Replicating cells undergo DNA synthesis in the highly regulated, S-phase of the cell cycle. Analogues of the pyrimidine deoxynucleoside thymidine may be inserted into replicating DNA, effectively tagging dividing cells allowing their characterisation. Tritiated thymidine, targeted using autoradiography was technically demanding and superseded by 5-bromo-2-deoxyuridine (BrdU) and related halogenated analogues, detected using antibodies. Their detection required the denaturation of DNA, often constraining the outcome of investigations. Despite these limitations BrdU alone has been used to target newly synthesised DNA in over 20,000 reviewed biomedical studies. A recent breakthrough in “tagging DNA synthesis” is the thymidine analogue 5-ethynyl-2′-deoxyuridine (EdU). The alkyne group in EdU is readily detected using a fluorescent azide probe and copper catalysis using ‘Huisgen’s reaction’ (1,3-dipolar cycloaddition or ‘click chemistry’). This rapid, two-step biolabelling approach allows the tagging and imaging of DNA within cells whilst preserving the structural and molecular integrity of the cells. The bio-orthogonal detection of EdU allows its application in more experimental assays than previously possible with other “unnatural bases”. These include physiological, anatomical and molecular biological experimentation in multiple fields including, stem cell research, cancer biology, and parasitology. The full potential of EdU and related molecules in biomedical research remains to be explored.
The patterning of the Arabidopsis root epidermis depends on a genetic regulatory network that operates both within and between cells. Genetic studies have identified a number of key components of this network, but a clear picture of the functional logic of the network is lacking. Here, we integrate existing genetic and biochemical data in a mathematical model that allows us to explore both the sufficiency of known network interactions and the extent to which additional assumptions about the model can account for wild-type and mutant data. Our model shows that an existing hypothesis concerning the autoregulation of WEREWOLF does not account fully for the expression patterns of components of the network. We confirm the lack of WEREWOLF autoregulation experimentally in transgenic plants. Rather, our modelling suggests that patterning depends on the movement of the CAPRICE and GLABRA3 transcriptional regulators between epidermal cells. Our combined modelling and experimental studies show that WEREWOLF autoregulation does not contribute to the initial patterning of epidermal cell fates in the Arabidopsis seedling root. In contrast to a patterning mechanism relying on local activation, we propose a mechanism based on lateral inhibition with feedback. The active intercellular movements of proteins that are central to our model underlie a mechanism for pattern formation in planar groups of cells that is centred on the mutual support of two cell fates rather than on local activation and lateral inhibition.
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Helminths cause great economic loss in livestock in Africa, and can be categorized as either direct or indirect losses. Arid and semi-arid lands (ASAL) in Kenya comprise 71 % of total land area and harbour the largest population of cattle, sheep and goats. However, little information on the distribution and impact of gastro-intestinal (GIT) parasitism in these animals is available. This survey was conducted to establish the prevalence of GIT parasites infecting calves, sheep and goats and their relative importance in Magadi division, which is semi-arid. Faecal samples were obtained directly from the rectum of 109 calves, 133 goats and 20 sheep and submitted to the laboratory for faecal worm egg counts, and coccidial oocysts examination using a modified McMaster method. The significance of differences in mean egg count per gram (epg) between animal species and herds (farms) were assessed using analysis of variance. The overall prevalence of nematodes in the calves, sheep and goats was 69.2 %, 80 % and 82 %, respectively. About 10 % of sheep and goats had epgs higher than 1 000, the remainder having light to moderate infections. The overall prevalence of coccidial oocysts in calves, sheep and goats was 30 %, 44 % and 45 %, respectively. Poor productivity in ASAL areas, where nutrition is often poor, is likely to be pronounced in the presence of parasite infections. These findings indicate that viable internal parasite control should be implemented in the study area in order to increase the productivity of the livestock there.
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