Purpureotin: a novel di-dimeric C-type lectin-like protein from Trimeresurus purpureomaculatus venom is stabilized by noncovalent interactions

Purpureotin, a novel di-dimeric C-type lectin-like protein (CLP) from Trimeresurus purpureomaculatus, was purified and sequenced. While its native molecular mass was determined to be 63kDa, purpureotin showed a single band of 30kDa on nonreducing SDS-PAGE and two polypeptide chains (16.0 and 14.5kDa) under reducing condition. These results were subsequently confirmed by mass spectrometric analyses. Based on these results, we postulate that purpureotin is a dimer of the alpha,beta-heterodimer which is held together by noncovalent interactions. Molecular modeling studies indicate that a dimer of alpha,beta-heterodimers can be formed where the alpha chains are held together by electrostatic charges and beta chains via hydrophobic interactions. Functionally, purpureotin induced platelet aggregation without any cofactor in a dose-dependent manner. However, the platelet aggregation effect was blocked by echicetin. Therefore, purpureotin is assumed to be a GPIb-binding protein which binds to the same or a closely related GPIb site on platelets as echicetin.     

Hypoxia augments TNF-alpha-mediated endothelin-1 release and cell proliferation in human optic nerve head astrocytes

The effect of hypoxia (24 h) on TNF-alpha-mediated release of endothelin-1 (ET-1) from human optic nerve head astrocytes (hONAs) and TNF-alpha- and ET-1-induced hONA proliferation was determined. ET-1 synthesis and release was quantitated using ELISA while TNF-alpha (10 nM)- and ET-1 (100 nM)-mediated hONA proliferation was assessed by CellTiter 96 aqueous one-solution cell proliferation assay, respectively. hONAs appeared to be more rounded with fewer processes following 24 h hypoxia compared to thodr seen in normoxia. Hypoxia enhanced TNF-alpha-mediated ET-1 synthesis and release (by 5-fold) and also significantly increased TNF-alpha- and ET-1-mediated hONA proliferation. PD142893 (1 microM), an ET(A/B) receptor antagonist, blocked ET-1-mediated hONA proliferation both under normoxia and hypoxia, while doing so only under normoxia following TNF-alpha treatment. Also, U0126 (10 microM; an upstream ERK1/2 inhibitor) completely blocked agonist-induced hONA proliferation in normoxia and partially blocked the same in hypoxia. These results demonstrate for the first time that hONAs secrete ET-1 and that TNF-alpha and hypoxia can regulate its levels. Moreover, hypoxia augments the proliferative responses of hONAs to TNF-alpha and ET-1. These agonist-mediated effects following hypoxia could contribute to astroglial activation as seen in glaucomatous optic nerve heads.     

A flexible multi-dimensional QoS performance measure framework for distributed heterogeneous systems

