The AIDS of aid?: long-term organisation challenges of a CBO dealing with HIV/AIDS, poverty and donor aid

The following treatise first frames the role of CBOs in responding to the HIV/Aids crisis in relation to their position in the global health governance system through a literature survey that moves from an analysis of the global structures down to the local. The survey covers the role of international organisations, international NGOs (INGOs), national governments, local NGOs and CBOs and outlines the context in which Masizakhe is working within the global health governance structure. Secondly the research design and methodology are outlined focusing on the longitudinal, case study and participant--‐observation approaches. Hypotheses, conceptualisation, definitions, key variables are described and data collection methods and fieldwork practice extrapolated upon. Following that data capturing, editing and analysis are discussed in conjunction with shortcomings and sources of error. In the fourth chapter the research discusses the history, structure and outlines the research findings by comparing what has changed within the organisation over time, presenting and discussing the results. The outcomes of this research have shown that existing problems in this particular CBO are very difficult to overcome without committed, sustained support from donors, government, community and the organisation’s members. CBOs are often hamstrung by a series of intersecting factors which hamper their ability to problem--‐solve, even when the route to overcoming the problem is clear, particularly when the capacity and will to do so is not always present from both within the organisation and from outside support systems. These challenges then impact on the overall quality of and ability to deliver the services the organisation is structured to deliver. The major challenge for the organisation remains the inconsistent donor cycle and resultant instability thus created within an organisation already working in a highly volatile, unstable situation marked by poverty and disease. Thus the title, The Aids of Aid?, captures the essence of Masizakhe’s struggle with its own syndrome of problems. It summarises a comment made by the project secretary said that: “Sometimes it feels like we are not only fighting for the health of our people – We are fighting for the health of our organisation. We are a sick organisation trying to help sick people. All we need is donors and funding –we can’t live without them, and when they don't give, we get sick” (Stamper, Pers Comm, 2011). The other emergent challenges were a battle internally with ‘founder syndrome’, lack of management transparency and a dysfunctional board

Detection and quantification of residues in sheep exposed to trace levels of dietary zilpaterol HCl

Study objectives were to determine zilpaterol residues in urine and tissues of sheep fed dietary zilpaterol HCl, at levels commensurate with feed contamination, using common and novel screening and quantitative analytical methods. Sheep (50.0 ± 2.7 kg) were offered feed (1.75 kg/d) containing 0.0075 (L), 0.075 (M), or 0.75 (H) mg kg−1 of zilpaterol for 12 days and were slaughtered with 0-day (L-0, M-0, H-0; n = 4 each) or 3-day (H-3; n = 4) withdrawal periods. Rapid immunochromatographic assays (ICA) consistently detected urinary zilpaterol (LOD = 1.7 ng mL−1) in L-0 (54.2%), M-0 (96.0%), and the H-0 (100%) treatment groups but only detected zilpaterol in tissues (LOD ~2.4 ng g−1) from the H-0 group. Advanced MS-based technologies detected zilpaterol in some, but not all, tissues of M-0, H-0, L-0, and H-3 sheep. Analytical techniques commonly used to ensure compliance with show-animal rules, import/export guidelines, and regulatory statutes routinely detected residues in animals exposed to zilpaterol at doses insufficient to elicit growth response

Evaluation of penicillin G residues by kidney inhibition swab tests in sow body fluids and tissues following intramuscular injection

In 2011, the USDA-Food Safety and Inspection Service (FSIS) changed the method used for screening swine tissues for antimicrobial residues from the Fast Antimicrobial Screen Test to the Kidney Inhibition Swab (KIS™). Here, we describe the use of KIS™ test for the detection of penicillin G residues in kidney, liver, plasma, urine, and skeletal muscle of heavy sows following the administration of a 5x label dose of penicillin G procaine. Such off-label use is legal in the United States under the Animal Medicinal Drug Use Clarification Act (AMDUCA) when label routes or doses are ineffective at treating disease and is commonly used to treat bacterial infections in heavy sows

A validated HPLC method for the simultaneous determination of bleomycin A2 and B2 in human plasma

Bleomycin is an anti-neoplastic drug that has recently been used for the treatment of vascular anomalies. The plasma concentration of bleomycin following intralesional injection into vascular lesions is unknown. An expedient method was developed for the determination of plasma bleomycin levels using ion-paired reversed phase high performance liquid chromatography (HPLC). Prior to sample analysis using the HPLC, a one-step protein precipitation sample preparation procedure was used to remove proteins from the plasma. The calibration curve was linear over the range 0.1 - 0.8μg/mL bleomycin A2 and 0.2 – 1.0μg/mL bleomycin B2. Recovery was approximately 100%. This method provides a simple and rapid way of determining the levels of bleomycin A2 and B2 in patient plasma.Bristol-Myers Squibb and the National Research Foundationhttp://www.pharmscidirect.com/am2013ay201

The Structural Basis of Gas-Responsive Transcription by the Human Nuclear Hormone Receptor REV-ERBβ

