Interaction of Cholesterol with Perfringolysin O: What Have We Learned from Functional Analysis?

Cholesterol-dependent cytolysins (CDCs) constitute a family of pore-forming toxins secreted by Gram-positive bacteria. These toxins form transmembrane pores by inserting a large β-barrel into cholesterol-containing membranes. Cholesterol is absolutely required for pore-formation. For most CDCs, binding to cholesterol triggers conformational changes that lead to oligomerization and end in pore-formation. Perfringolysin O (PFO), secreted by Clostridium perfringens, is the prototype for the CDCs. The molecular mechanisms by which cholesterol regulates the cytolytic activity of the CDCs are not fully understood. In particular, the location of the binding site for cholesterol has remained elusive. We have summarized here the current body of knowledge on the CDCs-cholesterol interaction, with focus on PFO. We have employed sterols in aqueous solution to identify structural elements in the cholesterol molecule that are critical for its interaction with PFO. In the absence of high-resolution structural information, site-directed mutagenesis data combined with binding studies performed with different sterols, and molecular modeling are beginning to shed light on this interaction

A Dynamic Risk-Based Access Control Approach: Model and Implementation

Access control (AC) refers to mechanisms and policies that restrict access to resources, thus regulating access to physical or virtual resources of an information system. AC approaches are used to represent these mechanisms and policies by which users are granted access and specific access privileges to the resources or information of the system for which AC is provided. Traditional AC approaches encompass a variety of widely used approaches, including attribute-based access control (ABAC), mandatory access control (MAC), discretionary access control (DAC) and role-based access control (RBAC). Emerging AC approaches include risk adaptive access control (RAdAC), an approach that suggests that AC can adapt depending on specific situations. However, traditional and emerging AC approaches rely on static pre-defined risk mitigation tasks and do not support the adaptation of an AC risk mitigation process (RMP). There are no provided mechanisms and automated support that allow AC approaches to construct RMPs and to adapt to provide more flexible, custom-tailored responses to specific situations in order to minimize risks. Further, although existing AC approaches can operate in several knowledge domains at once, they do not explicitly take into account the relationships among risks related to different dimensions, e.g., security, productivity. In addition, although in the real world, risks accumulate over time, existing AC approaches do not appropriately provide means for risk resolution in situations in which risks accumulate as different, dangerous tasks impact risk measures. This thesis presents the definition, the implementation, and the application through two case studies of a novel AC risk-mitigation approach that combines dynamic RMP construction and risk assessment extended to include forecasting based on multiple risk-related utilities and events; provides support for a dynamic risk assessment that depends on one or multiple risk dimensions (e.g., security and productivity); offers cumulative risk assessment in which each action of interest can impact the risk-related utilities in a dynamic way; and presents an implementation of an adaptive simulation method based on risk-related utilities and events

Drug Screening for Discovery of Broad-spectrum Agents for Soil-transmitted Nematodes

Soil-transmitted nematodes (STNs), namely hookworms, whipworms, and ascarids, are extremely common parasites, infecting 1-2 billion of the poorest people worldwide. Two benzimidazoles, albendazole and mebendazole, are currently used in STN mass drug administration, with many instances of low/reduced activity reported. New drugs against STNs are urgently needed. We tested various models for STN drug screening with the aim of identifying the most effective tactics for the discovery of potent, safe and broad-spectrum agents. We screened a 1280-compound library of approved drugs to completion against late larval/adult stages and egg/larval stages of both the human hookworm parasite Ancylostoma ceylanicum and the free-living nematode Caenorhabditis elegans, which is often used as a surrogate for STNs in screens. The quality of positives was further evaluated based on cheminformatics/data mining analyses and activity against evolutionarily distant Trichuris muris whipworm adults. From these data, two pairs of positives, sulconazole/econazole and pararosaniline/cetylpyridinium, predicted to target nematode CYP-450 and HSP-90 respectively, were prioritized for in vivo evaluation against A. ceylanicum infections in hamsters. One of these positives, pararosaniline, showed a significant impact on hookworm fecundity in vivo. Taken together, our results suggest that anthelmintic screening with A. ceylanicum larval stages is superior to C. elegans based on both reduced false negative rate and superior overall quality of actives. Our results also highlight two potentially important targets for the discovery of broad-spectrum human STN drugs

Coherent x-ray radiation induced by high-current breakdown on a ferrite surface

We observe that at the initial stage of a high-current discharge, a low-divergence short x-ray pulse ($\approx\thinspace$$0.5^{\circ}$, $\thinspace$$500$ eV) with the energy of $\sim$$21\mu$J is formed over a ferrite surface, which propagates parallel to the surface in the anode direction. The high directionality of the radiation points to its coherent nature. We propose that the radiation is due to the short-lived magnetization of the ferrite surface excited by a high-power electromagnetic pulse. The radiation is coherent due to the equivalent excitation conditions for all emitters. The excitation pulse and the radiation it generates move at the same speed ($\sim$$c$). Thereby, the emitted waves propagating parallel to the ferrite surface are phase-matched, providing the high radiant intensity of the radiation

