CNS activity of Pokeweed Anti-viral Protein (PAP) in mice infected with Lymphocytic Choriomeningitis Virus (LCMV)

BACKGROUND: Others and we have previously described the potent in vivo and in vitro activity of the broad-spectrum antiviral agent PAP (Pokeweed antiviral protein) against a wide range of viruses. The purpose of the present study was to further elucidate the anti-viral spectrum of PAP by examining its effects on the survival of mice challenged with lymphocytic choriomeningitis virus (LCMV). METHODS: We examined the therapeutic effect of PAP in CBA mice inoculated with intracerebral injections of the WE54 strain of LCMV at a 1000 PFU dose level that is lethal to 100% of mice within 7–9 days. Mice were treated either with vehicle or PAP administered intraperitoneally 24 hours prior to, 1 hour prior to and 24 hours, 48 hours 72 hours and 96 hours after virus inoculation. RESULTS: PAP exhibits significant in vivo anti- LCMV activity in mice challenged intracerebrally with an otherwise invariably fatal dose of LCMV. At non-toxic dose levels, PAP significantly prolonged survival in the absence of the majority of disease-associated symptoms. The median survival time of PAP-treated mice was >21 days as opposed to 7 days median survival for the control (p = 0.0069). CONCLUSION: Our results presented herein provide unprecedented experimental evidence that PAP exhibits antiviral activity in the CNS of LCMV-infected mice

Photometric variability in the old open cluster M 67. II. General Survey

We use differential CCD photometry to search for variability in BVI among 990 stars projected in and around the old open cluster M 67. In a previous paper we reported results for 22 cluster members that are optical counterparts to X-ray sources; this study focuses on the other stars in our observations. A variety of sampling rates were employed, allowing variability on time scales ranging from \sim 0.3 hours to \sim 20 days to be studied. Among the brightest sources studied, detection of variability as small as sigma approx 10 mmag is achieved (with > 3 sigma confidence); for the typical star observed, sensitivity to variability at levels sigma approx 20 mmag is achieved. The study is unbiased for stars with 12.5 < B < 18.5, 12.5 < V < 18.5, and 12 < I < 18 within a radius of about 10 arcmin from the cluster centre. In addition, stars with 10 < BVI < 12.5 were monitored in a few small regions in the cluster. We present photometry for all 990 sources studied, and report the variability characteristics of those stars found to be variable at a statistically significant level. Among the variables, we highlight several sources that merit future study, including stars located on the cluster binary sequence, stars on the giant branch, blue stragglers, and a newly discovered W UMa system.Comment: 12 pages, including 6 figures and 5 tables. Tables 1 and 3 only available in electronic version of paper. Accepted by A&

Sub-Subgiants in the Old Open Cluster M67?

We report the discovery of two spectroscopic binaries in the field of the old open cluster M67 -- S1063 and S1113 -- whose positions in the color-magnitude diagram place them approximately 1 mag below the subgiant branch. A ROSAT study of M67 independently discovered these stars to be X-ray sources. Both have proper-motion membership probabilities greater than 97%; precise center-of-mass velocities are consistent with the cluster mean radial velocity. S1063 is also projected within one core radius of the cluster center. S1063 is a single-lined binary with a period of 18.396 days and an orbital eccentricity of 0.206. S1113 is a double-lined system with a circular orbit having a period of 2.823094 days. The primary stars of both binaries are subgiants. The secondary of S1113 is likely a 0.9 Mo main-sequence star, which implies a 1.3 Mo primary star. We have been unable to explain securely the low apparent luminosities of the primary stars; neither binary contain stars presently limited in radius by their Roche lobes. We speculate that S1063 and S1113 may be the products of close stellar encounters involving binaries in the cluster environment, and may define alternative stellar evolutionary tracks associated with mass-transfer episodes, mergers, and/or dynamical stellar exchanges

In vitro template-change PCR to create single crossover libraries: a case study with B. thuringiensis Cry2A toxins

