The recent emergence of highly virulent human adenoviruses (HAdVs) with new tissue tropisms underscores the need to determine their ontogeny. Here we report complete high quality genome sequences and analyses for all the previously unsequenced HAdV serotypes (n = 20) within HAdV species D. Analysis of nucleotide sequence variability for these in conjunction with another 40 HAdV prototypes, comprising all seven HAdV species, confirmed the uniquely hypervariable regions within species. The mutation rate among HAdV-Ds was low when compared to other HAdV species. Homologous recombination was identified in at least two of five examined hypervariable regions for every virus, suggesting the evolution of HAdV-Ds has been highly dependent on homologous recombination. Patterns of alternating GC and AT rich motifs correlated well with hypervariable region recombination sites across the HAdV-D genomes, suggesting foci of DNA instability lead to formulaic patterns of homologous recombination and confer agility to adenovirus evolution.

Hydrogenosomes and mitosomes are mitochondrion-related organelles in anaerobic/microaerophilic eukaryotes with highly reduced and divergent functions. The full diversity of their content and function, however, has not been fully determined. To understand the central role of mitosomes in Entamoeba histolytica, a parasitic protozoon that causes amoebic dysentery and liver abscesses, we examined the proteomic profile of purified mitosomes. Using 2 discontinuous Percoll gradient centrifugation and MS analysis, we identified 95 putative mitosomal proteins. Immunofluorescence assay showed that 3 proteins involved in sulfate activation, ATP sulfurylase, APS kinase, and inorganic pyrophosphatase, as well as sodium/sulfate symporter, involved in sulfate uptake, were compartmentalized to mitosomes. We have also provided biochemical evidence that activated sulfate derivatives, adenosine-5-phosphosulfate and 3-phosphoadenosine-5-phosphosulfate, were produced in mitosomes. Phylogenetic analysis showed that the aforementioned proteins and chaperones have distinct origins, suggesting the mosaic character of mitosomes in E. histolytica consisting of proteins derived from ␣-proteobacterial, ␦-proteobacterial, and ancestral eukaryotic origins. These results suggest that sulfate activation is the major function of mitosomes in E. histolytica and that E. histolytica mitosomes represent a unique mitochondrion-related organelle with remarkable diversity.anaerobic protozoa ͉ evolution ͉ mitochondria ͉ organelle ͉ proteomics D iversification of mitochondrial structure and function has occurred during eukaryotic evolution, and was especially observed in anaerobic/microaerophilic environments. Most extant anaerobic eukaryotes, which were previously considered to lack mitochondria, are now regarded to possess reduced and highly divergent forms of mitochondrion-related organelles (1, 2). The hydrogenosome is an organelle in which hydrogen and ATP are produced, and is found in anaerobic protists and fungi such as Trichomonas vaginalis (3, 4), Neocallimastix patriciarum (5, 6), and Nyctotherus ovalis (7). The mitosome, typically demonstrated in parasitic and free-living protists such as E. histolytica (2,(8)(9)(10)(11)(12)(13)(14)(15), Giardia intestinalis (16, 17), diverse microsporidian species (18)(19)(20), and Cryptosporidium parvum (21), generally has reduced functions and does not produce hydrogen or ATP. In Mastigamoeba balamuthi, a mitochondrion-related organelle was discovered and presumed to possess a unique array of biochemical properties, although it remains unclear whether the organelle is more similar to either hydrogenosomes or mitosomes (22). In contrast, the mitochondrion-related organelle in Blastocystis contains DNA and shows characteristics for both hydrogenosomes and mitochondria of higher eukaryotes (23). Organisms that possess hydrogenosomes and mitosomes do not cluster together in eukaryote phylogenies, indicating that secondary losses and changes in mitochondrial functions have independently occurred multiple times i...

A series of binary (HgSe) and ternary (HgSe1 - x S x ) mercury chalcogenide clusters are synthesized utilizing a colloidal technique involving the phase separation of metal and chalcogen precursors in the presence of strong Hg(II) coordinating ligands. The clusters vary in size between 2 and 3 nm and possess the cubic zinc blende structure of the bulk. Energy-dispersive X-ray measurements show that the composition of the ternary material can be varied throughout the entire composition range from HgS to HgSe. In all cases, the linear absorption of these binary and ternary species is narrow with well-resolved transitions at both the band edge and at higher energies. Complementary band edge emission is also observed with no apparent deep trap emission. Size- and composition (x)-dependent optical properties of these clusters are investigated using photoluminescence (PL) and photoluminescence excitation (PLE) spectra. In the case of HgSe clusters, the size-dependent behavior of up to four excited states is followed. For HgSe1 - x S x clusters, where x varies from 0 to 1, a size/composition-dependent progression of up to five excited states is observed.

