The ERG1A K+ Channel Is More Abundant in Rectus abdominis Muscle from Cancer Patients Than that from Healthy Humans.

BACKGROUND: The potassium channel encoded by the ether-a-gogo-related gene 1A (erg1a) has been detected in the atrophying skeletal muscle of mice experiencing either muscle disuse or cancer cachexia and further evidenced to contribute to muscle deterioration by enhancing ubiquitin proteolysis; however, to our knowledge, ERG1A has not been reported in human skeletal muscle. METHODS AND RESULTS: Here, using immunohistochemistry, we detect ERG1A immunofluorescence in human Rectus abdominis skeletal muscle sarcolemma. Further, using single point brightness data, we report the detection of ERG1A immunofluorescence at low levels in the Rectus abdominis muscle sarcolemma of young adult humans and show that it trends toward greater levels (10.6%) in healthy aged adults. Interestingly, we detect ERG1A immunofluorescence at a statistically greater level (53.6%; p < 0.05) in the skeletal muscle of older cancer patients than in age-matched healthy adults. Importantly, using immunoblot, we reveal that lower mass ERG1A protein is 61.5% (p < 0.05) more abundant in the skeletal muscle of cachectic older adults than in healthy age-matched controls. Additionally, we report that the ERG1A protein is detected in a cultured human rhabdomyosarcoma line that may be a good in vitro model for the study of ERG1A in muscle. CONCLUSIONS: The data demonstrate that ERG1A is detected more abundantly in the atrophied skeletal muscle of cancer patients, suggesting it may be related to muscle loss in humans as it has been shown to be in mice experiencing muscle atrophy as a result of malignant tumors.

Colorado tick fever virus induces apoptosis in human endothelial cells to facilitate viral replication.

Colorado tick fever virus (CTFV) belongs to the genus Coltivirus of the Reoviridae family, and it is the causative agent of Colorado tick fever. Symptoms of the infection are characterized by sudden biphasic fever, headache, and petechial rash, while severe forms of the disease can include meningoencephalitis, hemorrhagic fever, and death in children. However, the mechanisms underlying CTFV induced pathology and severe complications remain unknown. As CTFV is spread by tick bites and disseminates systemically via hematogenous routes, we performed in vitro analysis examining the interactions between endothelial cells (ECs) and CTFV. Our findings indicate that dermal microvascular ECs, HMEC-1, are susceptible and permissive to CTFV infection. To investigate the role of CTFV infection on endothelial barrier function, we assessed transendothelial electrical resistance (TEER) by xCELLigence and observed a dose-dependent decrease in cell index, indicating increased vascular permeability starting at approximately hour 18 (MOI=1) and hour 26 (MOI=0.1). Since CTFV induced cytopathic effect and increased vascular permeability in HMEC-1 cells, we hypothesized that CTFV causes apoptotic cell death. Our results showed that HMEC-1 cells infected with CTFV at 48 h caused a significant increase in Annexin V staining with reduced viability compared to uninfected cells suggesting CTFV induces apoptotic cell death in human ECs. Electron microscopy also was consistent with apoptotic features, including chromatin condensation and cell blebbing. Furthermore, CTFV induced caspase-3/7 activation at 24 and 48 h post-infection (hpi). The inhibition of caspase activity using Z-VAD-FMK reduced CTFV induced cell death and significantly reduced viral titer. These results indicated that CTFV can infect ECs, exerting direct adverse effects, leading to vascular permeability and cell death. Overall, our data suggest that caspase-mediated apoptosis is a critical mechanism by which CTFV induces disease in the host and enhances viral replication. Future studies will examine the viral and cellular determinants involved in CTFV induced apoptosis in human ECs.

Chemically Engineered Synthetic Lipid Vesicles for Sensing and Visualization of Protein-Bilayer Interactions.