During evolution the creation of single crossover chimeras between duplicated paralogous genes is a known process for increasing diversity. Comparing the properties of homologously recombined chimeras with one or two crossovers is also an efficient strategy for analyzing relationships between sequence variation and function. However, no well-developed in vitro method has been established to create single-crossover libraries. Here we present an in vitro template-change polymerase change reaction that has been developed to enable the production of such libraries. We applied the method to two closely related toxin genes from B. thuringiensis and created chimeras with differing properties that can help us understand how these toxins are able to differentiate between insect species

Photometric variability in the open cluster M67 I. Cluster members detected in X-rays

We study photometric variability among the optical counterparts of X-ray sources in the old open cluster M67. The two puzzling binaries below the giant branch are both variables: for S1113 the photometric period is compatible with the orbital period, S1063 either varies on a period longer than the orbital period, or does not vary periodically. For the spectroscopic binaries S999, S1070 and S1077 the photometric and orbital periods are similar. Another new periodic variable is the main-sequence star S1112, not known to be a binary. An increase of the photometric period in the WUMa system S1282 (AHCnc) is in agreement with a previously reported trend. Six of the eight variables we detected are binaries with orbital periods of 10 days or less and equal photometric and orbital periods. This confirms the interpretation that their X-ray emission arises in the coronae of tidally locked magnetically active stars. No variability was found for the binaries with orbital periods longer than 40 days; their X-ray emission remains to be explained.Comment: 11 pages, accepted for publication in A&

The KNee OsteoArthritis Prediction (KNOAP2020) challenge:An image analysis challenge to predict incident symptomatic radiographic knee osteoarthritis from MRI and X-ray images

Objectives: The KNee OsteoArthritis Prediction (KNOAP2020) challenge was organized to objectively compare methods for the prediction of incident symptomatic radiographic knee osteoarthritis within 78 months on a test set with blinded ground truth. Design: The challenge participants were free to use any available data sources to train their models. A test set of 423 knees from the Prevention of Knee Osteoarthritis in Overweight Females (PROOF) study consisting of magnetic resonance imaging (MRI) and X-ray image data along with clinical risk factors at baseline was made available to all challenge participants. The ground truth outcomes, i.e., which knees developed incident symptomatic radiographic knee osteoarthritis (according to the combined ACR criteria) within 78 months, were not provided to the participants. To assess the performance of the submitted models, we used the area under the receiver operating characteristic curve (ROCAUC) and balanced accuracy (BACC). Results: Seven teams submitted 23 entries in total. A majority of the algorithms were trained on data from the Osteoarthritis Initiative. The model with the highest ROCAUC (0.64 (95% confidence interval (CI): 0.57–0.70)) used deep learning to extract information from X-ray images combined with clinical variables. The model with the highest BACC (0.59 (95% CI: 0.52–0.65)) ensembled three different models that used automatically extracted X-ray and MRI features along with clinical variables. Conclusion: The KNOAP2020 challenge established a benchmark for predicting incident symptomatic radiographic knee osteoarthritis. Accurate prediction of incident symptomatic radiographic knee osteoarthritis is a complex and still unsolved problem requiring additional investigation.</p

Excessive HDAC activation is critical for neurodegeneration in the rd1 mouse

Inherited retinal degenerations, collectively termed retinitis pigmentosa (RP), constitute one of the leading causes of blindness in the developed world. RP is at present untreatable and the underlying neurodegenerative mechanisms are unknown, even though the genetic causes are often established. Acetylation and deacetylation of histones, carried out by histone acetyltransferases (HATs) and histone deacetylases (HDACs), respectively, affects cellular division, differentiation, death and survival. We found acetylation of histones and probably other proteins to be dramatically reduced in degenerating photoreceptors in the rd1 human homologous mouse model for RP. Using a custom developed in situ HDAC activity assay, we show that overactivation of HDAC classes I/II temporally precedes photoreceptor degeneration. Moreover, pharmacological inhibition of HDACs I/II activity in rd1 organotypic retinal explants decreased activity of poly-ADP-ribose-polymerase and strongly reduced photoreceptor cell death. These findings highlight the importance of protein acetylation for photoreceptor cell death and survival and propose certain HDAC classes as novel targets for the pharmacological intervention in RP