When users’ tasks in a distributed heterogeneous computing environment (e.g., cluster of heterogeneous computers) are allocated resources, the total demand placed on some system resources by the tasks, for a given interval of time, may exceed the availability of those resources. In such a case, some tasks may receive degraded service or be dropped from the system. One part of a measure to quantify the success of a resource management system (RMS) in such a distributed environment is the collective value of the tasks completed during an interval of time, as perceived by the user, application, or policy maker. The Flexible Integrated System Capability (FISC) measure presented here is a measure for quantifying this collective value. The FISC measure is a flexible multi-dimensional measure such that any task attribute can be inserted and may include priorities, versions of a task or data, deadlines, situational mode, security, application- and domain-specific QoS, and task dependencies. For an environment where it is important to investigate how well data communication requests are satisfied, the data communication request satisfied can be the basis of the FISC measure instead of tasks completed. The motivation behind the FISC measure is to determine the performance of resource management schemes if tasks have multiple attributes that needs to be satisfied. The goal of this measure is to compare the results of different resource management heuristics that are trying to achieve the same performance objective but with different approaches. This research was supported by the DARPA/ITO Quorum Program, by the DARPA/ISO BADD Program and the Office of Naval Research under ONR grant number N00014-97-1-0804, by the DARPA/ITO AICE program under contract numbers DABT63-99-C-0010 and DABT63-99-C-0012, and by the Colorado State University George T. Abell Endowment. Intel and Microsoft donated some of the equipment used in this research. Jong-Kook Kim is pursuing a Ph.D. degree from the School of Electrical and Computer Engineering at Purdue University (expected in August 2004). Jong-Kook received his M.S. degree in electrical and computer engineering from Purdue University in May 2000. He received his B.S. degree in electronic engineering from Korea University, Seoul, Korea in 1998. He has presented his work at several international conferences and has been a reviewer for numerous conferences and journals. His research interests include heterogeneous distributed computing, computer architecture, performance measure, resource management, evolutionary heuristics, and power-aware computing. He is a student member of the IEEE, IEEE Computer Society, and ACM. Debra Hensgen is a member of the Research and Evaluation Team at OpenTV in Mountain View, California. OpenTV produces middleware for set-top boxes in support of interactive television. She received her Ph.D. in the area of Distributed Operating Systems from the University of Kentucky. Prior to moving to private industry, as an Associate Professor in the systems area, she worked with students and colleagues to design and develop tools and systems for resource management, network re-routing algorithms and systems that preserve quality of service guarantees, and visualization tools for performance debugging of parallel and distributed systems. She has published numerous papers concerning her contributions to the Concurra toolkit for automatically generating safe, efficient concurrent code, the Graze parallel processing performance debugger, the SAAM path information base, and the SmartNet and MSHN Resource Management Systems. Taylor Kidd is currently a Software Architect for Vidiom Systems in Portland Oregon. His current work involves the writing of multi-company industrial specifications and the architecting of software systems for the digital cable television industry. He has been involved in the establishment of international specifications for digital interactive television in both Europe and in the US. Prior to his current position, Dr. Kidd has been a researcher for the US Navy as well as an Associate Professor at the Naval Postgraduate School. Dr Kidd received his Ph.D. in Electrical Engineering in 1991 from the University of California, San Diego. H. J. Siegel was appointed the George T. Abell Endowed Chair Distinguished Professor of Electrical and Computer Engineering at Colorado State University (CSU) in August 2001, where he is also a Professor of Computer Science. In December 2002, he became the first Director of the CSU Information Science and Technology Center (ISTeC). ISTeC is a university-wide organization for promoting, facilitating, and enhancing CSU’s research, education, and outreach activities pertaining to the design and innovative application of computer, communication, and information systems. From 1976 to 2001, he was a professor at Purdue University. He received two BS degrees from MIT, and the MA, MSE, and PhD degrees from Princeton University. His research interests include parallel and distributed computing, heterogeneous computing, robust computing systems, parallel algorithms, parallel machine interconnection networks, and reconfigurable parallel computer systems. He has co-authored over 300 published papers on parallel and distributed computing and communication, is an IEEE Fellow, is an ACM Fellow, was a Coeditor-in-Chief of the Journal of Parallel and Distributed Computing, and was on the Editorial Boards of both the IEEE Transactions on Parallel and Distributed Systems and the IEEE