Human genetic research: emerging trends in ethics

Genetic research has moved from Mendelian genetics to sequence maps to the study of natural human genetic variation at the level of the genome. This past decade of discovery has been accompanied by a shift in emphasis towards the ethical principles of reciprocity, mutuality, solidarity, citizenry and universality.     

Ring1b-mediated H2A ubiquitination associates with inactive X chromosomes and is involved in initiation of X inactivation

Histone modifications are thought to serve as epigenetic markers that mediate dynamic changes in chromatin structure and regulation of gene expression. As a model system for understanding epigenetic silencing, X chromosome inactivation has been previously linked to a number of histone modifications including methylation and hypoacetylation. In this study, we provide evidence that supports H2A ubiquitination as a novel epigenetic marker for the inactive X chromosome (Xi) and links H2A ubiquitination to initiation of X inactivation. We found that the H2A-K119 ubiquitin E3 ligase Ring1b, a Polycomb group protein, is enriched on Xi in female trophoblast stem (TS) cells as well as differentiating embryonic stem (ES) cells. Consistent with Ring1b mediating H2A ubiquitination, ubiquitinated H2A (ubH2A) is also enriched on the Xi of both TS and ES cells. We demonstrate that the enrichment of Ring1b and ubH2A on Xi is transient during TS and ES cell differentiation, suggesting that the Ring1b and ubH2A are involved in the initiation of both imprinted and random X inactivation. Furthermore, we showed that the association of Ring1b and ubH2A with Xi is mitotically stable in non-differentiated TS cells.     

MARK2/EMK1/Par-1Balpha phosphorylation of Rab11-family interacting protein 2 is necessary for the timely establishment of polarity in Madin-Darby canine kidney cells

Rab11a, myosin Vb, and the Rab11-family interacting protein 2 (FIP2) regulate plasma membrane recycling in epithelial cells. This study sought to characterize more fully Rab11-FIP2 function by identifying kinase activities modifying Rab11-FIP2. We have found that gastric microsomal membrane extracts phosphorylate Rab11-FIP2 on serine 227. We identified the kinase that phosphorylated Rab11-FIP2 as MARK2/EMK1/Par-1Balpha (MARK2), and recombinant MARK2 phosphorylated Rab11-FIP2 only on serine 227. We created stable Madin-Darby canine kidney (MDCK) cell lines expressing enhanced green fluorescent protein-Rab11-FIP2 wild type or a nonphosphorylatable mutant [Rab11-FIP2(S227A)]. Analysis of these cell lines demonstrates a new role for Rab11-FIP2 in addition to that in the plasma membrane recycling system. In calcium switch assays, cells expressing Rab11-FIP2(S227A) showed a defect in the timely reestablishment of p120-containing junctional complexes. However, Rab11-FIP2(S227A) did not affect localization with recycling system components or the normal function of apical recycling and transcytosis pathways. These results indicate that phosphorylation of Rab11-FIP2 on serine 227 by MARK2 regulates an alternative pathway modulating the establishment of epithelial polarity.     

CD8+ Foxp3+ regulatory T cells are induced during graft-versus-host disease and mitigate disease severity

Regulatory T cells (Tregs), in particular CD4(+) Foxp3(+) T cells, have been shown to play an important role in the maintenance of tolerance after allogeneic stem cell transplantation. In the current study, we have identified a population of CD8(+) Foxp3(+) T cells that are induced early during graft-versus-host disease (GVHD), constitute a significant percentage of the entire Treg population, and are present in all major GVHD target organs. These cells expressed many of the same cell surface molecules as found on CD4(+) Tregs and potently suppressed in vitro alloreactive T cell responses. Induction of these cells correlated positively with the degree of MHC disparity between donor and recipient and was significantly greater than that observed for CD4(+)-induced Tregs (iTregs) in nearly all tissue sites. Mice that lacked the ability to make both CD8(+) and CD4(+) iTregs had accelerated GVHD mortality compared with animals that were competent to make both iTreg populations. The absence of both iTreg populations was associated with significantly greater expansion of activated donor T cells and increased numbers of CD4(+) and CD8(+) T cells that secreted IFN-γ and IL-17. The presence of CD8(+) iTregs, however, was sufficient to prevent increased GVHD mortality in the complete absence of CD4(+) Tregs, indicating at least one functional iTreg population was sufficient to prevent an exacerbation in GVHD severity, and that CD8(+) iTregs could compensate for CD4(+) iTregs. These studies define a novel population of CD8(+) Tregs that play a role in mitigating the severity of GVHD after allogeneic stem cell transplantation.     