Dominant Negative Mutants of Bacillus thuringiensis Cry1Ab Toxin Function as Anti-Toxins: Demonstration of the Role of Oligomerization in Toxicity

BACKGROUND:Bacillus thuringiensis Cry toxins, that are used worldwide in insect control, kill insects by a mechanism that depends on their ability to form oligomeric pores that insert into the insect-midgut cells. These toxins are being used worldwide in transgenic plants or spray to control insect pests in agriculture. However, a major concern has been the possible effects of these insecticidal proteins on non-target organisms mainly in ecosystems adjacent to agricultural fields. METHODOLOGY/PRINCIPAL FINDINGS:We isolated and characterized 11 non-toxic mutants of Cry1Ab toxin affected in different steps of the mechanism of action namely binding to receptors, oligomerization and pore-formation. These mutant toxins were analyzed for their capacity to block wild type toxin activity, presenting a dominant negative phenotype. The dominant negative phenotype was analyzed at two levels, in vivo by toxicity bioassays against susceptible Manduca sexta larvae and in vitro by pore formation activity in black lipid bilayers. We demonstrate that some mutations located in helix alpha-4 completely block the wild type toxin activity at sub-stoichiometric level confirming a dominant negative phenotype, thereby functioning as potent antitoxins. CONCLUSIONS/SIGNIFICANCE:This is the first reported case of a Cry toxin dominant inhibitor. These data demonstrate that oligomerization is a fundamental step in Cry toxin action and represent a potential mechanism to protect special ecosystems from the possible effect of Cry toxins on non-target organisms

Perinatal asphyxia: current status and approaches towards neuroprotective strategies, with focus on sentinel proteins

Delivery is a stressful and risky event menacing the newborn. The mother-dependent respiration has to be replaced by autonomous pulmonary breathing immediately after delivery. If delayed, it may lead to deficient oxygen supply compromising survival and development of the central nervous system. Lack of oxygen availability gives rise to depletion of NAD+ tissue stores, decrease of ATP formation, weakening of the electron transport pump and anaerobic metabolism and acidosis, leading necessarily to death if oxygenation is not promptly re-established. Re-oxygenation triggers a cascade of compensatory biochemical events to restore function, which may be accompanied by improper homeostasis and oxidative stress. Consequences may be incomplete recovery, or excess reactions that worsen the biological outcome by disturbed metabolism and/or imbalance produced by over-expression of alternative metabolic pathways. Perinatal asphyxia has been associated with severe neurological and psychiatric sequelae with delayed clinical onset. No specific treatments have yet been established. In the clinical setting, after resuscitation of an infant with birth asphyxia, the emphasis is on supportive therapy. Several interventions have been proposed to attenuate secondary neuronal injuries elicited by asphyxia, including hypothermia. Although promising, the clinical efficacy of hypothermia has not been fully demonstrated. It is evident that new approaches are warranted. The purpose of this review is to discuss the concept of sentinel proteins as targets for neuroprotection. Several sentinel proteins have been described to protect the integrity of the genome (e.g. PARP-1; XRCC1; DNA ligase IIIα; DNA polymerase β, ERCC2, DNA-dependent protein kinases). They act by eliciting metabolic cascades leading to (i) activation of cell survival and neurotrophic pathways; (ii) early and delayed programmed cell death, and (iii) promotion of cell proliferation, differentiation, neuritogenesis and synaptogenesis. It is proposed that sentinel proteins can be used as markers for characterising long-term effects of perinatal asphyxia, and as targets for novel therapeutic development and innovative strategies for neonatal care