Transactions on Computers. He was Program Chair/Co-Chair of three major international conferences, General Chair/Co-Chair of four international conferences, and Chair/Co-Chair of five workshops. He has been an international keynote speaker and tutorial lecturer, and has consulted for industry and government. David St. John is Chief Information Officer for WeatherFlow, Inc., a weather services company specializing in coastal weather observations and forecasts. He received a master’s degree in Engineering from the University of California, Irvine. He spent several years as the head of staff on the Management System for Heterogeneous Networks project in the Computer Science Department of the Naval Postgraduate School. His current relationship with cluster computing is as a user of the Regional Atmospheric Modeling System (RAMS), a numerical weather model developed at Colorado State University. WeatherFlow runs RAMS operationally on a Linux-based cluster. Cynthia Irvine is a Professor of Computer Science at the Naval Postgraduate School in Monterey, California. She received her Ph.D. from Case Western Reserve University and her B.A. in Physics from Rice University. She joined the faculty of the Naval Postgraduate School in 1994. Previously she worked in industry on the development of high assurance secure systems. In 2001, Dr. Irvine received the Naval Information Assurance Award. Dr. Irvine is the Director of the Center for Information Systems Security Studies and Research at the Naval Postgraduate School. She has served on special panels for NSF, DARPA, and OSD. In the area of computer security education Dr. Irvine has most recently served as the general chair of the Third World Conference on Information Security Education and the Fifth Workshop on Education in Computer Security. She co-chaired the NSF workshop on Cyber-security Workforce Needs Assessment and Educational Innovation and was a participant in the Computing Research Association/NSF sponsored Grand Challenges in Information Assurance meeting. She is a member of the editorial board of the Journal of Information Warfare and has served as a reviewer and/or program committee member of a variety of security related conferences. She has written over 100 papers and articles and has supervised the work of over 80 students. Professor Irvine is a member of the ACM, the AAS, a life member of the ASP, and a Senior Member of the IEEE. Timothy E. Levin is a Research Associate Professor at the Naval Postgraduate School. He has spent over 18 years working in the design, development, evaluation, and verification of secure computer systems, including operating systems, databases and networks. His current research interests include high assurance system design and analysis, development of models and methods for the dynamic selection of QoS security attributes, and the application of formal methods to the development of secure computer systems. Viktor K. Prasanna received his BS in Electronics Engineering from the Bangalore University and his MS from the School of Automation, Indian Institute of Science. He obtained his Ph.D. in Computer Science from the Pennsylvania State University in 1983. Currently, he is a Professor in the Department of Electrical Engineering as well as in the Department of Computer Science at the University of Southern California, Los Angeles. He is also an associate member of the Center for Applied Mathematical Sciences (CAMS) at USC. He served as the Division Director for the Computer Engineering Division during 1994–98. His research interests include parallel and distributed systems, embedded systems, configurable architectures and high performance computing. Dr. Prasanna has published extensively and consulted for industries in the above areas. He has served on the organizing committees of several international meetings in VLSI computations, parallel computation, and high performance computing. He is the Steering Co-chair of the International Parallel and Distributed Processing Symposium [merged IEEE International Parallel Processing Symposium (IPPS) and the Symposium on Parallel and Distributed Processing (SPDP)] and is the Steering Chair of the International Conference on High Performance Computing(HiPC). He serves on the editorial boards of the Journal of Parallel and Distributed Computing and the Proceedings of the IEEE. He is the Editor-in-Chief of the IEEE Transactions on Computers. He was the founding Chair of the IEEE Computer Society Technical Committee on Parallel Processing. He is a Fellow of the IEEE. Richard F. Freund is the originator of GridIQ’s network scheduling concepts that arose from mathematical and computing approaches he developed for the Department of Defense in the early 1980’s. Dr. Freund has over twenty-five years experience in computational mathematics, algorithm design, high performance computing, distributed computing, network planning, and heterogeneous scheduling. Since 1989, Dr. Freund has published over 45 journal articles in these fields. He has also been an editor of special editions of IEEE Computer and the Journal of Parallel and Distributed Computing. In addition, he is a founder of the Heterogeneous Computing Workshop, held annually in conjunction with the International Parallel Processing Symposium. Dr. Freund is the recipient of many awards, which includes the prestigious Department of Defense Meritorious Civilian Service Award in 1984 and the Lauritsen-Bennet Award from the Space and Naval Warfare Systems Command in San Diego, California.     

