Resistance of the 64K protein of budded Autographa californica nuclear polyhedrosis virus to functional inactivation by proteolysis.

The 64K surface protein of budded Autographa californica nuclear polyhedrosis virus (AcMNPV BV) is known to play a role in the functional entry of AcMNPV BV into Spodoptera frugiperda IPLB-SF-21 cells by adsorptive endocytosis. AcV1, a neutralizing monoclonal antibody, reacts with the 64K protein and in doing so prevents efficient entry. In this communication we report that treatment of AcMNPV BV with either trypsin or proteinase K cleaves the 64K protein into one major fragment of 34.6K and two minor fragments of 36K to 37.2K that are retained with the virus. All of the fragments are glycosylated. Protease treatment does not reduce viral infectivity, but it does result in the destruction of the AcV1-reactive epitope; thus AcV1 is not able to neutralize protease-treated AcMNPV BV. Polyclonal antiserum to BV is able to recognize both cleaved and uncleaved 64K and neutralize both protease-treated and untreated virus. Protease treatment does not diminish the sensitivity of AcMNPV BV to chloroquine, but it does cause the virus to become more susceptible to inactivation by 2-mercaptoethanol (2-ME) even though exposure to 2-ME does not result in dissociation of the fragments from the virus

Budded Autographa californica NPV 64K protein: Further biochemical analysis and effects of postimmunoprecipitation sample preparation conditions.

Previously it was shown that AcV1, a neutralizing monoclonal antibody of the Autographa californica nuclear polyhedrosis virus-budded phenotype reacted with a surface antigen present on infected cells during virus budding, and in the viral envelope (L. E. Volkman, P. A. Goldsmith, R. T. Hess, and P. Faulkner (1984), Virology 133, 354-362). Radioimmune precipitation of solubilized, [35S]methionine-labeled budded virus with AcV1 and analysis on SDS-PAGE revealed four bands consistently: one major band at 64,000 Da, and three minor bands at 127,000, 59,000, and 49,000 Da. The reason for the appearance of four bands instead of one was unclear. Data suggest that two of the bands, 49K and 59K, are aberrant, and are the products of sample preparation conditions. Further, evidence is presented that the 127K band is composed of dimers of the 64K protein, and that under nonreducing conditions, oligomers (trimers and tetramers) of 64K protein can also be detected. BVGP 64 is additionally shown to be phosphorylated and to have an isoelectric point of 3.15. The BVGP 64 epitope reactive with AcV1 is destroyed by interaction with SDS. This could account for the lack of neutralizing activity of antiserum made to the SDS-PAGE purified BVGP 64

Neutralization of budded Autographa californica NPV by a monoclonal antibody: identification of the target antigen.

A neutralizing monoclonal antibody of the budded phenotype of Autographa californica nuclear polyhedrons virus did not react with the occluded form of the virus as determined by neutralization, ELISA, and indirect immunoperoxidase staining. The antibody did react with the surface of infected cells in the prepolyhedra stage of cytopathic effect, the period of active virus budding. Immunoelectron microscopy indicated the antigen(s) reactive with the neutralizing antibody was present all around the viral envelope, but was more highly concentrated at the end with peplomers. Four proteins were immunoprecipitated from solubilized, radiolabeled budded virus preparations with the monoclonal antibody. The major protein had a molecular weight of approximately 64,000, while the other three were approximately 127,000, 59,000, and 49,000. All four proteins could be labeled with N-acetyl-d-[1-3H]glucosamine. This glycosylation reaction could be inhibited by tunicamycin

Container size exerts a stronger influence than liquid volume on the perceived weight of objects