Interrogating Plant Cell Culture Library for Novel Antimicrobial Agents

The Plant Cell Culture Library (PCCL) at UMass Amherst contains more than 2,200 live plant cell cultures, representing diverse plant species from around the world. The availability of this collection offers a rich resource for us to discover bioactive phytochemicals and uncover their mechanisms of action. Using data-mining surveys of bioactive plant extracts, I have organized subsets of PCCL cell lines that are likely to possess antifungal, antibacterial, antiviral, anthelmintic, anti-trypanosomal, or anticancer properties, which prove to be useful when deciding which species to screen first against a specific pathogen. Another distinct advantage of using the live plant cells in this research is the ability to stimulate the biosynthesis of pathogen-specific phytochemicals upon simulation of an attack (elicitation) by the microorganism in question. This could be accomplished by pathogen homogenates or plant hormones responsible for mounting defenses to infection. Over the past six months, I have been working to optimize elicitation, lysis, and extraction conditions for obtaining high-throughput screening materials to be used against variable pathogens. Equipped with crude extracts from appropriately elicited cells, I am collaborating with a multidisciplinary team of UMass scientists to develop and implement high-throughput screening protocols for profiling a large number of plant-derived materials against various pathogens. Recently, I have screened a small pool (40) of extracts derived from cell lines with predicted anti-fungal properties against the highly resistant strain of fungus Fusarium oxysporum, one of the causal agents of an opportunistic infection often seen in immunocompromised patients known as fusariosis. Gratifyingly, I have found several plant species that produced specialized metabolites with better antifungal activity than the leading antibiotic against F. oxysporum, Amphotericin B, validating this line of antimicrobial research. We are also actively reaching out to other academic labs partners to form partnerships in diverse antimicrobial research venues

Screening studies of POP levels in bottom sediments from selected lakes in the Paz watercourse

Appendix 5/15 of the publication "State of the environment in the Norwegian, Finnish and Russian border area 2007" (The Finnish Environment 6/2007)

Screening studies of POP levels in fish from selected lakes in the Paz watercourse

Appendix 8/15 of the publication "State of the environment in the Norwegian, Finnish and Russian border area 2007" (The Finnish Environment 6/2007)

Evaluation of AaDOP2 Receptor Antagonists Reveals Antidepressants and Antipsychotics as Novel Lead Molecules for Control of the Yellow Fever Mosquito, Aedes aegypti s

ABSTRACT The yellow fever mosquito, Aedes aegypti, vectors disease-causing agents that adversely affect human health, most notably the viruses causing dengue and yellow fever. The efficacy of current mosquito control programs is challenged by the emergence of insecticideresistant mosquito populations, suggesting an urgent need for the development of chemical insecticides with new mechanisms of action. One recently identified potential insecticide target is the A. aegypti D 1 -like dopamine receptor, AaDOP2. The focus of the present study was to evaluate AaDOP2 antagonism both in vitro and in vivo using assay technologies with increased throughput. The in vitro assays revealed AaDOP2 antagonism by four distinct chemical scaffolds from tricyclic antidepressant or antipsychotic chemical classes, and elucidated several structure-activity relationship trends that contributed to enhanced antagonist potency, including lipophilicity, halide substitution on the tricyclic core, and conformational rigidity. Six compounds displayed previously unparalleled potency for in vitro AaDOP2 antagonism, and among these, asenapine, methiothepin, and cis-(Z)-flupenthixol displayed subnanomolar IC 50 values and caused rapid toxicity to A. aegypti larvae and/or adults in vivo. Our study revealed a significant correlation between in vitro potency for AaDOP2 antagonism and in vivo toxicity, suggesting viability of AaDOP2 as an insecticidal target. Taken together, this study expanded the repertoire of known AaDOP2 antagonists, enhanced our understanding of AaDOP2 pharmacology, provided further support for rational targeting of AaDOP2, and demonstrated the utility of efficiency-enhancing in vitro and in vivo assay technologies within our genome-to-lead pipeline for the discovery of next-generation insecticides

Dosimetry for boron neutron capture therapy developed and verified at the accelerator based neutron source VITA

The method of boron neutron capture therapy for malignant tumors, proposed in 1936, is beginning to enter clinical practice. The development of dosimetry tools for characterization of therapeutic mixed neutron-photon beam and assessing the patient’s response to treatment is becoming relevant. In this work, a number of dosimetric techniques have been developed: a compact neutron detector with a pair of cast scintillators, one of which is enriched with boron, to measure the boron dose and the γ-ray dose; cell dosimeter for measuring the sum of the equivalent dose of fast neutrons and the equivalent nitrogen dose; prompt γ-ray spectroscopy for in situ measurement of boron dose in real time; epithermal neutron flux monitor for measuring the epithermal neutron flux. Their verification carried out on the accelerator based neutron source VITA showed that they can become convenient and reliable tools for characterization of neutron beam and assessing the patient’s response to treatment