From pathogen intrusion to immune response, the cell membrane plays an important role in signal transduction. Such signals are important for cellular proliferation and survival. However, measurement of these subtle signals through the lipid membrane scaffold is challenging. We present a chromatic model membrane vesicle system engineered to covalently bind with lysine residues of protein molecules for investigation of cellular interactions and signaling. We discovered that different protein molecules induced differential spectroscopic signals, which is based on the chemical and physical properties of protein interacting at the vesicle surface. The observed chromatic response (CR) for bound protein molecules with higher molecular weight was much larger (∼5-15×) than those for low molecular weight proteins. Through mass spectrometry (MS), we found that only 6 out of 60 (10%) lysine groups present in bovine serum albumin (BSA) were accessible to the membrane of the vesicles. Finally, a "sphere-shell" model representing the protein-vesicle complex was used for evaluating the contribution of van der Waals interactions between proteins and vesicles. Our analysis points to contributions from van der Waals, hydrophobic, and electrostatic interactions toward observed CR signals resulting from molecular interactions at the vesicle membrane surface. Overall, this study provided a convenient, chromatic, semiquantitative method of detecting biomolecules and their interactions with model membranes at sub-nanomolar concentration.

The sprightly little sphaerodactyl: Systematics and biogeography of the Puerto Rican dwarf geckos Sphaerodactylus (Gekkota, Sphaerodactylidae).

Studies of the Caribbean herpetofauna (amphibians and reptiles) have made significant contributions to our knowledge of evolutionary patterns and processes. A prerequisite for these studies are accurate taxonomies and robust phylogenetic hypotheses. One notable Caribbean radiation lacking such data are dwarf geckos of the genus Sphaerodactylus. Systematics of the Puerto Rican Sphaerodactylus have been turbulent since the initial species descriptions and no molecular phylogenies exist that include complete or near-complete taxon sampling. Here, we combine a multi-locus molecular phylogeny with extensive morphological information to investigate the current diversity of Sphaerodactylus geckos from the Puerto Rican Bank, with a large number of species from Hispaniola as an outgroup. In particular, we focus our efforts on resolving the taxonomy of the Sphaerodactylus macrolepis Günther species complex. We find S. macrolepis sensu lato (currently two nominal species with nine subspecies) is made up of at least four diagnosable species within two clades: (1) the sister species Sphaerodactylus macrolepis sensu stricto from the Virgin Islands (including St. Croix) and Culebra, and S. parvus King from islands in the northern Lesser Antilles; and (2) all other Sphaerodactylus macrolepis subspecies from Puerto Rico, Vieques, and Culebra. We resurrect Sphaerodactylus grandisquamis Stejneger from synonymy to refer to all subspecies from Puerto Rico and elevate the subspecies Sphaerodactylus inigoi Thomas Schwartz for geckos from Vieques and western Culebra. The resulting phylogeny and revised taxonomy will be a useful tool for subsequent research into Sphaerodactylus conservation and evolution.

Ground-State versus Excited-State Interchromophoric Interaction: Topology Dependent Excimer Contribution in Metal-Organic Framework Photophysics.

Metal-organic frameworks (MOFs) define emerging materials with unique optoelectronic properties that stem from the highly organized chromophoric linkers within their frameworks. The extent of ground- and excited-state interchromophoric interaction among the π-conjugated macrocyclic linkers was studied within three tetraphenyl-pyrene (1,3,6,8-tetrakis(p-benzoic acid)pyrene; H4TBAPy)-based MOFs: ROD-7 (In2(OH)2TBAPy, frz), NU-901 (scu), and NU-1000 (csq) via steady-state and time-resolved spectroscopic techniques. These experimental data along with computational results indicate that the extent of the interchromophoric interaction, leading to a reduced optical band gap, varies across the series of MOFs and is a function of the relative orientation of the TBAPy linkers determined by their respective framework topology. The trend in the S1 → S0 emission lifetime is consistent with their relative optical bandgap. Analyses of the transient emission decay profiles and time-resolved emission spectroscopic data, recorded in low dielectric media, reveal that a long-lived emissive excimer state appears ∼1850 ± 150 cm-1 lower in energy relative to their corresponding S1 → S0 transitions. The emissive contribution from this excimer state, as well as its corresponding transition energy and time constants, are also found to be dependent on MOF identity. Such variation in properties are particularly influenced by the number density of the TBAPy linkers presented by the topology of a given MOF that are primed to form such an excited state complex. The present work shows how the specific arrangement of the linkers can play a key role in the photophysical properties of MOFs.

Topology-dependent emissive properties of zirconium-based porphyrin MOFs.

Highly ordered chromophoric linkers positioned within the metal-organic frameworks (MOFs) have the potential to mimic natural light-harvesting complexes. Herein we report topological control over the photophysical properties of MOFs via modular interchromophoric electronic coupling to manifest different steady-state singlet emission spectra and their corresponding fluorescence lifetimes.