The influence of Ca2+ on the lateral lipid distribution and phase transition in phosphatidylethanolamine/phosphatidylserine vesicles

The lateral lipid distribution within dipalmitoylphosphatidylethanolamine (DPPE)/dipalmitoylphosphatidylserine (DPPS) vesicle membranes was investigated under the influence of Ca²⁺ using a lipid cross-linking method. To characterize the phase transition in DPPE/DPPS vesicles and to correlate the different phase states of the membrane lipids with the obtained lipid distribution ESR measurements using a fatty acid spin label were carried out. It is shown that Ca²⁺ has a significant influence on the lateral lipid distribution within the fluid phase of the membrane lipids; instead of a slight alternating lipid arrangement in absence of Ca²⁺ due to the electrostatic interaction between the DPPS headgroups after addition of Ca²⁺ a lateral cluster structure is characteristic of the fluid phase.     

Intrinsic Variability in Shell and Soft Tissue Growth of the Freshwater Mussel Lampsilis siliquoidea

Freshwater mussels are ecologically and economically important members of many aquatic ecosystems, but are globally among the most imperiled taxa. Propagation techniques for mussels have been developed and used to boost declining and restore extirpated populations. Here we use a cohort of propagated mussels to estimate the intrinsic variability in size and growth rate of Lampsilis siliquoidea (a commonly propagated species). Understanding the magnitude and pattern of variation in data is critical to determining whether effects observed in nature or experimental treatments are likely to be important. The coefficient of variation (CV) of L. siliquoidea soft tissues (6.0%) was less than the CV of linear shell dimensions (25.1–66.9%). Size-weight relationships were best when mussel width (the maximum left-right dimension with both valves appressed) was used as a predictor, but 95% credible intervals on these predictions for soft tissues were ∼145 mg wide (about 50% of the mean soft tissue mass). Mussels in this study were treated identically, raised from a single cohort and yet variation in soft tissue mass at a particular size class (as determined by shell dimensions) was still high. High variability in mussel size is often acknowledged, but seldom discussed in the context of mussel conservation. High variability will influence the survival of stocked juvenile cohorts, may affect the ability to experimentally detect sublethal stressors and may lead to incongruities between the effects that mussels have on structure (via hard shells) and biogeochemical cycles (via soft tissue metabolism). Given their imperiled status and longevity, there is often reluctance to destructively sample unionid mussel soft tissues even in metabolic studies (e.g., studies of nutrient cycling). High intrinsic variability suggests that using shell dimensions (particularly shell length) as a response variable in studies of sublethal stressors or metabolic processes will make confident identifications of smaller effect sizes difficult.     

WIKI4, a Novel Inhibitor of Tankyrase and Wnt/?-Catenin Signaling

The Wnt/ß-catenin signaling pathway controls important cellular events during development and often contributes to disease when dysregulated. Using high throughput screening we have identified a new small molecule inhibitor of Wnt/ß-catenin signaling, WIKI4. WIKI4 inhibits expression of ß-catenin target genes and cellular responses to Wnt/ß-catenin signaling in cancer cell lines as well as in human embryonic stem cells. Furthermore, we demonstrate that WIKI4 mediates its effects on Wnt/ß-catenin signaling by inhibiting the enzymatic activity of TNKS2, a regulator of AXIN ubiquitylation and degradation. While TNKS has previously been shown to be the target of small molecule inhibitors of Wnt/ß-catenin signaling, WIKI4 is structurally distinct from previously identified TNKS inhibitors.     

A taxonomy of transcriptomic cell types across the isocortex and hippocampal formation

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Human Anti-Varicella-Zoster Virus (VZV) Recombinant Monoclonal Antibody Produced after Zostavax Immunization Recognizes the gH/gL Complex and Neutralizes VZV Infection

Varicella-zoster virus (VZV) is a ubiquitous, highly cell-associated, and exclusively human neurotropic alphaherpesvirus. VZV infection is initiated by membrane fusion, an event dependent in part on VZV glycoproteins gH and gL. Consistent with its location on the virus envelope, the gH/gL complex is a target of neutralizing antibodies produced after virus infection. One week after immunizing a 59-year-old VZV-seropositive man with Zostavax, we sorted his circulating blood plasma blasts and amplified expressed immunoglobulin variable domain sequences by single-cell PCR. Sequence analysis identified two plasma blast clones, one of which was used to construct a recombinant monoclonal antibody (rec-RC IgG). The rec-RC IgG colocalized with VZV gE on the membranes of VZV-infected cells and neutralized VZV infection in tissue culture. Mass spectrometric analysis of proteins immunoprecipitated by rec-RC IgG identified both VZV gH and gL. Transfection experiments showed that rec-RC IgG recognized a VZV gH/gL protein complex but not individual gH or gL proteins. Overall, our recombinant monoclonal anti-VZV antibody effectively neutralizes VZV and recognizes a conformational epitope within the VZV gH/L protein complex. An unlimited supply of this antibody provides the opportunity to analyze membrane fusion events that follow virus attachment and to identify multiple epitopes on VZV-specific proteins.