Increase of homologous recombination frequency in vascular tissue of Arabidopsis plants exposed to salt stress

Here we analyzed the influence of salt stress on plant genome stability. Homologous recombination events were detected in transgenic Arabidopsis plants that carried in their genome a beta-glucuronidase recombination marker. Recombination events were scored as blue sectors using a stereo microscope. Exposure to 50 mM salt resulted in a 3.0-fold increase in recombination frequency. To analyze the organ and tissue specificity of recombination events, we examined cross-sections of leaves, stems and roots. We found that nearly 30% of recombination events in plants grown under normal conditions and nearly 50% of events in plants grown on salt were undetected by the conventional method. Most of the recombination events represented a cluster/group of cells (12 on average), although events with single cells were also detected. Recombination events were very frequent in leaf mesophyll cells. On average, individual recombination events located on leaves contained more cells than events located on roots or stems. Analysis of recombination events in cross-sectioned tissue of salt-treated plants revealed a shift in the distribution of recombination events towards the vascular tissue. We discuss the significance of the finding for plant stress physiology.     

Development of alternative support system for viable count of cyanobacteria by most probable number method

A technique was developed to evaluate alternative support systems to test tubes used in the standard most probable number technique, for simultaneous isolation and enumeration of cyanobacteria. Five different support systems were tested for their suitability in terms of accuracy, sensitivity, economics and ease of handling. PCR plates with 96 wells and carrying capacity of 300 microL per well were found to be most sensitive, besides being cost- and time-effective. This technique can also be useful for isolation of cyanobacteria, due to immobilization of colonies in the gel matrix and storage of samples at room temperature, without loss of viability for 5-6 weeks. This technique can help to process large sample size with ease--both for enumeration and isolation and can be extended for enumeration of other microorganisms from diverse sources.     

Bioprospecting microalgae as potential sources of "green energy"--challenges and perspectives (review)

Microalgae and cyanobacteria are potential foods, feeds, sources of high-value bioactive molecules and biofuels, and find tremendous applications in bioremediation and agriculture. Although few efforts have been undertaken to index the microalgal germplasm available in terms of lipid content, information on suitability of strains for mass multiplication and advances in development of methods for extraction and generating biofuel are scarce. Our review summarizes the potential of microalgae, latest developments in the field and analyzes the "pitfalls" in oversimplification of their promise in the years to come. Microalgae represent "green gold mines" for generating energy; however, the path to success is long and winding and needs tremendous and concerted efforts from science and industry, besides political will and social acceptance for overcoming the limitations. The major advantages of second generation biofuels based on microalgal systems, include their higher photon conversion efficiency, growth all around the year, even in wastewaters, and production of environment friendly biodegradable biofuels.     

Argonaute identity defines the length of mature mammalian microRNAs

MicroRNAs (miRNAs) are 19- to 25-nt-long non-coding RNAs that regulate gene expression by base-pairing with target mRNAs and reducing their stability or translational efficiency. Mammalian miRNAs function in association with four closely related Argonaute proteins, AGO1-4. All four proteins contain the PAZ and the MID domains interacting with the miRNA 3' and 5' termini, respectively, as well as the PIWI domain comprising an mRNA 'slicing' activity in the case of AGO2 but not AGO1, AGO3 and AGO4. However, the slicing mode of the miRNA-programmed AGO2 is rarely realized in vivo and the four Argonautes are thought to play largely overlapping roles in the mammalian miRNA pathway. Here, we show that the average length of many miRNAs is diminished during nervous system development as a result of progressive shortening of the miRNA 3' ends. We link this modification with an increase in the fractional abundance of Ago2 in the adult brain and identify a specific structural motif within the PAZ domain that enables efficient trimming of miRNAs associated with this but not the other three Argonautes. Taken together, our data suggest that mammalian Argonautes may define the length and possibly biological activity of mature mammalian miRNAs in a developmentally controlled manner.     

NALP-3 inflammasome silencing attenuates ceramide-induced transepithelial permeability

The hallmark of acute lung injury (ALI) is the influx of proinflammatory cytokines into lung tissue and alveolar permeability that ultimately leads to pulmonary edema. However, the mechanisms involved in inflammatory cytokine production and alveolar permeability are unclear. Recent studies suggest that excessive production of ceramide has clinical relevance as a mediator of pulmonary edema and ALI. Our earlier studies indicate that the activation of inflammasome promotes the processing and secretion of proinflammatory cytokines and causes alveolar permeability in ALI. However, the role of ceramide in inflammasome activation and the underlying mechanism in relation to alveolar permeability is not known. We hypothesized that ceramide activates the inflammasome and causes inflammatory cytokine production and alveolar epithelial permeability. To test this hypothesis, we analyzed the lung ceramide levels during hyperoxic ALI in mice. The effect of ceramide on activation of inflammasome and production of inflammatory cytokine was assessed in primary mouse alveolar macrophages and THP-1 cells. Alveolar transepithelial permeability was determined in alveolar epithelial type-II cells (AT-II) and THP-1 co-cultures. Our results reveal that ceramide causes inflammasome activation, induction of caspase-1, IL-1β cleavage, and release of proinflammatory cytokines. In addition, ceramide further induces alveolar epithelial permeability. Short-hairpin RNA silencing of inflammasome components abrogated ceramide-induced secretion of proinflammatory cytokines in vitro. Inflammasome silencing abolishes ceramide-induced alveolar epithelial permeability in AT-II. Collectively, our results demonstrate for the first time that ceramide-induced secretion of proinflammatory cytokines and alveolar epithelial permeability occurs though inflammasome activation.