direct correlation can be made between the mass of the lungs and the severity of the infection. Preliminary results obtained from the average mass of the lungs, showed that both group 1 and 3 suppressed the Mtb infection to a similar degree. Mice in these groups recovered quicker than those in groups 2 and 4. Some of the mice in group 2 and 4 reached the humane endpoint before the predetermined euthanasia dates, indicating an aggressive progression of the disease in these two groups. The preliminary results showed that the Mtb infection was suppressed in both treatment groups, group 1 and 3, while the carrier control (group 2) and the negative control (group 4) had no effect on the disease progression. Next the colony-forming units in the lung samples will be determined to give a more accurate quantitative measurement of the bacteriological infection. Further research will determine the relapse rate in the different study groups

Cannabidiol (CBD), a phytocannabinoid isolated from the Cannabis sativa plant, has been investigated for its medicinal properties including anti-psychotic, anti- emetic, anti-inflammatory, anxiolytic and anti- convulsant effects1,2. However, the per oral CBD bioavailability has been shown to be only 6%; presumably due to the high lipophilic nature and the extensive first pass metabolism of cannabinoids2,3. The lipid- based Pheroid® drug delivery system is a nano- and micro-sized particle delivery system that has previously been used to increase the bioavailability of pharmaceutical compounds such as certain hormonal, antimicrobial, antimalarial and antituberculosis drugs4,5,6,7.8. Recently, we have demonstrated in an in vivo preclinical study the ability of the Pheroid® to increase the oral bioavailability of CBD in rats compared to existing CBD products. A phase 1 clinical trial to evaluate the oral bioavailability of a Pheroid®-CBD formulation in humans is now underway. (See Table 1.) Preclinical study The bioavailability of four oral CBD formulations and one novel pro-Pheroid® -CBD formulation was compared in nine weeks old healthy Sprague Dawley rats (250 ± 53 g) where n = 8 rats per formulation. Each rat received 20 mg/kg of the relevant CBD product through oral gavage followed by consecutive blood sample collections (200 μL each) for 24 h. The CBD plasma concentration was determined for each sample by LC-MS/MS analysis and used to calculate the pharmacokinetic parameters of each formulation, i.e. Ke, Cmax, Tmax, T1/2 and AUC0–24. Clinical study The bioavailability of a test formulation (CBD-pro- Pheroid®) and a reference compound (CBD alone) will be compared in a singledose, double-blind phase I clinical trial with 12 healthy participants, using a randomized cross-over study design with a three-week wash-out period. Consecutive blood samples will be collected from each participant for 96 h per study arm. LC-MS/MS analysis will be used to determine the CBD plasma concentration in order to calculate the pharmacokinetic parameters, i.e. Ke, Cmax, Tmax, T1/2 and AUC0–96. Results of preclinical study The CBD-pro-Pheroid® formulation yielded a higher CBD C max (157.04 ± 66.58 ng/mL), AUC0–24 (1093.32 ± 470.32 ng/mL.hrs) than most of its comparators without the Pheroid® drug delivery system. Promising in vivo preclinical data showed an increased CBD bioavailability when comparing a novel pro- Pheroid®-CBD formulation to existing CBD formulations and formulations under development. The pharmacokinetic parameters of a similar pro-Pheroid®- CBD formulation in humans will be determined in the current study

In developing new compounds, questions about safety and efficacy are raised. Conventional preclinical studies of new compounds focus on bio-distribution, toxicology, safety pharmacology, pharmacokinetics, efficacy etc., but little information regarding the effect of these compounds on inflammatory cytokines are available. Cytokines can be used as markers to identify drug safety, focusing on cardiovascular, neurologic, and respiratory functions. Cytokines are produced by different cell groups, mainly by macrophages and helper T cells3 . Cytokines can be divided into two main groups: antiinflammatory cytokines and pro-inflammatory cytokines. Cytokines function as a complex network to control each other's responses and production4 . According to literature, a dysregulation of inflammatory markers or biomarkers can be indicative of chronic inflammation 1,2. There is no data available on the inflammatory cytokine values in healthy people and there is no formal reference of normal ranges of inflammatory markers. By establishing a baseline, it can be used in preclinical studies to determine the effects of new compounds on cytokines. The aim of this study is to determine baseline inflammatory cytokines concentration in an established in vivo model to eventually determine the effects of new radiolabeled compounds on the cytokines. Using an established xenograft model, a control baseline, a disease control (cancer), and drug treatment (radiopharmaceutical) cytokine levels will be determined by cytokine analyses of collected blood serum and tissues Antigenix America Inc.'s SUPER –X PLEX™ Flow Cytometry cytokine assays will be used on the BD Accuri® C6 Flow Cytometer. This multiplex assay has multiple bead populations, which are differentiated by the levels of fluorescence intensity and size. This allows for a distinct bead populations on the flow cytometer data output. It also makes it possible to measure multiple analytes with a single reaction. The following results are anticipated: Determining the individual baseline concentrations of the inflammatory cytokines. To observe a clear change in cytokine profiles between the baseline, disease control and drug treatmen

