Ricin Toxicokinetics and Its Sensitive Detection in Mouse Sera or Feces Using Immuno-PCR

Ricin (also called RCA-II or RCA(60)), one of the most potent toxins and documented bioweapons, is derived from castor beans of Ricinus communis. Several in vitro methods have been designed for ricin detection in complex food matrices in the event of intentional contamination. Recently, a novel Immuno-PCR (IPCR) assay was developed with a limit of detection of 10 fg/ml in a buffer matrix and about 10-1000-fold greater sensitivity than other methods in various food matrices.In order to devise a better diagnostic test for ricin, the IPCR assay was adapted for the detection of ricin in biological samples collected from mice after intoxication. The limit of detection in both mouse sera and feces was as low as 1 pg/ml. Using the mouse intravenous (iv) model for ricin intoxication, a biphasic half-life of ricin, with a rapid t(1/2)α of 4 min and a slower t(1/2)β of 86 min were observed. The molecular biodistribution time for ricin following oral ingestion was estimated using an antibody neutralization assay. Ricin was detected in the blood stream starting at approximately 6-7 h post- oral intoxication. Whole animal histopathological analysis was performed on mice treated orally or systemically with ricin. Severe lesions were observed in the pancreas, spleen and intestinal mesenteric lymph nodes, but no severe pathology in other major organs was observed.The determination of in vivo toxicokinetics and pathological effects of ricin following systemic and oral intoxication provide a better understanding of the etiology of intoxication and will help in the future design of more effective diagnostic and therapeutic methods

Real-Time Cytotoxicity Assay for Rapid and Sensitive Detection of Ricin from Complex Matrices

BACKGROUND: In the context of a potential bioterrorist attack sensitive and fast detection of functionally active toxins such as ricin from complex matrices is necessary to be able to start timely countermeasures. One of the functional detection methods currently available for ricin is the endpoint cytotoxicity assay, which suffers from a number of technical deficits. METHODOLOGY/FINDINGS: This work describes a novel online cytotoxicity assay for the detection of active ricin and Ricinus communis agglutinin, that is based on a real-time cell electronic sensing system and impedance measurement. Characteristic growth parameters of Vero cells were monitored online and used as standardized viability control. Upon incubation with toxin the cell status and the cytotoxic effect were visualized using a characteristic cell index-time profile. For ricin, tested in concentrations of 0.06 ng/mL or above, a concentration-dependent decrease of cell index correlating with cytotoxicity was recorded between 3.5 h and 60 h. For ricin, sensitive detection was determined after 24 h, with an IC50 of 0.4 ng/mL (for agglutinin, an IC50 of 30 ng/mL was observed). Using functionally blocking antibodies, the specificity for ricin and agglutinin was shown. For detection from complex matrices, ricin was spiked into several food matrices, and an IC50 ranging from 5.6 to 200 ng/mL was observed. Additionally, the assay proved to be useful in detecting active ricin in environmental sample materials, as shown for organic fertilizer containing R. communis material. CONCLUSIONS/SIGNIFICANCE: The cell-electrode impedance measurement provides a sensitive online detection method for biologically active cytotoxins such as ricin. As the cell status is monitored online, the assay can be standardized more efficiently than previous approaches based on endpoint measurement. More importantly, the real-time cytotoxicity assay provides a fast and easy tool to detect active ricin in complex sample matrices

Evidence for widespread hydrated minerals on asteroid (101955) Bennu

Early spectral data from the Origins, Spectral Interpretation, Resource Identification, and Security-Regolith Explorer (OSIRIS-REx) mission reveal evidence for abundant hydrated minerals on the surface of near-Earth asteroid (101955) Bennu in the form of a near-infrared absorption near 2.7 µm and thermal infrared spectral features that are most similar to those of aqueously altered CM-type carbonaceous chondrites. We observe these spectral features across the surface of Bennu, and there is no evidence of substantial rotational variability at the spatial scales of tens to hundreds of metres observed to date. In the visible and near-infrared (0.4 to 2.4 µm) Bennu’s spectrum appears featureless and with a blue (negative) slope, confirming previous ground-based observations. Bennu may represent a class of objects that could have brought volatiles and organic chemistry to Earth

