Construction and characterization of a live, attenuated esrB mutant of Edwardsiella tarda and its potential as a vaccine against the haemorrhagic septicaemia in turbot, Scophthamus maximus (L.)

The esrB gene of Edwardsiella tarda, which encodes a regulator protein of the type III secretion system, was mutated by the unmarked deletion method and reintroduced by allelic exchange into the chromosome of E. tarda LSE40 by means of the suicide vector pRE 112. The LSE40 esrB mutant was highly attenuated when inoculated intraperitoneally into turbot Scophthamus maximus L., showing a 50% lethal dose of 10(8.1) cfu/fish. The esrB mutants were not recoverable from the internal organs at 14 days post-inoculation. Vaccination with a single dose of 10(5)-10(7) cfu/fish of the esrB mutant elicited significant protection against the wildtype strain of E. tarda LSE40 (relative percentage survival > 50%). The protection correlated well with the antibody titres in the serum of vaccinated fish. (c) 2006 Elsevier Ltd. All rights reserved.

The Apertospathulidae, a new family of haptorid ciliates (Protozoa, Ciliophora)

We studied the morphology of three rare haptorid ciliates, using live observation and silver impregnation: Apertospathula verruculifera n. sp., Longispatha elegans n. gen., n. sp., and Rhinothrix porculus (Penard, 1922) n. gen., n. comb. Simple ethanol fixation (50-70%, v/v) is recommended to reveal the ciliary pattern of "difficult" ciliates, such as R. porculus, by protargol impregnation. The three genera investigated have a distinct feature in common, viz., a lasso-shaped oral bulge and circumoral kinety, where the right half is slightly to distinctly longer than the left and the circumoral kinety is open ventrally. Thus, they are united in a new spathidiid family, the Apertospathulidae n. fam., which probably evolved from a Bryophyllum-like ancestor by partial reduction of the oral bulge and circumoral kinety. Apertospathula verruculifera has a wart-like process, the palpus dorsalis, at the anterior end of the dorsal brush. The right branch of the circumoral kinety is only slightly longer than the left one. Longispatha elegans has a straight oral bulge and circumoral kinety, the right branch of which extends to the posterior end of the body while the left branch ends in the anterior third of the body. Rhinothrix porculus, a curious ciliate with a snout-like dorsal elongation of the oral bulge, the palpus oralis, has a highly characteristic ciliary pattern: the oral pattern is as in Longispatha, but the bulge and circumoral kinety extend spirally to the posterior end of the body while the somatic kinetics course meridionally. This is achieved by inserting some shortened kinetics in the curves of the oral bulge.

Live-weight gain, apparent digestibility, and economic benefits of yaks fed different diets during winter on the Tibetan plateau

Our goal was to determine the effect of diets with different crude protein (CP) contents and metabolizable energy (W) levels on daily live-weight gain, apparent digestibility, and economic benefit of feedlot yaks on the Tibetan plateau during winter. Yaks were either 2- or 3-years old and randomly selected from the same herd. The 3-year-olds were placed into one of two experimental groups (A and B) and a control (CK1), and the two-year-olds were placed into one of three experimental groups (C, D and E) and a control (CK2) (N per group = 5). Yak in the control groups were allow graze freely, while those in the experimental groups yaks were fed diets higher in contains crude protein and metabolizable energy through a winter period inside a warming shed. Results indicated that live-weight gain of treatment groups was higher than their respective controls during experiment, and that daily live-weight gain of every 10 days among different treatments was significant difference (P < 0.05). In addition, apparent digestibility of different diets was linearly and positively related to feedlotting time, and feed conversion efficiency for A, C, D and E groups was quadratically related to feedlotting time (P < 0.01), however, feed conversion efficiency for B group was linearly and positively related to feedlotting time (P < 0.05). The economic benefit was 1.15 for A, 1.89 for B, 1.16 for C, 1.54 for D, and 4,52 for E. (c) 2005 Elsevier B.V. All rights reserved.

A Hierarchical Characterization of a Live Streaming Media Workload

We present what we believe to be the first thorough characterization of live streaming media content delivered over the Internet. Our characterization of over five million requests spanning a 28-day period is done at three increasingly granular levels, corresponding to clients, sessions, and transfers. Our findings support two important conclusions. First, we show that the nature of interactions between users and objects is fundamentally different for live versus stored objects. Access to stored objects is user driven, whereas access to live objects is object driven. This reversal of active/passive roles of users and objects leads to interesting dualities. For instance, our analysis underscores a Zipf-like profile for user interest in a given object, which is to be contrasted to the classic Zipf-like popularity of objects for a given user. Also, our analysis reveals that transfer lengths are highly variable and that this variability is due to the stickiness of clients to a particular live object, as opposed to structural (size) properties of objects. Second, based on observations we make, we conjecture that the particular characteristics of live media access workloads are likely to be highly dependent on the nature of the live content being accessed. In our study, this dependence is clear from the strong temporal correlations we observed in the traces, which we attribute to the synchronizing impact of live content on access characteristics. Based on our analyses, we present a model for live media workload generation that incorporates many of our findings, and which we implement in GISMO [19].