This is the author accepted manuscript. The final version is available from Elsevier via the DOI in this recordMany features of an object can influence how we predict and perceive its weight. The current study evaluated the relative contributions of sensory and conceptual processing of object features on weight perception. We employed a novel paradigm to investigate how container size and the amount of liquid inside can influence the perceived weight of bottles and the forces deployed when lifting them. Stimulus pairs always had the same mass but could vary in liquid volume (full vs half-full bottle) or size (large vs small bottle; size-weight illusion (SWI)). In Experiment 1, participants lifted the stimuli via strings, which served to isolate the influence of visual from kinaesthetic information about the size of stimuli on perception and lifting behaviour. In Experiment 2, participants lifted the stimuli via handles that were attached directly to the objects. This lifting style is more likely to include deviations from true vertical lifting, which should theoretically provide more kinaesthetic information about the size of the stimuli. Experiment 1 did not produce any weight illusion. Experiment 2 produced a weight illusion but only when container size differed. Thus, liquid volume did not influence perceived weight when container size was held constant in either experiment. Curiously, additional control experiments revealed that participants could not discriminate between the different sized bottles solely from the kinaesthetic information received from a handle-based lift, suggesting that size might be processed differently when making explicit perceptual judgements about it than when influencing weight perception. Together, these findings suggest that weight illusions are driven more strongly by the kinaesthetic processing of stimulus features than predictions arising from conceptual weight cues.Australian Research Council (ARC

Maximal Spontaneous Photon Emission and Energy Loss from Free Electrons

Free electron radiation such as Cerenkov, Smith--Purcell, and transition radiation can be greatly affected by structured optical environments, as has been demonstrated in a variety of polaritonic, photonic-crystal, and metamaterial systems. However, the amount of radiation that can ultimately be extracted from free electrons near an arbitrary material structure has remained elusive. Here we derive a fundamental upper limit to the spontaneous photon emission and energy loss of free electrons, regardless of geometry, which illuminates the effects of material properties and electron velocities. We obtain experimental evidence for our theory with quantitative measurements of Smith--Purcell radiation. Our framework allows us to make two predictions. One is a new regime of radiation operation---at subwavelength separations, slower (nonrelativistic) electrons can achieve stronger radiation than fast (relativistic) electrons. The second is a divergence of the emission probability in the limit of lossless materials. We further reveal that such divergences can be approached by coupling free electrons to photonic bound states in the continuum (BICs). Our findings suggest that compact and efficient free-electron radiation sources from microwaves to the soft X-ray regime may be achievable without requiring ultrahigh accelerating voltages.Comment: 7 pages, 4 figure

Effect of tube diameter and capillary number on platelet margination and near-wall dynamics

The effect of tube diameter $D$ and capillary number $Ca$ on platelet margination in blood flow at $\approx 37\%$ tube haematocrit is investigated. The system is modelled as three-dimensional suspension of deformable red blood cells and nearly rigid platelets using a combination of the lattice-Boltzmann, immersed boundary and finite element methods. Results show that margination is facilitated by a non-diffusive radial platelet transport. This effect is important near the edge of the cell-free layer, but it is only observed for $Ca > 0.2$, when red blood cells are tank-treading rather than tumbling. It is also shown that platelet trapping in the cell-free layer is reversible for $Ca \leq 0.2$. Only for the smallest investigated tube ($D = 10 \mu\text{m}$) margination is essentially independent of $Ca$. Once platelets have reached the cell-free layer, they tend to slide rather than tumble. The tumbling rate is essentially independent of $Ca$ but increases with $D$. Tumbling is suppressed by the strong confinement due to the relatively small cell-free layer thickness at $\approx 37\%$ tube haematocrit.Comment: 16 pages, 10 figure

Rates and risks for prolonged grief disorder in a sample of orphaned and widowed genocide survivors