The dynamic geophysical environment of (101955) Bennu based on OSIRIS-REx measurements

The top-shaped morphology characteristic of asteroid (101955) Bennu, often found among fast-spinning asteroids and binary asteroid primaries, may have contributed substantially to binary asteroid formation. Yet a detailed geophysical analysis of this morphology for a fast-spinning asteroid has not been possible prior to the Origins, Spectral Interpretation, Resource Identification, and Security-Regolith Explorer (OSIRIS-REx) mission. Combining the measured Bennu mass and shape obtained during the Preliminary Survey phase of the OSIRIS-REx mission, we find a notable transition in Bennu’s surface slopes within its rotational Roche lobe, defined as the region where material is energetically trapped to the surface. As the intersection of the rotational Roche lobe with Bennu’s surface has been most recently migrating towards its equator (given Bennu’s increasing spin rate), we infer that Bennu’s surface slopes have been changing across its surface within the last million years. We also find evidence for substantial density heterogeneity within this body, suggesting that its interior is a mixture of voids and boulders. The presence of such heterogeneity and Bennu’s top shape are consistent with spin-induced failure at some point in its past, although the manner of its failure cannot yet be determined. Future measurements by the OSIRIS-REx spacecraft will provide insight into and may resolve questions regarding the formation and evolution of Bennu’s top-shape morphology and its link to the formation of binary asteroids

Evolutionary Hypotheses and Moral Skepticism

Proponents of evolutionary debunking arguments aim to show that certain genealogical explanations of our moral faculties, if true, undermine our claim to moral knowledge. Criticisms of these arguments generally take the debunker’s genealogical explanation for granted. The task of the anti-debunker is thought to be that of reconciling the (supposed) truth of this hypothesis with moral knowledge. In this paper, I shift the critical focus instead to the debunker’s empirical hypothesis and argue that the skeptical strength of an evolutionary debunking argument is dependent upon the evidence for that hypothesis—evidence which, upon further inspection, proves far from compelling. Following that, however, I suggest that the same considerations which spell trouble for the empirical hypotheses of traditional debunking arguments can also be taken to give rise to an alternative—and better supported—style of debunking argument

Role of 5-HT 1A receptors in antidepressant-like effect of dichloromethane fraction of Kielmeyera coriacea in rats subjected to the forced swim test

Objective : We examined the involvement of 5-HT neurotransmission on the antidepressant-like effect of the dichloromethane (DcM) fraction of an extract from Kielmeyera coriacea stems. Materials and Methods : Male Wistar rats treated chronically (45 days, gavage) with the DcM fraction received an intradorsal raphe nucleus (DRN) microinjection of saline or 5-HT 1A receptor ligands and were evaluated in the forced swimming test (FST) and in the open-field test (OFT). Results : The DcM fraction (5.0 mg/kg) reduced immobility time in the FST without altering locomotion in the OFT. IntraDRN microinjection of the 5-HT 1A receptor agonist, (+)-8-OH-DPAT (0.10; 0.20 or 0.33 μg) increased immobility time and reduced locomotion at the higher dose whereas the 5-HT1A antagonists, (-)-pindolol (0.10; 0.20 or 0.40 μg) or WAY100635 (0.11; 0.22 or 0.43 μg) did not produce any effect in the behavioral tests. IntraDRN (+)-8-OH-DPAT (0.20 or 0.33 μg) in rats treated with the DcM fraction (5.0 mg/kg) blocked the changes in the immobility time or in locomotion produced by each drug. Intra-DRN (-)-pindolol (0.10 μg) or WAY100635 (0.43 μg) in rats treated with a subactive dose of the DcM fraction (4.0 mg/kg) synergistically reduced immobility time in the FST. Conclusion : The DcM fraction of Kielmeyera coriacea produced an antidepressant-like effect in the FST and interacted with 5-HT 1A receptor ligands. Activation of 5-HT 1A receptors into DRN by (+) 8-OH-DPAT produced detectable changes in the FST or in the OFT