Demonstration of the protective effects of fluorescent proteins in baculoviruses exposed to ultraviolet light inactivation

Autographa californica multiple nucleopolyhedrovirus (AcMNPV) recombinants, namely AcRFP produced by fusion of the red fluorescent protein (RFP) gene with the polyhedrin gene, and a recombinant (pAcUW21-23GFP) carrying the green fluorescent protein (GFP) in its viral envelope, were evaluated for their resistance to inactivation by ultraviolet light. AcRFP recombinants produced incomplete polyhedra with low infectivity for Trichoplusia ni larvae, whereas AcuW21-23GFP produced normal polyhedra with high infectivity. Electron microscopy of AcRFP CL14 showed the incorporation of very few viral particles into polyhedrin matrix protein material. The LC(50) for AcuW21-23GFP was 0.10 occlusion bodies/mm(2), whereas the LC(50) values for several AcRFP recombinants ranged from 20 to 329 occlusion bodies/mm(2). When both the RFP and GFP recombinants were exposed to ultraviolet light (UV-B 280–320 nm), the results support the conclusion that these fluorescent proteins afford some protection against its damaging effects. Abbreviation: / AcMNPV: Autographa californica multiple nucleopolyhedrovirus BV: budded virus CPE: cytopathogenic effect ECV: extracellular virus OB: occlusion body ODV: occlusion derived virus RFP: red fluorescent protein GFP: green fluorescent protein TCID(50): tissue culture infective dose at the 50 % level UV-B: ultraviolet light of 280–320 n

Identification of BV/ODV-C42, an Autographa californica Nucleopolyhedrovirus orf101-Encoded Structural Protein Detected in Infected-Cell Complexes with ODV-EC27 and p78/83

orf101 is a late gene of Autographa californica nucleopolyhedrovirus (AcMNPV). It encodes a protein of 42 kDa which is a component of the nucleocapsid of budded virus (BV) and occlusion-derived virus (ODV). To reflect this viral localization, the product of orf101 was named BV/ODV-C42 (C42). C42 is predominantly detected within the infected-cell nucleus: at 24 h postinfection (p.i.), it is coincident with the virogenic stroma, but by 72 h p.i., the stroma is minimally labeled while C42 is more uniformly located throughout the nucleus. Yeast two-hybrid screens indicate that C42 is capable of directly interacting with the viral proteins p78/83 (1629K) and ODV-EC27 (orf144). These interactions were confirmed using blue native gels and Western blot analyses. At 28 h p.i., C42 and p78/83 are detected in two complexes: one at approximately 180 kDa and a high-molecular-mass complex (500 to 600 kDa) which also contains EC27

Structural Relationships Across the Sevier Gravity Slide of Southwest Utah and Implications for Catastrophic Translation and Emplacement Processes of Long Runout Landslides

The physical processes that facilitate long-distance translation of large-volume gravity slides remain poorly understood. To better understand these processes and the controls on runout distance, we conducted an outcrop and microstructural characterization of the Sevier gravity slide across the former land surface and summarize findings of four key sites. The Sevier gravity slide is the oldest of three mega-scale (\u3e1,000 km2) collapse events of the Marysvale volcanic field (Utah, USA). Field observations of intense deformation, clastic dikes, pseudotachylyte, and consistency of kinematic indicators support the interpretation of rapid emplacement during a single event. Furthermore, clastic dikes and characteristics of the slip zone suggest emplacement involved mobilization and pressurized injection of basal material. Across the runout distance, we observe evidence for progressive slip delocalization along the slide base. This manifests as centimeter- to decimeter-thick cataclastic basal zones and abundant clastic dikes in the north and tens of meters thick basal zones characterized by widespread deformation of both slide blocks and underlying rock near the southern distal end of the gravity slide. Superimposed on this transition are variations in basal zone characteristics and slide geometry arising from interactions between slide blocks during dynamic wear and deposition processes and pre-existing topography of the former land surface. These observations are synthesized into a conceptual model in which the presence of highly pressurized fluids reduced the frictional resistance to sliding during the emplacement of the Sevier gravity slide, and basal zone evolution controlled the effectiveness of dynamic weakening mechanisms across the former land surface

The Concept of Tectonic Provenance: Case Study of the Gigantic Markagunt Gravity Slide Basal Layer

Formation and evolution of the basal layer in large landslides has important implications for processes that reduce frictional resistance to sliding. In this report, we show that zircon geochronology and tectonic provenance can be used to investigate the basal layer of the gigantic-scale Markagunt gravity slide of Utah, USA. Basal layer and clastic injectite samples have unique tectonic chronofacies that identify the rock units that were broken down during emplacement. Our results show that basal material from sites on the former land surface is statistically indistinguishable and formed primarily by the breakdown of upper plate lithologies during sliding. Decapitated injectites have a different tectonic chronofacies than the local basal layer, with more abundant lower plate-derived zircons. This suggests clastic dikes formed earlier in the translation history from a structurally deeper portion of the slide surface and a compositionally different basal layer before being translated to their current position

