Histidine and lysine residues and the activity of phospholipase A2 from the venom of Bitis gabonica.

Chemical modification of phospholipase A2 (phosphatide 2-acyl-hydrolase, EC 3.1.1.4) from the venom of gaboon adder (Bitis gabonica) showed that histidine and lysine residues are essential for enzyme activity. Treatment with p-bromophenacyl bromide or pyridoxal 5'-phosphate resulted in the specific covalent modification of one histidine or a total of one lysine residue per molecule of enzyme, respectively, with a concomitant loss of enzyme activity. Competitive protection against modification and inactivation was afforded by the presence of Ca2+ and/or micellar concentrations of substrate analogue, lysophosphatidylcholine. Neither modification caused any significant conformational change, as judged from circular dichroic properties. Amino acid analyses and the alignment of peptides from cyanogen bromide and proteolytic cleavage of modified enzyme preparations delineated His-45 as the only residue modified by p-bromophenacyl bromide. However, pyridoxal 5'-phosphate was shown to have reacted not with a single lysine but with four different ones (residues 11, 33, 58 and 111) in such a manner that an overall stoichiometry of one modified lysine residue/molecule enzyme resulted. Apparently, the essential function of lysine could be fulfilled by any one out of these four residues.     

The potential application of FT-Raman spectroscopy for the quantification and mapping of the steroidal glycoside P57 in Hoodia gordonii

Hoodia gordonii, with the perceived active ingredient P57 (a steroidal glycoside), is a succulent plant which has gained commercial popularity as an anti-obesity preparation. The content of P57 is used as an indication of the quality of the raw material. Traditionally, quantification of P57 is performed using liquid-chromatography coupled to mass spectrometry (LC–MS) which is expensive and laborious. Vibrational spectroscopy methods such as FT-Raman spectroscopy offer a simple, less expensive and rapid alternative. The potential of FT-Raman to quantify and identify the location of P57 in H. gordonii raw plant material was investigated. LC–MS was used to determine the concentration of P57 in 145 plant samples and the data was used to develop a calibration model with chemometric techniques based on the partial least squares projections to latent structures (PLS) algorithm. The performance of the calibration model was evaluated according to the root mean square error of prediction (RMSEP) and correlation coefficient (R²). Pre-processing with orthogonal signal correction (OSC) yielded a model which predicted P57 content based on the FT-Raman spectra with a correlation coefficient (R²) value of 0.9986 and an RMSEP of 0.004%. These results demonstrate that FT-Raman spectroscopy holds great potential to rapidly quantify P57 in H. gordonii raw material with high accuracy as an alternative to LC–MS analysis. In addition, the spatial distribution of P57 in a cross-section of an H. gordonii stem sample was demonstrated using FT-Raman mapping showing that P57 is concentrated throughout the cortex which was confirmed with LC–MS.     

Efficacy of cultural control measures against the banana weevil, Cosmopolites sordidus (Germar), in South Africa

The banana weevil, Cosmopolites sordidus, is the most important insect pest of banana and plantain in the world. Cultural control methods were investigated over 2 years in southern KwaZulu‐Natal, South Africa. Harvesting at ground level and dissection of remnants (treatment 1), and covering the base of the mat (entire plant consisting of several meristems) with soil and moving debris to the inter‐row (treatment 2), were compared to a positive control that involved treatment of plants with a registered pesticide (treatment 3), and a negative control that involved harvesting at 150 cm from the collar with no soil or sanitation amendments (treatment 4). Yield, weevil damage and pseudostem girth of plants were measured from August to November annually, while adult beetle densities were assessed over 4 weeks in October/November and April. Nematode samples were taken and analysed in October/November every year. Damage parameters included the coefficient of infestation, the percentage coefficient of infestation (PCI) at two intervals, the summed PCI value, the percentage cross‐sectional damage of the central cylinder and cortex, and the mean cross‐sectional damage percentage. A randomized block design with three replicates was used in the trial. The parameters were similar before the onset of the trial. Fruit yield and plant girth, corrected by nematode densities, were not significantly different in any treatment, nor were the nematodes controlled. Soil cover and recession of remnants was the only effective treatment, significantly reducing the CI, but not the adult density or the other damage parameters. Soil cover showed promise as a cultural control method because it only needs to be applied seasonally and reduced the percentage cross‐sectional damage of the central cylinder, the damage parameter most closely related to yield, by 14%.     

Simple 1,4-benzoquinones with antibacterial activity from stems and leaves of Gunnera perpensa

From the dichloromethane extract of the leaves and stems of Gunnera perpensa two new, simple 1,4-benzoquinones and a known benzopyran-6-ol were isolated. From the methanol extract phytol was obtained. The two benzoquinones, 2-methyl-6-(-3-methyl-2-butenyl)benzo-1,4-quinone (1) and 3-hydroxy-2-methyl-5-(3-methyl-2-butenyl)benzo-1,4-quinone (2) and the benzopyran, 6-hydroxy-8-methyl-2,2-dimethyl-2H-benzopyran (3) were examined for antimicrobial properties together with the crude stem, leaf and root extracts. Minimum inhibitory concentration (MIC) assays were used to quantify antimicrobial activity and the MIC values for the crude extracts of stems, roots and leaves ranged between 100 microg and >16 mg/ml against the eight microorganisms investigated. Compound 1 showed significant antimicrobial activity with the most sensitive organism being Staphylococcus epidermidis with an MIC of 9.8 microg/ml. For compound 2, no activity was noted. Compound 3 exhibited good activity against the yeasts Cryptococcus neoformans (75 microg/ml) and Candida albicans (37.5 microg/ml).     