One of the most active, widely used and well-known drugs in breast cancer chemotherapy, doxorubicin (DOX), is an anthracycline ring antibiotic derived from the bacteria Streptomyces peucetius. Its clinical use is however, hampered by dangerous side effects like cumulative dose-dependent cardiotoxicity (leading to myocardiopathy). Consequently, DOX often has a narrow therapeutic index and compromised therapeutic potential. DOX-induced myocardiopathy is a fatal ailment and with the development of heart failure the mortality rate is approximately 50%. Acute DOX cardiotoxicity is characterized by electrocardiogram irregularities like prolongation of the interval between the Q and T waves. With the absence of effective treatment for DOX-induced myocardiopathy, preventive measures include limiting the lifetime cumulative dose of Basic Live/dead differentiation Sensitivity Sensitivity 80% Cohort n = 20 60% Cohort n = 20 100% (S + C+) n = 11 82% (S + C+) n = 11 56% (S-C+) n = 9 33% (S-C+) n = 9 Specificity Specificity 84% Cohort n = 68 88% Cohort n = 68 87% (S–C–) n = 60 90% (S–C–) n = 63% (S + C-) n = 8 75% (S + C–) n = 8 Abstracts 3 DOX to b450 mg/m2 , use of continuous slow infusion in place of normal infusion protocols, replacement of DOX with anthracycline analogues, and use of different drug delivery carriers. The aim of this project is to develop a functionalized Pheroid® drug delivery carrier, with extravesicular targeting ligands specifically directing delivery of DOX to breast cancer cells which will increase therapeutic efficacy and reduce toxicity to healthy cells. In vitro patch clamp assays will be conducted to evaluate the effect of entrapping DOX in Pheroid® formulations on acute QT prolongation. We would analyze the IKr (rapid component of the delayed rectifier current) and IKs (second component of the delayed rectifier current) of cultured HEK (human embryonic kidney)-293 cells permanently transfected with the human ether-a-go-go related gene (hERG) and KCNQ1/KCNE1 genes, respectively, using the whole-cell configuration of the patch clamp technique. Functionalized Pheroids, with ligands specific to receptors overexpressed on breast cancer cells, as drug delivery vehicles for DOX could improve drug accumulation at the cancerous tissue through passive and active targeting and minimize acute cardiotoxicity of DOX. From this study we would be able to determine in vitro the preclinical safety with respect to the torsadogenic potential of formulations of DOX in Pheroids

The Pheroid® technology is a patented colloidal drug delivery system comprised of a dispersed oil phase of omega 3 and omega 6 fatty acids emulsified in N2O saturated water. Pheroid® has pharmaceutical applicability for the enhancement of therapeutic efficacy due to better drug exposure, reduced cytotoxicity and faster onset of action. The purpose of this paper was to evaluate the possible cardiovascular effects of the Pheroid® drug delivery system in the conscious rat. Six male rats instrumented with telemetry transmitters were used in a three-way crossover design. The effects of single intravenous administrations of Pheroid® and oral administration of Pheroid® and pro-Pheroid® on cardiovascular parameters were measured. These parameters included arterial blood pressure, heart rate and the ECG. As positive controls, a single intraperitoneal dose of clonidine (0.03 mg/kg) and an oral dose of theophylline (30 mg/kg) were also tested. No significant effects of Pheroid® on cardiovascular parameters were observed at any time following oral or intravenous administration. The expected effects of clonidine (decreased heart rate) and theophylline (increased heart rate) could be demonstrating confirming the sensitivity of the experimental model to detect changes in the cardiovascular parameters measured. We have demonstrated that Pheroid® has no effects on cardiovascular function following both oral and intravenous administration. These data showing cardiovascular safety are supportive for the use of Pheroid® for various pharmacological applications

