Deforestation and the spatio-temporal distribution of savannah and forest members of the Simulium damnosum complex in southern Ghana and south-western Togo

Spatio-temporal data on cytotaxonomic identifications of larvae of different members of the Simulium damnosum complex collected from rivers in southern Ghana and south-western Togo from 1975 until 1997 were analysed. When the data were combined, the percentages of savannah blackflies (S. damnosum sensu stricto and S. sirbanum) in the samples were shown to have been progressively increasing since 1975. The increases were statistically significant (P < 0·001), but the rates of increase were not linear. Further analyses were conducted according to the collection seasons and locations of the samples, to account for possible biases such as savannah flies occurring further south in the dry season or a preponderance of later samples from northern rivers having more savannah flies. These analyses showed that the increasing trend was statistically significant (P< 0·0001) only during the periods April to June and October to December. The presence of adult savannah flies carrying infective larvae (L3) indistinguishable from those of Onchocerca volvulus in the study zone was confirmed by examinations of captured flies. The percentages of savannah flies amongst the human-biting populations and the percentages with L3s in the head were higher during dry seasons than wet seasons and the savannah species were found furthest south (5 °25′N) in the dry season. Comparisons of satellite images taken in 1973 and 1990 over a study area in south-western Ghana encompassing stretches of the Tano and Bia rivers demonstrated that there have been substantial increases in urban and savannah areas, at the expense of forest. This was so not only for the whole images but also for subsamples of the images taken at 1, 2, 4, 8 and 16 km distant from sites alongside the River Tano. At every distance from the river, the percentages of pixels classified as urban or savannah have increased in 1990 compared with 1973, while those classified as degraded or dense forest have decreased. The possibility that the proportionate increases in savannah forms of the vectors of onchocerciasis, and hence in the likelihood of the transmission of savannah strains of the disease in formerly forested areas, were related to the decreases in forest cover is discussed.

The use of fluorescence microscopy to define polymer localisation to the late endocytic compartments in cells that are targets for drug delivery

Macromolecular therapeutics and nano-sized drug delivery systems often require localisation to specific intracellular compartments. In particular, efficient endosomal escape, retrograde trafficking, or late endocytic/lysosomal activation are often prerequisites for pharmacological activity. The aim of this study was to define a fluorescence microscopy technique able to confirm the localisation of water-soluble polymeric carriers to late endocytic intracellular compartments. Three polymeric carriers of different molecular weight and character were studied: dextrin (Mw~50,000 g/mol), a N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer (Mw approximately 35,000 g/mol) and polyethylene glycol (PEG) (Mw 5000 g/mol). They were labelled with Oregon Green (OG) (0.3-3 wt.%; <3% free OG in respect of total). A panel of relevant target cells were used: THP-1, ARPE-19, and MCF-7 cells, and primary bovine chondrocytes (currently being used to evaluate novel polymer therapeutics) as well as NRK and Vero cells as reference controls. Specific intracellular compartments were marked using either endocytosed physiological standards, Marine Blue (MB) or Texas-red (TxR)-Wheat germ agglutinin (WGA), TxR-Bovine Serum Albumin (BSA), TxR-dextran, ricin holotoxin, C6-7-nitro-2,1,3-benzoxadiazol-4-yl (NBD)-labelled ceramide and TxR-shiga toxin B chain, or post-fixation immuno-staining for early endosomal antigen 1 (EEA1), lysosomal-associated membrane proteins (LAMP-1, Lgp-120 or CD63) or the Golgi marker GM130. Co-localisation with polymer-OG conjugates confirmed transfer to discreet, late endocytic (including lysosomal) compartments in all cells types. The technique described here is a particularly powerful tool as it circumvents fixation artefacts ensuring the retention of water-soluble polymers within the vesicles they occupy.

Polyunsaturated fatty acids in the pathogenesis and treatment of multiple sclerosis

Epidemiological, biochemical, animal model and clinical trial data described in this overview strongly suggest that polyunsaturated fatty acids, particularly n-6 fatty acids, have a role in the pathogenesis and treatment of multiple sclerosis (MS). Data presented provides further evidence for a disturbance in n-6 fatty acid metabolism in MS. Disturbance of n-6 fatty acid metabolism and dysregulation of cytokines are shown to be linked and a "proof of concept clinical trial" further supports such a hypothesis. In a randomised double-blind, placebo controlled trial of a high dose and low dose selected GLA (18:3n-6)-rich oil and placebo control, the high dose had a marked clinical effect in relapsing-remitting MS, significantly decreasing the relapse rate and the progression of disease. Laboratory findings paralleled clinical changes in the placebo group in that production of mononuclear cell pro-inflammatory cytokines (TNF-alpha, IL-1 beta) was increased and anti-inflammatory TGF-beta markedly decreased with loss of membrane n-6 fatty acids linoleic (18:2n-6) and arachidonic acids (20:4n-6). In contrast there were no such changes in the high dose group. The improvement in disability (Expanded Disability Status Scale) in the high dose suggests there maybe a beneficial effect on neuronal lipids and neural function in MS. Thus disturbed n-6 fatty acid metabolism in MS gives rise to loss of membrane long chain n-6 fatty acids and loss of the anti-inflammatory regulatory cytokine TGF-beta, particularly during the relapse phase, as well as loss of these important neural fatty acids for CNS structure and function and consequent long term neurological deficit in MS.