Neutrophil function in whole blood and after purification: Changes in receptor expression,oxidase activity and responsiveness to cytokines

Neutrophil function and plasma membrane receptor expression was measured in cell suspensions isolated by two separate procedures and in unfractionated whole blood. When cells were prepared by a combined dextran/ficoll procedure, their ability to generate reactive oxidants in response to fMet-Leu-Phe was greater than in corresponding cells isolated by a one-step procedure on Mono-Poly Resolving Medium (M-PRM). Cells prepared by both methods could be primedin vitro by rGM-CSF, but the priming ratio was greater in cells prepared by the latter method. The ability of neutrophils in whole blood to generate reactive oxidants in response to fMet-Leu-Phe was extremely low, but this was increased by more than 10 fold if the blood was pre-incubated with rGM-CSF. Similarly, expression of CD 11b and CD 16 was very low (or undetectable) in neutrophils in whole blood, but this was rapidly increased upon priming. Activation by PMA resulted in a down regulation of CD 16 expression as the receptor was shed from the cell surface. Neutrophils isolated by either the dextran/ficoll or the M-PRM method showed increased expression of receptors compared with those in whole blood, although this expression was lower in cells isolated by the latter method. These data indicate that the isolation procedures used to obtain purified neutrophils prime both receptor expression and oxidase function, although these effects are minimalised in isolation procedures using M-PRM. Furthermore, as CD 16 expression on neutrophils in whole blood is rapidly up-regulated during priming, it seems likely that, as for complement receptors, rapidly-mobilisable intracellular stores of this receptor exist.     

Mammalian sex chromosomes: Evolution of organization and function

Comparisons of chromosome size, morphology and gene arrangements between mammals of different species permit us to deduce the genome characteristics of the common ancestor, and to chart the changes that have occurred during the divergence of the two lineages. The more distantly related are the species compared, the more remote the common ancestor whose characteristics can be deduced. This means that, providing there are sufficient similarities to warrant comparison, the more divergent the species compared, the more significant the contribution to our understanding of the organization of an ancestral mammalian genome and the process of mammalian genome evolution. One of the genetic surprises of the last decade was the discovery that, although gross karyotypes of distantly related orders of eutherian mammals (e.g. cat, cow, rabbit, man) have diverged extensively, gene mapping studies reveal the presence of large chromosome segments conserved across at least 60 million years (O'Brien et al. 1988). This finding makes it worthwhile to extend genetic comparisons to the two groups of mammals most distantly related to eutherian mammals--marsupials and monotremes. Here we will review comparisons of the sex chromosomes in these three major groups of extant mammals, and show how they have led us to a new view of the evolution of mammalian sex chromosome organization and function in sex determination and X chromosome inactivation.     

Analysis of molting and metamorphosis in the ecdysteroid-deficient mutant L(3)3DTS of Drosophila melanogaster

The dominant temperature-sensitive mutation L(3)3^DTS (DTS-3) in Drosophila melanogaster causes lethality of heterozygotes during the third larval instar at the restrictive temperature (29°C). Temperature-shift experiments revealed two distinct temperature-sensitive periods, with lethal phases during the third larval instar (which may persist for 4 weeks) and during the late pupal stage. At 29°C mutant imaginal discs are unable to evert in situ, but did evert normally if cultured in the presence of exogenous ecdysterone or when implanted into wild-type larval hosts. The only morphologically abnormal tissue present in the lethal larvae is the ring gland, the prothoracic gland being greatly hypertrophied in third instar DTS-3 larvae. Injection of a single wild-type ring gland rescued these mutant larvae, indicating that the mutant gland is functionally, as well as morphologically, abnormal. Finally, the mutant larvae were shown to have less than 10% of the wild-type ecdysteroid levels. These results are all consistent with a proposed lesion in ecdysteroid hormone production in DTS-3 larvae. A comparison with the phenotypes of other “ecdysone-less” mutants is presented.     

Family ideation,immigration, and the racial state: explaining divergent family reunification policies in Britain and the US

Why do family reunification policies differ across liberal democracies? Established literature explains differentiation through reference to the logics of distinct “national models” of immigration policy-making. Drawing on critical race and feminist scholarship, this paper finds consistent racial logics in the political histories of family reunification policies in Britain and the US during the mid-twentieth century. In a context where the geopolitical power of each country was conditioned by an assertive antiracist internationalism, “family” provided a colourblind, ideational platform to rearticulate the state-based racial project of white supremacy. Despite an original convergence, family reunification diverged because of the context-specific racialization of the immigrant family in each case. In rooting divergent reunification policies in the logics of the racial state, this paper contributes to efforts to incorporate race into the sociology of immigration. The paper also develops recent feminist scholarship on the role of “family ideation” in building liberal-democratic immigration systems.     