Goserelin is a peptide drug that suppresses levels of testosterone and oestradiol [1]. The current formulation is presented in a sustained-release depot with several approved indications such as treatment of endometriosis, prostate cancer and breast cancer. The depots are placed subcutaneously with a large bore needle, hence there is a need for less invasive administration. An oral formulation in a drug delivery system such as Pheroid® would offer ease of administration to the patient, improved bioavailability as well as reduced manufacturing costs [2]. Therefore, the aim of this study was to determine the effect of entrapment of goserelin in Pheroid® on the pharmacokinetics (PK) of goserelin as well as the pharmacodynamic (PD) effects, in mice. Goserelin was entrapped in Pheroid® and characte- rized in terms of size, zeta potential and morphology. Male Balb/c mice, 6–8 weeks old were administered goserelin subcutaneously and goserelin entrapped in Pheroid® orally. At selected time points, mice were euthanised, blood collected and analysed for goserelin and testosterone levels by LCMS/MS to determine the PK and PD effects respectively. The size and zeta potential of Pheroid® remained in the expected ranges suggesting that entrapment of goserelin had no effect on Pheroid® vesicle size and stability. Confocal imaging showed satisfactory quality of the Pheroid® and goserelin did not have any effect on the Pheroid® structure. Entrapment of goserelin in Pheroid led to detectable levels of goserelin in plasma of the mice and a corresponding decrease in testosterone levels. The results suggest that Pheroid® may improve the oral bioavailability of goserelin and could be a potential strategy for an oral formulation which would be less invasive for patients

In general, N80% of the world's population relies on traditional medicine for their primary healthcare [1]. The use of complementary herbal medicine provides an alternative option to modern allopathic treatment for treating or preventing numerous diseases including cancer. According to the literature [2], a combination of two or more active biological compounds obtained from medicinal plants has a more profound therapeutic action than a single agent treatment. However, there is a paucity of information regarding the translation of phytochemicals to a potential phytopharmaceuticals. In addition, poor biological uptake coupled with inadequate safety and efficacy data hamper the clinical utility of most herbal medicines. This underscores the need for a thorough pragmatic study on the pharmacological efficacy and safety of herbal medicines including assessing the cardiovascular, neurological, hematological, respiratory, liver and kidney toxicities. Table 1 shows some examples of biologically active compounds derived from different medicinal plants and their toxicity. In the current study, the effective dose combination of three plant bioactive compounds, will be assessed in an in vitro assay against the human alveolar adenocarcinoma (A549) cell line. The biological uptake of the phytochemical combination will be enhanced using a novel Pheroid® drug delivery system. The in vivo anticancer activity of combined phytochemical Pheroid® formulation [3], as well as their impact on the peripheral blood parameters, and on liver and kidney functions will be evaluated for possible toxicity. See Table 1 for therapeutic activity and toxicity of herbal medicines [1]. It is anticipated that Pheroid® formulated phytochemical combinations will exhibit profound cytotoxic activity against the A549 lung cancer cells with minimal impact on the homeostasis of the body. Although herbal medicines are widely used as promising therapeutic agents, it is imperative to determine their efficacy, and assess potential adverse reactions for safe and rational use

The use of herbal medicine for treating various ailments has a deep history in South African traditional medicine. South Africa has a rich floral biodiversity which has been exploited for centuries mainly for day-to-day healthcare needs and food security. Almost 70% of South Africans rely on traditional medicine (TM) for their primary healthcare needs. There is a misconception that “natural” means safe from toxicity. However, a number of herbal medicines have been reported for severe side effects such as hepatotoxicity, renal failure and allergic reactions. The safety of TM is now a major concern for both health authorities and the general public. Safety pharmacology of herbal medicines is intended to detect potential adverse functional effects on various organ systems. The proposed criteria of safety pharmacology for herbal medicine should be the same as for Western drugs. The 10 most frequently used South African medicinal plants will be selected for this study. The Irwin test will be conducted to detect any untoward toxicity using Sprague-Dawley rats (n = 3). Aqueous extracts of the selected plants will be administered to the animals by oral gavage and the control group will receive solvent blank. The administered doses will exceed the therapeutic range just enough to produce moderate adverse effect. Telemetered rats will be used to assess the effects the selected herbal medicines will have on the core battery such as the cardiovascular, respiratory and CNS systems. All the experiments will follow the ARRIVE guidelines. Behaviours and clinical signals observed in the Irwin test, as well as data from telemetered rats will be used to identify the principle effects of all the test substances. South Africa is at the verge of incorporating traditional medicine into the mainstream healthcare system, robust safety pharmacology studies will ensure the safe of use of herbal medicine

