Distributed File System Virtualization Techniques Supporting On-Demand Virtual Machine Environments for Grid Computing

This paper presents a data management solution which allows fast Virtual Machine (VM) instantiation and efficient run-time execution to support VMs as execution environments in Grid computing. It is based on novel distributed file system virtualization techniques and is unique in that: (1) it provides on-demand cross-domain access to VM state for unmodified VM monitors; (2) it enables private file system channels for VM instantiation by secure tunneling and session-key based authentication; (3) it supports user-level and write-back disk caches, per-application caching policies and middleware-driven consistency models; and (4) it leverages application-specific meta-data associated with files to expedite data transfers. The paper reports on its performance in wide-area setups using VMware-based VMs. Results show that the solution delivers performance over 30% better than native NFS and with warm caches it can bring the application-perceived overheads below 10% compared to a local-disk setup. The solution also allows a VM with 1.6 GB virtual disk and 320 MB virtual memory to be cloned within 160 seconds for the first clone and within 25 seconds for subsequent clones. Ming Zhao is a PhD candidate in the department of Electrical and Computer Engineering and a member of the Advance Computing and Information Systems Laboratory, at University of Florida. He received the degrees of BE and ME from Tsinghua University. His research interests are in the areas of computer architecture, operating systems and distributed computing. Jian Zhang is a PhD student in the Department of Electrical and Computer Engineering at University of Florida and a member of the Advance Computing and Information Systems Laboratory (ACIS). Her research interest is in virtual machines and Grid computing. She is a member of the IEEE and the ACM. Renato J. Figueiredo received the B.S. and M.S. degrees in Electrical Engineering from the Universidade de Campinas in 1994 and 1995, respectively, and the Ph.D. degree in Electrical and Computer Engineering from Purdue University in 2001. From 2001 until 2002 he was on the faculty of the School of Electrical and Computer Engineering of Northwestern University at Evanston, Illinois. In 2002 he joined the Department of Electrical and Computer Engineering of the University of Florida as an Assistant Professor. His research interests are in the areas of computer architecture, operating systems, and distributed systems.     

Exploiting redundancy to boost performance in a RAID-10 style cluster-based file system

While aggregating the throughput of existing disks on cluster nodes is a cost-effective approach to alleviate the I/O bottleneck in cluster computing, this approach suffers from potential performance degradations due to contentions for shared resources on the same node between storage data processing and user task computation. This paper proposes to judiciously utilize the storage redundancy in the form of mirroring existed in a RAID-10 style file system to alleviate this performance degradation. More specifically, a heuristic scheduling algorithm is developed, motivated from the observations of a simple cluster configuration, to spatially schedule write operations on the nodes with less load among each mirroring pair. The duplication of modified data to the mirroring nodes is performed asynchronously in the background. The read performance is improved by two techniques: doubling the degree of parallelism and hot-spot skipping. A synthetic benchmark is used to evaluate these algorithms in a real cluster environment and the proposed algorithms are shown to be very effective in performance enhancement. Yifeng Zhu received his B.Sc. degree in Electrical Engineering in 1998 from Huazhong University of Science and Technology, Wuhan, China; the M.S. and Ph.D. degree in Computer Science from University of Nebraska – Lincoln in 2002 and 2005 respectively. He is an assistant professor in the Electrical and Computer Engineering department at University of Maine. His main research interests are cluster computing, grid computing, computer architecture and systems, and parallel I/O storage systems. Dr. Zhu is a Member of ACM, IEEE, the IEEE Computer Society, and the Francis Crowe Society. Hong Jiang received the B.Sc. degree in Computer Engineering in 1982 from Huazhong University of Science and Technology, Wuhan, China; the M.A.Sc. degree in Computer Engineering in 1987 from the University of Toronto, Toronto, Canada; and the PhD degree in Computer Science in 1991 from the Texas A&M University, College Station, Texas, USA. Since August 1991 he has been at the University of Nebraska-Lincoln, Lincoln, Nebraska, USA, where he is Professor and Vice Chair in the Department of Computer Science and Engineering. His present research interests are computer architecture, parallel/distributed computing, cluster and Grid computing, computer storage systems and parallel I/O, performance evaluation, real-time systems, middleware, and distributed systems for distance education. He has over 100 publications in major journals and international Conferences in these areas and his research has been supported by NSF, DOD and the State of Nebraska. Dr. Jiang is a Member of ACM, the IEEE Computer Society, and the ACM SIGARCH. Xiao Qin received the BS and MS degrees in computer science from Huazhong University of Science and Technology in 1992 and 1999, respectively. He received the PhD degree in computer science from the University of Nebraska-Lincoln in 2004. Currently, he is an assistant professor in the department of computer science at the New Mexico Institute of Mining and Technology. He had served as a subject area editor of IEEE Distributed System Online (2000–2001). His research interests are in parallel and distributed systems, storage systems, real-time computing, performance evaluation, and fault-tolerance. He is a member of the IEEE. Dan Feng received the Ph.D degree from Huazhong University of Science and Technology, Wuhan, China, in 1997. She is currently a professor of School of Computer, Huazhong University of Science and Technology, Wuhan, China. She is the principal scientist of the the National Grand Fundamental Research 973 Program of China “Research on the organization and key technologies of the Storage System on the next generation Internet.” Her research interests include computer architecture, storage system, parallel I/O, massive storage and performance evaluation. David Swanson received a Ph.D. in physical (computational) chemistry at the University of Nebraska-Lincoln (UNL) in 1995, after which he worked as an NSF-NATO postdoctoral fellow at the Technical University of Wroclaw, Poland, in 1996, and subsequently as a National Research Council Research Associate at the Naval Research Laboratory in Washington, DC, from 1997–1998. In 1999 he returned to UNL where he directs the Research Computing Facility and currently serves as an Assistant Research Professor in the Department of Computer Science and Engineering. The Office of Naval Research, the National Science Foundation, and the State of Nebraska have supported his research in areas such as large-scale scientific simulation and distributed systems.     