<p>Abstract</p> <p>Background</p> <p>The concept of Prolonged Grief Disorder (PGD) has been defined in recent years by Prigerson and co-workers, who have developed and empirically tested consensus and diagnostic criteria for PGD. Using these most recent criteria defining PGD, the aim of this study was to determine rates of and risks for PGD in survivors of the 1994 Rwandan genocide who had lost a parent and/or the husband before, during or after the 1994 events.</p> <p>Methods</p> <p>The PG-13 was administered to 206 orphans or half orphans and to 194 widows. A regression analysis was carried out to examine risk factors of PGD.</p> <p>Results</p> <p>8.0% (<it>n </it>= 32) of the sample met criteria for PGD with an average of 12 years post-loss. All but one person had faced multiple losses and the majority indicated that their grief-related loss was due to violent death (70%). Grief was predicted mainly by time since the loss, by the violent nature of the loss, the severity of symptoms of posttraumatic stress disorder (PTSD) and the importance given to religious/spiritual beliefs. By contrast, gender, age at the time of bereavement, bereavement status (widow versus orphan), the number of different types of losses reported and participation in the funeral ceremony did not impact the severity of prolonged grief reactions.</p> <p>Conclusions</p> <p>A significant portion of the interviewed sample continues to experience grief over interpersonal losses and unresolved grief may endure over time if not addressed by clinical intervention. Severity of grief reactions may be associated with a set of distinct risk factors. Subjects who lose someone through violent death seem to be at special risk as they have to deal with the loss experience as such and the traumatic aspects of the loss. Symptoms of PTSD may hinder the completion of the mourning process. Religious beliefs may facilitate the mourning process and help to find meaning in the loss. These aspects need to be considered in the treatment of PGD.</p

The HDAC Inhibitor FK228 Enhances Adenoviral Transgene Expression by a Transduction-Independent Mechanism but Does Not Increase Adenovirus Replication

The histone deacetylase inhibitor FK228 has previously been shown to enhance adenoviral transgene expression when cells are pre-incubated with the drug. Upregulation of the coxsackie adenovirus receptor (CAR), leading to increased viral transduction, has been proposed as the main mechanism. In the present study, we found that the highest increase in transgene expression was achieved when non-toxic concentrations of FK228 were added immediately after transduction, demonstrating that the main effect by which FK228 enhances transgene expression is transduction-independent. FK228 had positive effects both on Ad5 and Ad5/f35 vectors with a variety of transgenes and promoters, indicating that FK228 works mainly by increasing transgene expression at the transcriptional level. In some cases, the effects were dramatic, as demonstrated by an increase in CD40L expression by FK228 from 0.3% to 62% when the murine prostate cancer cell line TRAMP-C2 was transduced with Ad[CD40L]. One unexpected finding was that FK228 decreased the transgene expression of an adenoviral vector with the prostate cell-specific PPT promoter in the human prostate adenocarcinoma cell lines LNCaP and PC-346C. This is probably a consequence of alteration of the adenocarcinoma cell lines towards a neuroendocrine differentiation after FK228 treatment. The observations in this study indicate that FK228 enhances adenoviral therapy by a transduction-independent mechanism. Furthermore, since histone deacetylase inhibitors may affect the differentiation of cells, it is important to keep in mind that the activity and specificity of tissue- and tumor-specific promoters may also be affected

Predicting Weight Outcomes in Preadolescence: The Role of Toddlers? Self-regulation Skills and the Temperament Dimension of Pleasure

Objective To investigate the role of toddlers? self-regulation skills and temperament in predicting weight outcomes in preadolescence. Method Participants for this study included 195 children (114 girls) obtained from three different cohorts participating in a larger ongoing longitudinal study. At 2 years of age, participants participated in several laboratory tasks designed to assess their self-regulation abilities, including emotion regulation, sustained attention, and delay of gratification, while parents filled out a temperament questionnaire to assess toddlers? pleasure expression. Height and weight measures were collected when children were 4, 5, 7, and 10 years of age. Children also filled out a body image and eating questionnaire at the 10 year visit. Results Self-regulation skills in toddlers were associated with both BMI development, pediatric obesity, and body image/eating concerns. The temperament dimension of pleasure was also associated with BMI development and pediatric obesity but not body image/eating concerns. Conclusion Self-regulation difficulties across domains as well as temperament based pleasure in toddlers represented significant individual risk factors for the development of pediatric obesity eight years later. Early self-regulation difficulties also contributed to body image and eating concerns that typically accompanied overweight children. The mechanisms by which early self-regulation skills and temperament based pleasure may contribute to the development of pediatric obesity and associated weight concerns are discussed