Goserelin is a type of hormone therapy used in addition to standard adjuvant therapy. Currently, goserelin is only available as a slow releasing subcutaneous implant but this is an invasive and generally not well- tolerated method of administration. By using a drug delivery system (DDS), peptides can be protected from the harsh environment of the gastrointestinal (GI) tract. Pheroid® technology is one such example of a drug delivery system. This delivery system is a patented, colloidal drug delivery system and is capable of targeting specific areas for treatment, transportation of genetic material to the cellular nucleus and the decrease of drug resistance, all this while improving the delivery of dynamic complexes [1,2]. The prospect of changing to a less invasive administration method will yield a great advantage. Goserelin was investigated in female Balb/c mice. Pheroid® technology was used as a drug delivery system (DDS) to compare the bioavailability of goserelin subcutaneous depot against a peroral formulation with goserelin pro-Pheroid® formulation. The mice were divided into three groups; group one received goserelin subcutaneously, group 2 received an oral dose of 4 mg/kg pro-Pheroid®-goserelin and group 3 being the control group which received only pro- Pheroid®. Vaginal cytology was evaluated to track the disruption of the oestrus cycle Vaginal cytology results indicated a disruption in the oestrus cycle in the subcutaneous groups and the pro- Pheroid®-goserelin oral group. No disruption in the cycle was observed for the pro-Pheroid® oral group. Entrapment in Pheroid® resulted in oral bioavailability of goserelin as indicated by the related effect on the oestrus cycle. No disruption of the cycle was observed in the pro-pheroid® oral group, thus, goserelin was successfully absorbed in the blood after the peroral administration with pro-Pheroid. In conclusion, Pheroid® technology seemed to protect goserelin from the harsh environment of the gastrointestinal tract and could thus be a helpful DDS

Pheroid® is a drug delivery system with possible use in the treatment of chronic diseases such as cancer and that enhances the efficacy of the treatment of chronic diseases [1]. Current cancer treatment is non-selective, has serious long-term side-effects and sometimes is life-threatening [2]. Certain drug delivery systems can be used to improve the safety, stability and efficiency of cancer therapy [3]. Various types of Pheroid® can be formulated and custom-made for different applications [1]. Pheroid® has the potential to be formulated to specifically target cancer cells and to improve the bio- availability and efficacy of anti-cancer drugs. Pheroid® will be formulated with folic acid and transferrin covalently attached to the fatty acids that will be incorporated in the Pheroid®. These ligands are specifically chosen as the receptors for these ligands are overexpressed in certain cancer types. Mass spectrometry and NMR spectrometry will be used to confirm attachment. Cisplatin will be used as active ingredient that will be encapsulated in the Pheroid®. These formulations will be characterized and stability tests will be performed to determine the stability of the formulations. In vitro study will be performed with breast (MCF7) and lung (A549) cancer cell lines and will include cytotoxicity assays and cellular uptake assays to confirm selectivity. In vivo efficacy of the formulations will be performed on xenografts models of MCF-7 and A549 cell lines. A pharmacokinetic study and a tissue bio-distribution study will be performed to determine the selectivity of the Pheroid® formulations against cancer cells. The following results are anticipated for this study: Successfully attachment of ligands to fatty acids incorporated in Pheroid®; Selective targeting of cancer cells by Pheroid®; Accumulation of Pheroid® in tumor environment; Improve efficacy and safety of cisplatin treatment. This study will contribute to knowledge in the cancer research field and selective targeting of cancer cells by Pheroid® drug delivery system. We anticipate to patent the targeting of this technology and the use of it in cancer therapy