Polymeric and Compositional Properties of Novel Extracellular Microbial Polyglucosamine Biopolymer from New Strain of Citrobacter sp. BL-4

A novel polyglucosamine polymer, PGB-2, was produced extracellularly from a new strain Citrobacter sp. BL-4 using pH-stat fed batch cultivation. It was composed of 97.3% glucosamine and 2.7% rhamnose; its average molecular weight, solubility in 2% acetic acid and viscosity were 20 kDa, 5 g l⁻¹ and 2.9 cps, respectively. FT-IR and ¹H NMR spectra of PGB-2 revealed a close identity with chitosan from crab shells. Received 20 September 2005; Revisions requested 6 October 2005; Revisions received 16 November 2005; Accepted 16 November 2005     

Comparison of cell lines for stable production of fucose-negative antibodies with enhanced ADCC

Several methods have been described to enhance antibody-dependent cellular cytotoxicity (ADCC) using different host cells that produce antibody with reduced levels of fucose on their carbohydrates. We compared the suitability of these methods for the serum-free fed-batch production of antibody for clinical trials and commercial uses. Recombinant anti-human CD20 chimeric IgG1-producing clones were established from host-cells that have been shown to produce more than 90% fucose-negative antibody. The cell lines were a FUT8 (alpha-1,6-fucosyltransferase) knockout Chinese hamster ovary (CHO) cell line, Ms704, and two Lens culinaris agglutinin (LCA)-resistant cell lines, one derived from a variant CHO line, Lec13 and the other from a rat hybridoma cell line, YB2/0. The amount of fucose-negative antibody produced by Lec13 and YB2/0 significantly decreased with the culture. The increase in fucosylation was due to remaining synthesis of GDP-fucose via de novo pathway for the CHO line and the elevation of FUT8 expression by the YB2/0 cells. In contrast, Ms704 cells stably produced fucose-negative antibody with a consistent carbohydrate structure until the end of the culture. The productivity of the Ms704 cells reached 1.76 g/L with a specific production rate (SPR) of 29 pg/cell/day for 17 days in serum-free fed-batch culture using a 1 L spinner bioreactor. Our results demonstrate that FUT8 knockout has the essential characteristics of host cells for robust manufacture of fucose-negative therapeutic antibodies with enhanced ADCC.     

Event-driven time-optimal control for a class of discontinuous bioreactors

Discontinuous bioreactors may be further optimized for processing inhibitory substrates using a convenient fed-batch mode. To do so the filling rate must be controlled in such a way as to push the reaction rate to its maximum value, by increasing the substrate concentration just up to the point where inhibition begins. However, an exact optimal controller requires measuring several variables (e.g., substrate concentrations in the feed and in the tank) and also good model knowledge (e.g., yield and kinetic parameters), requirements rarely satisfied in real applications. An environmentally important case, that exemplifies all these handicaps, is toxicant wastewater treatment. There the lack of online practical pollutant sensors may allow unforeseen high shock loads to be fed to the bioreactor, causing biomass inhibition that slows down the treatment process and, in extreme cases, even renders the biological process useless. In this work an event-driven time-optimal control (ED-TOC) is proposed to circumvent these limitations. We show how to detect a "there is inhibition" event by using some computable function of the available measurements. This event drives the ED-TOC to stop the filling. Later, by detecting the symmetric event, "there is no inhibition," the ED-TOC may restart the filling. A fill-react cycling then maintains the process safely hovering near its maximum reaction rate, allowing a robust and practically time-optimal operation of the bioreactor. An experimental study case of a wastewater treatment process application is presented. There the dissolved oxygen concentration was used to detect the events needed to drive the controller.     