Tree species effects on coupled cycles of carbon, nitrogen, and acidity in mineral soils at a common garden experiment

Forest biogeochemical cycles are shaped by effects of dominant tree species on soils, but the underlying mechanisms are not well understood. We investigated effects of temperate tree species on interactions among carbon (C), nitrogen (N), and acidity in mineral soils from an experiment with replicated monocultures of 14 tree species. To identify how trees affected these soil properties, we evaluated correlations among species-level characteristics (e.g. nutrient concentrations in leaf litter, wood, and roots), stand-level properties (e.g. nutrient fluxes through leaf litterfall, nutrient pools in stemwood), and components of soil C, N, and cation cycles. Total extractable acidity (acidity_tot) was correlated positively with mineral soil C stocks (R ²  = 0.72, P < 0.001), such that a nearly two-fold increase in acidity_tot was associated with a more than two-fold increase of organic C. We attribute this correlation to effects of tree species on soil acidification and subsequent mineral weathering reactions, which make hydrolyzing cations available for stabilization of soil organic matter. The effects of tree species on soil acidity were better understood by measuring multiple components of soil acidity, including pH, the abundance of hydrolyzing cations in soil solutions and on cation exchange sites, and acidity_tot. Soil pH and acidity_tot were correlated with proton-producing components of the soil N cycle (e.g. nitrification), which were positively correlated with species-level variability in fine root N concentrations. Soluble components of soil acidity, such as aluminum in saturated paste extracts, were more strongly related to plant traits associated with calcium cycling, including leaf and root calcium concentrations. Our results suggest conceptual models of plant impacts on soil biogeochemistry should be revised to account for underappreciated plant traits and biogeochemical processes.     

Lithologic controls on biogenic silica cycling in South African savanna ecosystems

The efficacy of higher plants at mining Si from primary and secondary minerals in terrestrial ecosystems is now recognized as an important weathering mechanism. Grassland ecosystems are a particularly large reservoir of biogenic silica and are thus likely to be a key regulator of Si mobilization. Herein, we examine the effects of parent material (basaltic and granitic rocks) on the range and variability of biogenic silica pools in grass-dominated ecosystems along two precipitation gradients of Kruger National Park, South Africa. Four soil pedons and adjacent dominant plant species were characterized for biogenic silica content. Our results indicate that although soils derived from basalt had less total Si and dissolved Si than soils derived from granite, a greater proportion of the total Si was made up of biogenically derived silica. In general, plants and soils overlying basaltic versus granitic parent material stored greater quantities of biogenic silica and had longer turnover times of the biogenic silica pool in soils. Additionally, the relative abundance of biogenic silica was greater at the drier sites along the precipitation gradient regardless of parent material. These results suggest that the biogeochemical cycling of Si is strongly influenced by parent material and the hydrologic controls parent material imparts on soils. While soils derived from both basalt and granite are strongly regulated by biologic uptake, the former is a “tighter” system with less loss of Si than the latter which, although more dependent on biogenic silica dissolution, has greater losses of total Si. Lithologic discontinuities span beyond grasslands and are predicted to also influence biogenic silica cycling in other ecosystems.     

Arv1 lipid transporter function is conserved between pathogenic and nonpathogenic fungi

The lipid transporter Arv1 regulates sterol trafficking, and glycosylphosphatidylinositol and sphingolipid biosyntheses in Saccharomyces cerevisiae. ScArv1 contains an Arv1 homology domain (AHD) that is conserved at the amino acid level in the pathogenic fungal species, Candida albicans and Candida glabrata. Here we show S. cerevisiae cells lacking Arv1 are highly susceptible to antifungal drugs. In the presence of drug, Scarv1 cells are unable to induce ERG gene expression, have an altered pleiotrophic drug response, and are defective in multi-drug resistance efflux pump expression. All phenotypes are remediated by ectopic expression of CaARV1 or CgARV1. The AHDs of these pathogenic fungi are required for specific drug tolerance, demonstrating conservation of function. In order to understand how Arv1 regulates antifungal susceptibility, we examined sterol trafficking. CaARV1/CgARV1 expression suppressed the sterol trafficking defect of Scarv1 cells. Finally, we show that C. albicansarv1/arv1 cells are avirulent using a BALB/c disseminated mouse model. We suggest that overall cell survival in response to antifungal treatment requires the lipid transporter function of Arv1.     

Climate change and local public health in the United States: preparedness, programs and perceptions of local public health department directors

While climate change is inherently a global problem, its public health impacts will be experienced most acutely at the local and regional level, with some jurisdictions likely to be more burdened than others. The public health infrastructure in the U.S. is organized largely as an interlocking set of public agencies at the federal, state and local level, with lead responsibility for each city or county often residing at the local level. To understand how directors of local public health departments view and are responding to climate change as a public health issue, we conducted a telephone survey with 133 randomly selected local health department directors, representing a 61% response rate. A majority of respondents perceived climate change to be a problem in their jurisdiction, a problem they viewed as likely to become more common or severe over the next 20 years. Only a small minority of respondents, however, had yet made climate change adaptation or prevention a top priority for their health department. This discrepancy between problem recognition and programmatic responses may be due, in part, to several factors: most respondents felt personnel in their health department--and other key stakeholders in their community--had a lack of knowledge about climate change; relatively few respondents felt their own health department, their state health department, or the Centers for Disease Control and Prevention had the necessary expertise to help them create an effective mitigation or adaptation plan for their jurisdiction; and most respondents felt that their health department needed additional funding, staff and staff training to respond effectively to climate change. These data make clear that climate change adaptation and prevention are not currently major activities at most health departments, and that most, if not all, local health departments will require assistance in making this transition. We conclude by making the case that, through their words and actions, local health departments and their staff can and should play a role in alerting members of their community about the prospect of public health impacts from climate change in their jurisdiction.