Identification of major metabolites in Aloe littoralis by high-performance liquid chromatography-nuclear magnetic resonance spectroscopy

Examination of the leaf exudate of the South African species Aloe littoralis by reversed-phase HPLC revealed the presence of two major metabolites. The identification of the two compounds without isolation was attempted by HPLC-NMR based on separation using a C18 column eluting with a deuterium oxide:acetonitrile solvent gradient and an inverse HPLC-NMR probe. For each compound, one-dimensional proton spectra, and two-dimensional homonuclear COSY and TOCSY, and heteronuclear HSQC and HMBC, spectra were collected. On the basis of the data obtained, the metabolites were characterised as 10-hydroxyaloin B and deacetyllittoraloin.     

Prevalence of bovine tuberculosis in African buffalo at Kruger National Park

Bovine tuberculosis (BTB) was first detected in Kruger National Park (KNP) in a single African buffalo (Syncerus caffer) in 1990. In 1991/1992, 2,071 African buffalo were examined for BTB as part of a culling program that removed animals from all known herds in KNP. The prevalence of BTB in 1991/1992 was estimated to be 0%, 4.4% (+/-0.6%), and 27.1% (+/-1.4%), in the north, central, and south zones of KNP, respectively. In 1998, a stratified, two-stage cluster sampling method was used to estimate that the prevalence of BTB was 1.5% (+/-2.5%), 16% (+/-5.3%), and 38.2% (+/-6.3%), in the north, central, and south zones, respectively. This represented a significant increase in prevalence (P < or = 0.05) in the south and central zones, but not in the north zone. Continued monitoring of BTB in KNP is important for understanding disease transmission risks, potential population effects, and the efficacy of disease management strategies. The methodology and sample sizes used in 1998 are appropriate for future BTB monitoring in KNP.     

Chromones and anthrones from Aloe marlothii and Aloe rupestris

A phytochemical investigation of the leaf exudate of Aloe marlothii has resulted in the isolation of a new chromone (7-O-methylaloeresin A) and a new anthrone (5-hydroxyaloin A 6'-O-acetate). Furthermore 7-O-methylaloesin was isolated as a natural product for the first time from the leaf exudate of Aloe rupestris. The structure elucidation of these compounds was based on spectral data including 2D NMR. The chemotaxonomic value of 7-O-methylaloesin in Aloe series Asperifoliae and section Pachydendron is discussed.     

<Emphasis Type="Italic">R</Emphasis><Subscript>0</Subscript>: Host Longevity Matters

The basic reproduction ratio, R₀, is a fundamental concept in epidemiology. It is defined as the total number of secondary infections brought on by a single primary infection, in a totally susceptible population. The value of R₀ indicates whether a starting epidemic reaches a considerable part of the population and causes a lot of damage, or whether it remains restricted to a relatively small number of individuals. To calculate R₀ one has to evaluate an integral that ranges over the duration of the infection of the host. This duration is, of course, limited by remaining host longevity. So, R₀ depends on remaining host longevity and in this paper we show that for long-lived hosts this aspect may not be ignored for long-lasting infections. We investigate in particular how this epidemiological measure of pathogen fitness depends on host longevity. For our analyses we adopt and combine a generic within- and between-host model from the literature. To find the optimal strategy for a pathogen from an evolutionary point of view, we focus on the indicator \(R_0^{{opt}}\), i.e., the optimum of R₀ as a function of its replication and mutation rates. These are the within-host parameters that the pathogen has at its disposal to optimize its strategy. We show that \(R_0^{{opt}}\) is highly influenced by remaining host longevity in combination with the contact rate between hosts in a susceptible population. In addition, these two parameters determine whether a killer-like or a milker-like strategy is optimal for a given pathogen. In the killer-like strategy the pathogen has a high rate of reproduction within the host in a short time span causing a relatively short disease, whereas in the milker-like strategy the pathogen multiplies relatively slowly, producing a continuous small amount of offspring over time with a small effect on host health. The present research allows for the determination of a bifurcation line in the plane of host longevity versus contact rate that forms the boundary between the milker-like and killer-like regions. This plot shows that for short remaining host longevities the killer-like strategy is optimal, whereas for very long remaining host longevities the milker-like strategy is advantageous. For in-between values of host longevity, the contact rate determines which of both strategies is optimal.     

The Role of Glutamine Oxoglutarate Aminotransferase and Glutamate Dehydrogenase in Nitrogen Metabolism in Mycobacterium bovis BCG

Recent evidence suggests that the regulation of intracellular glutamate levels could play an important role in the ability of pathogenic slow-growing mycobacteria to grow in vivo. However, little is known about the in vitro requirement for the enzymes which catalyse glutamate production and degradation in the slow-growing mycobacteria, namely; glutamine oxoglutarate aminotransferase (GOGAT) and glutamate dehydrogenase (GDH), respectively. We report that allelic replacement of the Mycobacterium bovis BCG gltBD-operon encoding for the large (gltB) and small (gltD) subunits of GOGAT with a hygromycin resistance cassette resulted in glutamate auxotrophy and that deletion of the GDH encoding-gene (gdh) led to a marked growth deficiency in the presence of L-glutamate as a sole nitrogen source as well as reduction in growth when cultured in an excess of L-asparagine.