Protein Identification of Venoms of the African Spitting Cobras, Naja mossambica and Naja nigricincta nigricincta.

Cobra snakes, including Naja mossambica and Naja nigricincta nigricincta, are one of the major groups of snakes responsible for snakebites in southern Africa, producing significant cytotoxicity and tissue damage. The venom of N. mossambica has been briefly characterised, but that of N. n. nigricincta is not reported. The current study identifies the venom proteins of N. mossambica and N. n. nigricincta. This is achieved using sodium dodecyl sulphate (SDS)-polyacrylamide gel eletrophroresis (PAGE), followed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Most of the proteins were less than 17 kDa in both snakes. N. mossambica was found to have 75 proteins in total (from 16 protein families), whereas N.n. nigricincta had 73 (from 16 protein families). Of these identified proteins, 57 were common in both snakes. The proteins identified belonged to various families, including the three-finger toxins (3FTx), Cysteine-rich secretory proteins (CRiSP), Phospholipase A2 (PLA2) and Venom metalloproteinase M12B (SVMP). The current study contributes to the profile knowledge of snake venom compositions, which is of fundamental value in understanding the proteins that play a major role in envenomation.

Structure-Activity Studies on the Hypertrehalosemic Hormone II of the Stick Insect Carausius morosus (Phasmatodea): Carbohydrate-Mobilization and Cardio-Stimulatory Activities.

The corpora cardiaca of the Indian stick insect, Carausius morosus, synthesize two decapeptide neuropeptides of the adipokinetic hormone (AKH) family, both of which can increase the trehalose levels in the hemolymph when the stick insect is ligated between the head and the thorax. Here, we use two biological assays to assess the potencies of 19 AKH analogs in ligated C. morosus: the carbohydrate-mobilizing assay measures the change in the levels of circulating carbohydrates following injection of a substance, while the semi-exposed heart assay measures a change in heart beat rate after the peptide is applied onto the heart. With the endogenous AKH (Carmo-HrTH-II) as lead peptide, we report here on seven naturally-occurring AKH peptides (bioanalogs) selected for testing because of a single or double amino acid replacement, or for being octapeptides. Single amino acid substitutions by an alanine residue at all positions of Carmo-HrTH-II, as well as analogs modified at the termini were also investigated to give a comprehensive view of ligand-receptor interaction at the physiological level in a hemimetabolous insect that practices thanatosis (feigning death). Only small changes are elicited in the bioassays, but the results from the two tests are comparable bar one or two anomalies. Results show that analogs modified at the termini have no or reduced activity. Regarding structural requirements of a ligand, the C. morosus AKH receptor appears to be strict: octapeptides are not preferred and many of the decapeptide analogs failed to reach 50% activity relative to Carmo-HrTH-II. The data implies that the AKH receptor in C. morosus mostly does not tolerate shorter peptides and single amino acid replacements in most places of the native AKH peptide. This information is important if environmentally friendly insect-specific pesticides are made based on an insect AKH as lead peptide: stick insects that are normally not viewed as pest insects may not be easily targeted by cross-reactive AKH mimetics directed at harmful insects, due to the very specific amino acid requirements to activate the C. morosus AKH receptor.

A pragmatic approach to introducing mixed methods research training for pharmacy students in Namibia.

BACKGROUND AND PURPOSE: Pharmacy and medical training were introduced for the first time in Namibia in 2011 and 2010 respectively. All students must complete a research project as part of their training, which is supported by various courses in the respective curricula including research methods. Following a revision of the medical curriculum, there was an opportunity to review the way research methods was taught for both degrees, piloting in pharmacy then expanding to other disciplines. An educational activity that was part of the research methods course for training pharmacy students in Namibia is described. EDUCATIONAL ACTIVITY AND SETTING: The activity described related to a new approach in the running of the research methods course for pharmacy students and included, in a portfolio-based approach, a group project. Students were tasked to collect qualitative data from medical student colleagues that they then needed to codify and ultimately articulate into a survey questionnaire. The questionnaire was subsequently sent out to collect responses on medicines-related items that could be analyzed quantitatively in subsequent teaching sessions. Supportive lectures, tutorials and portfolio assignments were provided during the project. Data were collected the following year to create a more substantive dataset and a screencast video made to benefit future students in the course. FINDINGS AND DISCUSSION: Through the course of this activity students developed a questionnaire survey tool based on qualitative responses to brief interviews with medical students and emergent themes based on qualitative analyses. A dataset was created that allowed demonstration of quantitative analyses and extraction of sub-scales from the questionnaire. Further educational resources were developed to ensure sustainability of this educational resource and retention in the taught curriculum. SUMMARY: The current article discusses the development, implementation and evaluation of this research methods course component. The application of data collected as part of the activity and its relevance to the educational activity is examined as well as lessons learned for the future running of the project and further evaluation.

Influence of aminergic and peptidergic substances on heart beat frequency in the stick insect Carausius morosus (Insecta, Phasmatodea).

The dorsal heart of the Indian stick insect, Carausius morosus, is responsible for the anterograde flow of hemolymph to the aorta and into the body cavity. The contraction frequency of the insect heart is known to be influenced by several substances of neural source. Here, a semi-exposed heart assay was employed to study the effect of an aminergic substance (octopamine) and three neuropeptides (C. morosus hypertrehalosemic hormone [Carmo-HrTH], crustacean cardioactive peptide [CCAP], and proctolin) on heart contraction. The contraction frequency was measured as beats per minute in adults ligated between the head and the prothorax. All three investigated neuropeptides had a stimulatory effect on heart contraction that lasted approximately 6 min, after which the normal heart beat rate was restored. Proctolin and CCAP stimulated the rate of heart beat also in unligated stick insects, whereas Carmo-HrTH was active only in ligated insects. The latter could suggest that when the stick insect is not ligated, a competing substance may be released from the head of C. morosus; the competing substance is, apparently, not physiologically active but it binds or blocks access to the receptor of Carmo-HrTH-II, thereby rendering the HrTH peptide "not active." In ligated stick insects, 6.7 × 10-8  M Carmo-HrTH-II significantly increased the heart beat rate; higher doses resulted in no further increase, suggesting the saturation of the HrTH receptor. Octopamine inhibited the rate at which the heart contracted in a dose-dependent manner; inhibition was achieved with 10-4  M of octopamine.