Habitat use and densities of co‐existing migrant Willow Warblers Phylloscopus trochilus and resident eremomelas Eremomela spp. in Zimbabwe

Capsule Migrant Willow Warblers occupy more woodland types and occur at higher densities than ecologically‐similar resident Afrotropical warblers.

Aims To compare population densities of Willow Warblers and eremomelas in adjacent acacia, mopane and miombo woodlands, and assess the abundance of potential invertebrate prey in each habitat type, in order to investigate whether Palearctic migrants use more open habitats and are more flexible in habitat use than their Afrotropical counterparts in the same feeding guild.

Methods Using distance sampling we carried out four replicated sets of point counts in acacia woodland and three sets of counts in miombo and mopane between December 1999 and February 2000. We noted the tree species in which we saw warblers foraging and took beating‐tray samples of potential arthropod prey present on tree foliage in each of the three habitats.

Results Willow Warbler density in acacia woodland increased from 1.80 ± 0.54 (se) birds/ha in early December to 7.15 ± 1.41 birds/ha in late January after influxes of later arrivals. Densities of Willow Warblers in miombo and mopane were much lower (1.14 ± 0.28 and 0.38 ± 0.23 birds/ha, respectively) and did not show significant changes. Burnt‐necked Eremomelas averaged 0.74 ± 0.34 birds/ha in acacia woodland, and in miombo densities of Green‐capped and Yellow‐bellied Eremomelas were 0.23 ± 0.17 and 0.34 ± 0.26 birds/ha, respectively. Densities in mopane were too low to estimate reliably. Willow Warblers and Green‐capped Eremomelas showed some apparent preferences in tree species used for foraging but differences in tree use were not obviously related to the abundance of arthropod taxa present as potential prey.

Conclusion Willow Warblers occupied more habitats at greater density than similar Afrotropical warblers. They appear to favour acacia, but their settlement patterns and the reasons for disparities between densities of immigrants and residents are unclear.     

Population estimates,trends and habitat associations of breeding Lapwing Vanellus vanellus,Curlew Numenius arquata and Snipe Gallinago gallinago in Northern Ireland in 1999

Capsule Declines in the breeding populations of Snipe, Lapwing and Curlew were recorded between April and June 1999 and compared with previous estimates in 1987.

Aims To compare populations of non-coastal breeding waders between 1987 and 1999.

Methods In 106 2 km × 2 km square tetrads, observers recorded the number of breeding pairs of waders and habitat details on 1:10 000 scale maps.

Results A significant decline of c. 60% for Lapwing and Curlew, and a non-significant decline of 30% for Snipe was recorded over 12 years. Concentrations of these species were found in County Tyrone, but Counties Antrim, Down and Armagh supported few breeding pairs of any species. Very few pairs of any species were recorded on improved grassland despite its widespread availability.

Conclusion A successful conservation strategy for these species must address the wider countryside and not just key sites. Intensive pastoral farming in upland and lowland areas and activities such as drainage and peat extraction will further reduce the suitability of open habitats for these wader species.     

On approaches to the functional restoration of salivary glands damaged by radiation therapy for head and neck cancer,with a review of related aspects of salivary gland morphology and development

Radiation therapy for cancer of the head and neck can devastate the salivary glands and partially devitalize the mandible and maxilla. As a result, saliva production is drastically reduced and its quality adversely altered. Without diligent home and professional care, the teeth are subject to rapid destruction by caries, necessitating extractions with attendant high risk of necrosis of the supporting bone. Innovative techniques in delivery of radiation therapy and administration of drugs that selectively protect normal tissues can reduce significantly the radiation effects on salivary glands. Nonetheless, many patients still suffer severe oral dryness. I review here the functional morphology and development of salivary glands as these relate to approaches to preventing and restoring radiation-induced loss of salivary function. The acinar cells are responsible for most of the fluid and organic material in saliva, while the larger ducts influence the inorganic content. A central theme of this review is the extent to which the several types of epithelial cells in salivary glands may be pluripotential and the circumstances that may influence their ability to replace cells that have been lost or functionally inactivated due to the effects of radiation. The evidence suggests that the highly differentiated cells of the acini and large ducts of mature glands can replace themselves except when the respective pools of available cells are greatly diminished via apoptosis or necrosis owing to severely stressful events. Under the latter circumstances, relatively undifferentiated cells in the intercalated ducts proliferate and redifferentiate as may be required to replenish the depleted pools. It is likely that some, if not many, acinar cells may de-differentiate into intercalated duct-like cells and thus add to the pool of progenitor cells in such situations. If the stress is heavy doses of radiation, however, the result is not only the death of acinar cells, but also a marked decline in functional differentiation and proliferative capacity of all of the surviving cells, including those with progenitor capability. Restoration of gland function, therefore, seems to require increasing the secretory capacity of the surviving cells, or replacing the acinar cells and their progenitors either in the existing gland remnants or with artificial glands.     