Mechanism of APTX nicked DNA sensing and pleiotropic inactivation in neurodegenerative disease

The failure of DNA ligases to complete their catalytic reactions generates cytotoxic adenylated DNA strand breaks. The APTX RNA‐DNA deadenylase protects genome integrity and corrects abortive DNA ligation arising during ribonucleotide excision repair and base excision DNA repair, and APTX human mutations cause the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1). How APTX senses cognate DNA nicks and is inactivated in AOA1 remains incompletely defined. Here, we report X‐ray structures of APTX engaging nicked RNA‐DNA substrates that provide direct evidence for a wedge‐pivot‐cut strategy for 5′‐AMP resolution shared with the alternate 5′‐AMP processing enzymes POLβ and FEN1. Our results uncover a DNA‐induced fit mechanism regulating APTX active site loop conformations and assembly of a catalytically competent active center. Further, based on comprehensive biochemical, X‐ray and solution NMR results, we define a complex hierarchy for the differential impacts of the AOA1 mutational spectrum on APTX structure and activity. Sixteen AOA1 variants impact APTX protein stability, one mutation directly alters deadenylation reaction chemistry, and a dominant AOA1 variant unexpectedly allosterically modulates APTX active site conformations.     

Plant controls on Late Quaternary whole ecosystem structure and function

Plants and animals influence biomass production and nutrient cycling in terrestrial ecosystems; however, their relative importance remains unclear. We assessed the extent to which mega‐herbivore species controlled plant community composition and nutrient cycling, relative to other factors during and after the Late Quaternary extinction event in Britain and Ireland, when two‐thirds of the region's mega‐herbivore species went extinct. Warmer temperatures, plant–soil and plant–plant interactions, and reduced burning contributed to the expansion of woody plants and declining nitrogen availability in our five study ecosystems. Shrub biomass was consistently one of the strongest predictors of ecosystem change, equalling or exceeding the effects of other biotic and abiotic factors. In contrast, there was relatively little evidence for mega‐herbivore control on plant community composition and nitrogen availability. The ability of plants to determine the fate of terrestrial ecosystems during periods of global environmental change may therefore be greater than previously thought.     

Using human case studies to teach evolution in high school A.P. biology classrooms

Background

While recent research indicates that using human examples can be an engaging way to teach core evolutionary concepts such as natural selection and phylogenetic thinking, teachers still face potential conflicts and challenges that arise from cultural barriers to teaching and learning about evolution. The “Teaching Evolution through Human Examples” (TEtHE) project developed (1) a set of four curriculum mini-units for advanced placement (A.P.) biology that use human examples to teach evolutionary principles (Adaptation to Altitude, Evolution of Human Skin Color, Malaria, and What Does It Mean To Be Human?), and (2) a cultural and religious sensitivity (CRS) teaching strategies resource that includes background materials and two in-class activities to help teachers create a classroom environment to increase student willingness to engage the topic.

Methods

This paper reports on the development and field test of the TEtHE materials in A.P. biology classes in 10 schools in 8 states during the 2012–2013 school year using a design-based research framework (cf. Anderson and Shattuck in Educ Res 41:16–25, 2012). We chose A.P. classrooms to study the potential impacts of the materials in a “best case scenario” and analyzed data about understanding and acceptance of evolution from pre-post assessments in the 10 classrooms separately to mitigate potential validity concerns arising from the design (Anderson and Shattuck in Educ Res 41:16–25, 2012; Shadish et al. in Experimental and quasi-experimental designs for generalized causal inference. Houghton Mifflin, Boston, 2002). These data were treated as a secondary source of formative data to add additional perspective to teacher self-reports, observations, student and teacher questionnaires, teacher interviews, and student focus groups.

Results

Results indicate that the use of the three curriculum mini-units which focus on natural selection and the CRS classroom activities generally increased A.P. biology students’ understanding and acceptance of evolution. Students whose teachers used one of the CRS activities showed generally larger increases in understanding of evolution than those whose teachers did not use one of the CRS activities.

Conclusions

Although the utility of using human examples to teach evolution in college-level classes has been demonstrated in a few previous studies, this is the first national project of which we are aware to systematically explore the effect of a similar approach in high school biology classes. While we recognize that the results may be mitigated by the limitations of design-based research, particularly the absence of a comparison or control group, the general effectiveness of this approach suggested by qualitative and quantitative data in increasing student understanding and acceptance of evolution suggests that using human examples and explicitly creating a classroom environment to help students engage the topic of evolution are worth considering for further development and more robust testing.

     

mtDNA sequence diversity in Africa.

mtDNA sequences were determined from 241 individuals from nine ethnic groups in Africa. When they were compared with published data from other groups, it was found that the !Kung, Mbuti, and Biaka show on the order of 10 times more sequence differences between the three groups, as well as between those and the other groups (the Fulbe, Hausa, Tuareg, Songhai, Kanuri, Yoruba, Mandenka, Somali, Tukana, and Kikuyu), than these other groups do between one other. Furthermore, the pairwise sequence distributions, patterns of coalescence events, and numbers of variable positions relative to the mean sequence difference indicate that the former three groups have been of constant size over time, whereas the latter have expanded in size. We suggest that this reflects subsistence patterns in that the populations that have expanded in size are food producers whereas those that have not are hunters and gatherers.