Reproductive biology and spatiotemporal patterns of spawning in striped marlin Kajikia audax

This study presents the first histology‐based assessment of the reproductive dynamics of south‐west Pacific striped marlin Kajikia audax. Maturity and reproductive status were assessed from histological sections of ovaries (n = 234) and testes (n = 243) of fish caught in commercial longline and recreational fisheries between 2006 and 2009. Spawning peaked in the Coral Sea during November and December at sea surface temperatures between 24·8 and 28·3° C. Lower jaw fork length (L_LJF) at 50% maturity (L_LJF50), a key variable for stock assessment, was estimated to be 2100 ± 102 mm (mean + s.e .) for females and 1668 ± 18 mm for males. Unlike large pelagic tunas Thunnus spp., the proportion of females increased with length and spawning fish formed multiple large‐scale aggregations within a broad latitudinal band. This study provides a starting point for biological parameters needed for stock assessment and conservation of K. audax and introduces the multiple aggregation spawning concept as a reproductive mechanism to explain genetic heterogeneity observed in some highly migratory species.     

Probing of C-terminal lysine variation in a recombinant monoclonal antibody production using Chinese hamster ovary cells with chemically defined media

C-terminal lysine (C-K) variants are commonly observed in therapeutic monoclonal antibodies and recombinant proteins. Heterogeneity of C-K residues is believed to result from varying degree of proteolysis by endogenous carboxypeptidase(s) during cell culture production. The achievement of batch-to-batch culture performance and product quality reproducibility is a key cell culture development criterion. Understanding the operational parameters affecting C-K levels provides valuable insight into the cell culture process. A CHO cell line X expressing a recombinant antibody was selected as the model cell line due to the exhibited sensitivity of its C-K level to the process conditions. A weak cation exchange chromatography (WCX) method with or without carboxypeptidase B (CpB) treatment was developed to monitor the C-K level for in-process samples. The effects of operating conditions (i.e., temperature and culture duration) and media trace elements (copper and zinc) on C-K variants were studied. The dominant effect on C-K level was identified as the trace elements concentration. Specifically, increased C-K levels were observed with increase of copper concentration and decrease of zinc concentration in chemically defined medium. Further, a hypothesis for C-K processing with intracellular and extracellular carboxypeptidase activity was proposed, based on preliminary intracellular carboxypeptidase Western blot results and the extracellular HCCF holding study.     

Comparative metabolite analysis to understand lactate metabolism shift in Chinese hamster ovary cell culture process

A metabolic shift from lactate production (LP) to net lactate consumption (LC) phenotype was observed in certain Chinese hamster ovary (CHO) cell lines during the implementation of a new chemically defined medium (CDM) formulation for antibody production. In addition, this metabolic shift typically leads to process performance improvements in cell growth, productivity, process robustness, and scalability. In our previous studies, a correlation between a key media component, copper, and this lactate metabolism shift was observed. To further investigate this phenomenon, two complementary studies were conducted. In the first study, a single cell line was cultivated in two media that only differed in their copper concentrations, yet were known to generate an LP or LC phenotype with that cell line. In the second study, two different cell lines, which were known to possess inherently different lactate metabolic characteristics, were cultivated in the same medium with a high level of copper; one cell line produced lactate throughout the duration of the culture, and the other consumed lactate after an initial period of LP. Cell pellet and supernatant samples from both studies were collected at regular time intervals, and their metabolite profiles were investigated. The primary finding from the metabolic analysis was that the cells in LP conditions exhibited a less efficient energy metabolism, with glucose primarily being converted into pyruvate, sorbitol, lactate, and other glycolytic intermediates. This decrease in energy efficiency may be due to an inability of pyruvate and acetyl-CoA to progress into the TCA cycle. The lack of progression into the TCA cycle or overflow metabolism in the LP phenotype resulted in the inadequate supply of ATP for the cells. As a consequence, the glycolysis pathway remained the major source of ATP, which in turn, resulted in continuous LP throughout the culture. In addition, the accumulation of free fatty acids was observed; this was thought to be a result of phospholipid catabolism that was being used to supplement the energy produced through glycolysis in order to meet the needs of LP cells. A thorough review of the metabolic profiles indicated that the lactate metabolic shift could be related to the oxidative metabolic capacity of cells.     

Comprehensive proteome analysis in Cenococcum geophilum Fr. as a tool to discover drought-related proteins

Cenococcum geophilum is a widely distributed ectomycorrhizal fungus potentially playing a significant role in resistance and resilience mechanisms of its tree hosts exposed to drought stress. In this study, we performed a large scale protein analysis in pure cultures of C. geophilum in order to gain first global insights into the proteome assembly of this fungus. Using 1-D gel electrophoresis coupled with ESI-MS/MS, we indentified 638 unique proteins. Most of these proteins were related to the metabolic and cellular processes, and the transport machinery of cells. In a second step, we examined the influence of water deprivation on the proteome of C. geophilum pure cultures at three time points of gradually imposed drought. The results indicated that 12 proteins were differentially abundant in mycelia subjected to drought compared to controls. The induced responses in C. geophilum point towards regulation of osmotic stress, maintainance of cell integrity, and counteracting increased levels of reactive oxygen species formed during water deprivation.