The cellular entry of viruses represents a critical area of study, not only for viral tropism, but also because viral entry dictates the nature of the immune response elicited upon infection. Epidemic keratoconjunctivitis (EKC), caused by viruses within human adenovirus species D (HAdV-D), is a severe, ocular surface infection associated with corneal inflammation. Clathrin-mediated endocytosis has previously been shown to play a critical role in entry of other HAdV species into many host cell types. However, HAdV-D endocytosis into corneal cells has not been extensively studied. Herein, we show an essential role for cholesterol rich, lipid raft microdomains and caveolin-1, in the entry of HAdV-D37 into primary human corneal fibroblasts. Cholesterol depletion using methyl-β-cyclodextrin (MβCD) profoundly reduced viral infection. When replenished with soluble cholesterol, the effect of MβCD was reversed, allowing productive viral infection. HAdV-D37 DNA was identified in caveolin-1 rich endosomal fractions after infection. Src kinase activity was also increased in caveolin-1 rich endosomal fractions after infection, and Src phosphorylation and CXCL1 induction were both decreased in caveolin-1-/- mice corneas compared to wild type mice. siRNA knock down of caveolin-1 in corneal cells reduced chemokine induction upon viral infection, and caveolin-1-/- mouse corneas showed reduced cellular entry of HAdV-D37. As a control, HAdV-C2, a non-corneal pathogen, appeared to utilize the caveolar pathway for entry into A549 cells, but failed to infect corneal cells entirely, indicating virus and cell specific tropism. Immuno-electron microscopy confirmed the presence of caveolin-1 in HAdV-D37-containing vesicles during the earliest stages of viral entry. Collectively, these experiments indicate for the first time that HAdV-D37 uses a lipid raft mediated caveolin-1 associated pathway for entry into corneal cells, and connects the processes of viral entry with downstream proinflammatory cell signaling.

High-quality colloidal mercury sulfide quantum dots (QDs) are synthesized at room temperature using a strategy combining the effects of strongly binding Hg(II) ligands and metal/chalcogen precursor phase separation. This combination prevents both the rapid precipitation of bulk HgS in preparations involving only weak Hg(II) ligands and the reduction of mercury that takes place when only strongly binding ligands are used to slow the growth kinetics. Both the linear absorption and complementary band edge emission of the synthesized HgS QDs exhibit narrow, size-dependent transitions between 500 and 800 nm for sizes ranging from 1 to 5 nm in diameter. The metastable zinc blende phase of HgS is verified by wide-angle X-ray diffraction experiments and suggests potentially large tunable band edges if larger HgS nanocrystals that approach the bulk (zero energy) gap can be made. Growth of HgS QDs can be arrested by subsequent addition of Cd or Zn to the surface, after which the QDs can be stabilized with long-chain thiols or amines.

Direct or indirect exposure to an explosion can induce traumatic brain injury (TBI) of various severity levels. Primary TBI from blast exposure is commonly characterized by internal injuries, such as vascular damage, neuronal injury, and contusion, without external injuries. Current animal models of blast-induced TBI (bTBI) have helped to understand the deleterious effects of moderate to severe blast forces. However, the neurological effects of mild blast forces remain poorly characterized. Here, we investigated the effects caused by mild blast forces combining neuropathological, histological, biochemical and neurophysiological analysis. For this purpose, we employed a rodent blast TBI model with blast forces below the level that causes macroscopic neuropathological changes. We found that mild blast forces induced neuroinflammation in cerebral cortex, striatum and hippocampus. Moreover, mild blast triggered microvascular damage and axonal injury. Furthermore, mild blast caused deficits in hippocampal short-term plasticity and synaptic excitability, but no impairments in long-term potentiation. Finally, mild blast exposure induced proteolytic cleavage of spectrin and the cyclin-dependent kinase 5 activator, p35 in hippocampus. Together, these findings show that mild blast forces can cause aberrant neurological changes that critically impact neuronal functions. These results are consistent with the idea that mild blast forces may induce subclinical pathophysiological changes that may contribute to neurological and psychiatric disorders.

For DNA viruses, genetic recombination, addition, and deletion represent important evolutionary mechanisms. Since these genetic alterations can lead to new, possibly severe pathogens, we applied a systems biology approach to study the pathogenicity of a novel human adenovirus with a naturally occurring deletion of the canonical penton base Arg-Gly-Asp (RGD) loop, thought to be critical to cellular entry by adenoviruses. Bioinformatic analysis revealed a new highly recombinant species D human adenovirus (HAdV-D60). A synthesis of in silico and laboratory approaches revealed a potential ocular tropism for the new virus. In vivo, inflammation induced by the virus was dramatically greater than that by adenovirus type 37, a major eye pathogen, possibly due to a novel alternate ligand, Tyr-Gly-Asp (YGD), on the penton base protein. The combination of bioinformatics and laboratory simulation may have important applications in the prediction of tissue tropism for newly discovered and emerging viruses.

Variable temperature x-ray diffraction and differential scanning calorimetry experiments on the superlattice solids of alkanethiol protected silver nanoclusters show that the translational periodicity collapses around 398 K resulting in a liquid phase, which upon cooling, reverts into the parent crystalline phase. This reversibility is seen in a narrow temperature window of 400-448 K. If the heating is done above 473 K, the reversibility is lost, and the liquid in the cooling cycle freezes into a disordered phase. The stability of different phases, encountered in the experiments, is discussed in terms of a phenomenological free energy.