Heme is a ligand for the human nuclear receptors (NR) REV-ERBα and REV-ERBβ, which are transcriptional repressors that play important roles in circadian rhythm, lipid and glucose metabolism, and diseases such as diabetes, atherosclerosis, inflammation, and cancer. Here we show that transcription repression mediated by heme-bound REV-ERBs is reversed by the addition of nitric oxide (NO), and that the heme and NO effects are mediated by the C-terminal ligand-binding domain (LBD). A 1.9 Å crystal structure of the REV-ERBβ LBD, in complex with the oxidized Fe(III) form of heme, shows that heme binds in a prototypical NR ligand-binding pocket, where the heme iron is coordinately bound by histidine 568 and cysteine 384. Under reducing conditions, spectroscopic studies of the heme-REV-ERBβ complex reveal that the Fe(II) form of the LBD transitions between penta-coordinated and hexa-coordinated structural states, neither of which possess the Cys384 bond observed in the oxidized state. In addition, the Fe(II) LBD is also able to bind either NO or CO, revealing a total of at least six structural states of the protein. The binding of known co-repressors is shown to be highly dependent upon these various liganded states. REV-ERBs are thus highly dynamic receptors that are responsive not only to heme, but also to redox and gas. Taken together, these findings suggest new mechanisms for the systemic coordination of molecular clocks and metabolism. They also raise the possibility for gas-based therapies for the many disorders associated with REV-ERB biological functions

Selection of Anti-Sulfadimidine Specific ScFvs from a Hybridoma Cell by Eukaryotic Ribosome Display

BACKGROUND:Ribosome display technology has provided an alternative platform technology for the development of novel low-cost antibody based on evaluating antibiotics derived residues in food matrixes. METHODOLOGY/PRINCIPAL FINDINGS:In our current studies, the single chain variable fragments (scFvs) were selected from hybridoma cell lines against sulfadimidine (SM(2)) by using a ribosome library technology. A DNA library of scFv antibody fragments was constructed for ribosome display, and then mRNA-ribosome-antibody (MRA) complexes were produced by a rabbit reticulocyte lysate system. The synthetic sulfadimidine-ovalbumin (SM(2)-OVA) was used as an antigen to pan MRA complexes and putative scFv-encoding genes were recovered by RT-PCR in situ following each panning. After four rounds of ribosome display, the expression vector pCANTAB5E containing the selected specific scFv DNA was constructed and transformed into Escherichia coli HB2151. Three positive clones (SAS14, SAS68 and SAS71) were screened from 100 clones and had higher antibody activity and specificity to SM(2) by indirect ELISA. The three specific soluble scFvs were identified to be the same molecular weight (approximately 30 kDa) by Western-blotting analysis using anti-E tag antibodies, but they had different amino acids sequence by sequence analysis. CONCLUSIONS/SIGNIFICANCE:The selection of anti-SM(2) specific scFv by in vitro ribosome display technology will have an important significance for the development of novel immunodetection strategies for residual veterinary drugs

The Cysteine Rich Necrotrophic Effector SnTox1 Produced by Stagonospora nodorum Triggers Susceptibility of Wheat Lines Harboring Snn1

The wheat pathogen Stagonospora nodorum produces multiple necrotrophic effectors (also called host-selective toxins) that promote disease by interacting with corresponding host sensitivity gene products. SnTox1 was the first necrotrophic effector identified in S. nodorum, and was shown to induce necrosis on wheat lines carrying Snn1. Here, we report the molecular cloning and validation of SnTox1 as well as the preliminary characterization of the mechanism underlying the SnTox1-Snn1 interaction which leads to susceptibility. SnTox1 was identified using bioinformatics tools and verified by heterologous expression in Pichia pastoris. SnTox1 encodes a 117 amino acid protein with the first 17 amino acids predicted as a signal peptide, and strikingly, the mature protein contains 16 cysteine residues, a common feature for some avirulence effectors. The transformation of SnTox1 into an avirulent S. nodorum isolate was sufficient to make the strain pathogenic. Additionally, the deletion of SnTox1 in virulent isolates rendered the SnTox1 mutated strains avirulent on the Snn1 differential wheat line. SnTox1 was present in 85% of a global collection of S. nodorum isolates. We identified a total of 11 protein isoforms and found evidence for strong diversifying selection operating on SnTox1. The SnTox1-Snn1 interaction results in an oxidative burst, DNA laddering, and pathogenesis related (PR) gene expression, all hallmarks of a defense response. In the absence of light, the development of SnTox1-induced necrosis and disease symptoms were completely blocked. By comparing the infection processes of a GFP-tagged avirulent isolate and the same isolate transformed with SnTox1, we conclude that SnTox1 may play a critical role during fungal penetration. This research further demonstrates that necrotrophic fungal pathogens utilize small effector proteins to exploit plant resistance pathways for their colonization, which provides important insights into the molecular basis of the wheat-S. nodorum interaction, an emerging model for necrotrophic pathosystems