Anticancer effects and lysosomal acidification in A549 cells by Astaxanthin from Haematococcus lacustris

Astaxanthin (AXN) is known to have health benefits by epidemiological studies. Therefore, it is of interest to assess the effect of AXN (derived from indigenous unicellular green alga Haematococcus lacustris) to modulate cell cycle arrest, lysosomal acidification and eventually apoptosis using in vitro in A549 lung cancer cells. Natural extracts of astaxanthin were obtained by standardized methods as reported earlier and characterized by standard HPLC and MS. Treatment of A549 cells with AXN (purified fraction) showed significant reduction in cell viability (about 50%) as compared to crude extract at 50µM concentration. Thus, we show the anticancer effects and lysosomal acidification in A549 cells by Astaxanthin from Haematococcus lacustris for further consideration. Together, our results demonstrated the anticancer potential of AXN from Haematococcus lacustris, which is found to be mediated via its ability to induce cell cycle arrest, lysosomal acidification and apoptotic induction.     

Self-association of the Lentivirus protein, Nef

Background

The HIV-1 pathogenic factor, Nef, is a multifunctional protein present in the cytosol and on membranes of infected cells. It has been proposed that a spatial and temporal regulation of the conformation of Nef sequentially matches Nef's multiple functions to the process of virion production. Further, it has been suggested that dimerization is required for multiple Nef activities. A dimerization interface has been proposed based on intermolecular contacts between Nefs within hexagonal Nef/FynSH3 crystals. The proposed dimerization interface consists of the hydrophobic B-helix and flanking salt bridges between R105 and D123. Here, we test whether Nef self-association is mediated by this interface and address the overall significance of oligomerization.

Results

By co-immunoprecipitation assays, we demonstrated that HIV-1Nef exists as monomers and oligomers with about half of the Nef protomers oligomerized. Nef oligomers were found to be present in the cytosol and on membranes. Removal of the myristate did not enhance the oligomerization of soluble Nef. Also, SIVNef oligomerizes despite lacking a dimerization interface functionally homologous to that proposed for HIV-1Nef. Moreover, HIV-1Nef and SIVNef form hetero-oligomers demonstrating the existence of homologous oligomerization interfaces that are distinct from that previously proposed (R105-D123). Intracellular cross-linking by formaldehyde confirmed that SF2Nef dimers are present in intact cells, but surprisingly self-association was dependent on R105, but not D123. SIV_MAC239Nef can be cross-linked at its only cysteine, C55, and SF2Nef is also cross-linked, but at C206 instead of C55, suggesting that Nefs exhibit multiple dimeric structures. ClusPro dimerization analysis of HIV-1Nef homodimers and HIV-1Nef/SIVNef heterodimers identified a new potential dimerization interface, including a dibasic motif at R105-R106 and a six amino acid hydrophobic surface.

Conclusions

We have demonstrated significant levels of intracellular Nef oligomers by immunoprecipitation from cellular extracts. However, our results are contrary to the identification of salt bridges between R105 and D123 as necessary for self-association. Importantly, binding between HIV-1Nef and SIVNef demonstrates evolutionary conservation and therefore significant function(s) for oligomerization. Based on modeling studies of Nef self-association, we propose a new dimerization interface. Finally, our findings support a stochastic model of Nef function with a dispersed intracellular distribution of Nef oligomers.     

Structural determinants of HscA peptide-binding specificity

The Hsp70-class molecular chaperone HscA interacts specifically with a conserved (99)LPPVK(103) motif of the iron-sulfur cluster scaffold protein IscU. We used a cellulose-bound peptide array to perform single-site saturation substitution of peptide residues corresponding to Glu(98)-Ile(104) of IscU to determine positional amino acid requirements for recognition by HscA. Two mutant chaperone forms, HscA(F426A) with a DnaK-like arch structure and HscA(M433V) with a DnaK-like substrate-binding pocket, were also studied. Wild-type HscA and HscA(F426A) exhibited a strict preference for proline in the central peptide position (ELPPVKI), whereas HscA(M433V) bound a peptide containing a Pro-->Leu substitution at this location (ELPLVKI). Contributions of Phe(426) and Met(433) to HscA peptide specificity were further tested in solution using a fluorescence-based peptide-binding assay. Bimane-labeled HscA and HscA(F426A) bound ELPPVKI peptides with higher affinity than leucine-substituted peptides, whereas HscA(M433V) favored binding of ELPLVKI peptides. Fluorescence-binding studies were also carried out with derivatives of the peptide NRLLLTG, a model substrate for DnaK. HscA and HscA(F426A) bound NRLLLTG peptides weakly, whereas HscA(M433V) bound NRLLLTG peptides with higher affinity than IscU-derived peptides ELPPVKI and ELPLVKI. These results suggest that the specificity of HscA for the LPPVK recognition sequence is determined in part by steric obstruction of the hydrophobic binding pocket by Met(433) and that substitution with the Val(433) sidechain imparts a broader, more DnaK-like, substrate recognition pattern.