X-Ray Phase-Contrast Tomography of Renal Ischemia-Reperfusion Damage

Purpose: The aim of the study was to investigate microstructural changes occurring in unilateral renal ischemia-reperfusion injury in a murine animal model using synchrotron radiation. Material and Methods: The effects of renal ischemia-reperfusion were investigated in a murine animal model of unilateral ischemia. Kidney samples were harvested on day 18. Grating-Based Phase-Contrast Imaging (GB-PCI) of the paraffin-embedded kidney samples was performed at a Synchrotron Radiation Facility (beam energy of 19 keV). To obtain phase information, a two-grating Talbot interferometer was used applying the phase stepping technique. The imaging system provided an effective pixel size of 7.5 mu m. The resulting attenuation and differential phase projections were tomographically reconstructed using filtered back-projection. Semi-automated segmentation and volumetry and correlation to histopathology were performed. Results: GB-PCI provided good discrimination of the cortex, outer and inner medulla in non-ischemic control kidneys. Post-ischemic kidneys showed a reduced compartmental differentiation, particularly of the outer stripe of the outer medulla, which could not be differentiated from the inner stripe. Compared to the contralateral kidney, after ischemia a volume loss was detected, while the inner medulla mainly retained its volume (ratio 0.94). Post-ischemic kidneys exhibited severe tissue damage as evidenced by tubular atrophy and dilatation, moderate inflammatory infiltration, loss of brush borders and tubular protein cylinders. Conclusion: In conclusion GB-PCI with synchrotron radiation allows for non-destructive microstructural assessment of parenchymal kidney disease and vessel architecture. If translation to lab-based approaches generates sufficient density resolution, and with a time-optimized image analysis protocol, GB-PCI may ultimately serve as a non-invasive, non-enhanced alternative for imaging of pathological changes of the kidney

Nail lacquer films’ surface energies and in vitro water-resistance and adhesion do not predict their in vivo residence

The in vivo residence of nail lacquers (which are ideal topical drug carriers for the treatment of nail diseases) determines their frequency of application, and is thereby expected to influence patient adherence and success of treatment. Thus in vitro measurements to indicate lacquers’ in vivo residence are routinely conducted during formulation development. However the literature on in vitro-in vivo correlations is severely limited. Thus, the aim of the work discussed in this paper was to investigate correlations between in vivo residence and in vitro film resistance to water, in vitro film adhesion and surface energy of lacquer films. In vivo measurements were conducted on fingernails in six volunteers. Seven commercially available nail lacquers were tested in commonly-used measurements. Correlations between in vivo residence and in vitro water resistance and adhesion were found to be extremely poor. The surface energies of the lacquer films (which were between 33 and 39 mJ/m2) were also not predictive of in vivo residence. High density polyethylene (HDPE) sheet – whose surface energy was determined to be similar to that of the human nailplate – was found to be a suitable model for the nailplate (when investigating surface energy) and was used in a number of experiments

Genomic Sequence Analysis of Granulovirus Isolated from the Tobacco Cutworm, Spodoptera litura

Background: Spodoptera litura is a noctuid moth that is considered an agricultural pest. The larvae feed on a wide range of plants and have been recorded on plants from 40 plant families (mostly dicotyledons). It is a major pest of many crops. To better understand Spodoptera litura granulovirus (SpliGV), the nucleotide sequence of the SpliGV DNA genome was determined and analyzed. Methodology/Principal Findings: The genome of the SpliGV was completely sequenced. The nucleotide sequence of the SpliGV genome was 124,121 bp long with 61.2 % A+T content and contained 133 putative open reading frames (ORFs) of 150 or more nucleotides. The 133 putative ORFs covered 86.3 % of the genome. Among these, 31 ORFs were conserved in most completely sequenced baculovirus genomes, 38 were granulovirus (GV)-specific, and 64 were present in some nucleopolyhedroviruses (NPVs) and/or GVs. We proved that 9 of the ORFs were SpliGV specific. Conclusions/Significance: The genome of SpliGV is 124,121 bp in size. One hundred thirty-three ORFs that putatively encode proteins of 50 or more amino acid residues with minimal overlap were determined. No chitinase or cathepsin genes, which are involved in the liquefaction of the infected host, were found in the SpliGV genome, explaining why SpliGVinfected insects do not degrade in a typical manner. The DNA photolyase gene was first found in the genus Granulovirus. When phylogenic relationships were analyzed, the SpliGV was most closely related to Trichoplusia ni granulovirus (TnGV

The Salivary Secretome of the Tsetse Fly Glossina pallidipes (Diptera: Glossinidae) Infected by Salivary Gland Hypertrophy Virus

Tsetse fly (Diptera; Glossinidae) transmits two devastating diseases to farmers (human African Trypanosomiasis; HAT) and their livestock (Animal African Trypanosomiasis; AAT) in 37 sub-Saharan African countries. During the rainy seasons, vast areas of fertile, arable land remain uncultivated as farmers flee their homes due to the presence of tsetse. Available drugs against trypanosomiasis are ineffective and difficult to administer. Control of the tsetse vector by Sterile Insect Technique (SIT) has been effective. This method involves repeated release of sterilized males into wild tsetse populations, which compete with wild type males for females. Upon mating, there is no offspring, leading to reduction in tsetse populations and thus relief from trypanosomiasis. The SIT method requires large-scale tsetse rearing to produce sterile males. However, tsetse colony productivity is hampered by infections with the salivary gland hypertrophy virus, which is transmitted via saliva as flies take blood meals during membrane feeding and often leads to colony collapse. Here, we investigated the salivary gland secretome proteins of virus-infected tsetse to broaden our understanding of virus infection, transmission and pathology. By this approach, we obtain insight in tsetse-hytrosavirus interactions and identified potential candidate proteins as targets for developing biotechnological strategies to control viral infections in tsetse colonies