Confidence intervals for half-life deviations from purchasing power parity

Existing point estimates of half-life deviations from purchasing power parity (PPP), around 3-5 years, suggest that the speed of convergence is extremely slow. This article assesses the degree of uncertainty around these point estimates by using local-to-unity asymptotic theory to construct confidence intervals that are robust to high persistence in small samples. The empirical evidence suggests that the lower bound of the confidence interval is between four and eight quarters for most currencies, which is not inconsistent with traditional price-stickiness explanations. However, the upper bounds are infinity for all currencies, so we cannot provide conclusive evidence in favor of PPP either. © 2005 American Statistical Association.

Dental microwear in live, wild-trapped Alouatta palliata from Costa Rica.

One problem with dental microwear analyses of museum material is that investigators can never be sure of the diets of the animals in question. An obvious solution to this problem is to work with live animals. Recent work with laboratory primates has shown that high resolution dental impressions can be obtained from live animals. The purpose of this study was to use similar methods to begin to document rates and patterns of dental microwear for primates in the wild. Thirty-three Alouatta palliata were captured during the wet season at Hacienda La Pacifica near Canas, Costa Rica. Dental impressions were taken and epoxy casts of the teeth were prepared using the methods of Teaford and Oyen (1989a). Scanning electron micrographs were taken of the left mandibular second molars at magnifications of 200x and 500x. Lower magnification images were used to calculate rates of wear, and higher magnification images were used to measure the size and shape of microwear features. Results indicate that, while basic patterns of dental microwear are similar in museum samples and samples of live, wild-trapped animals of the same species, ecological differences between collection locales may lead to significant intraspecific differences in dental microwear. More importantly, rates of microwear provide the first direct evidence of differences in molar use between monkeys and humans.

CIRQUE DE LA VOIX: VOCAL PERFORMANCES FOR THE 21st CENTURY AUDIENCE

This dissertation addresses the growing need to entice people to attend a classical solo vocal recital by incorporating thematic programming, multi-media presentations, collaborations and innovative marketing. It comprises four programs that use the above tactics, creating live performances of classical vocal music that appeal to the attention deficient 21st-century audience. Each program focuses on repertoire appropriate for the male alto voice and includes elements of spoken word, visual imagery and for movement through collaborations with actors, singers, dancers, designers and visual artists. Program one (March 1, 2004), La Voix Humaine: The Life of an Englishwoman in Music, Poetry, & Art, outlines the life of a fictitious Englishwoman through a self-composed narration, spoken by an actress, a Power Point presentation of visual art by 20th-century English artists and musical commentary provided by the collaboration of a vocalist and a pianist. Program two (October 15, 2004), La Voix Thfrmatique: Anima - Music that Moves, is a program of pieces ranging from the 14th- to the 20th-centuries of which half are choreographed by members of the University of Maryland Dance Department. Program three is a lecture recital entitled L 'Haute Voix: Identifying the High Male Voice and Appropriate Repertoire which is presented in collaboration with three singers, a pianist, a harpsichordist and a cellist. Program four, La Voix Dramatique: Opera Roles for the Countertenor Voice, comprises performances of George Frederic Handel's Giulio Cesare in Egitto (1724) in collaboration with the Maryland Opera Studio and the Clarice Smith Performing Arts Center (Leon Major, director; Kenneth Merrill, conductor). There are two performances each of the title role, Cesare (April 15 & 17, 2005), and his nemesis, Tolomeo (April 21 & 23,2005). All programs are documented in a digital audio format available on compact disc and are accompanied by program notes also available in digital format. Programs two and four are also documented in digital video format available on digital video disc.

A GLOBAL SAMPLING OF PIANO MUSIC FROM 1978 TO 2005

Pianists of the twenty-first century have a wealth of repertoire at their fingertips. They busily study music from the different periods -- Baroque, Classical, Romantic, and some of the twentieth century -- trying to understand the culture and performance practice of the time and the stylistic traits of each composer so they can communicate their music effectively. Unfortunately, this leaves little time to notice the composers who are writing music today. Whether this neglect proceeds from lack of time or lack of curiosity, I feel we should be connected to music that was written in our own lifetime, when we already understand the culture and have knowledge of the different styles that preceded us. Therefore, in an attempt to promote today’s composers, I have selected piano music written during my lifetime, to show that contemporary music is effective and worthwhile and deserves as much attention as the music that preceded it. This dissertation showcases piano music composed from 1978 to 2005. A point of departure in selecting the pieces for this recording project is to represent the major genres in the piano repertoire in order to show a variety of styles, moods, lengths, and difficulties. Therefore, from these recordings, there is enough variety to successfully program a complete contemporary recital from the selected works, and there is enough variety to meet the demands of pianists with different skill levels and recital programming needs. Since we live in an increasingly global society, music from all parts of the world is included to offer a fair representation of music being composed everywhere. Half of the music in this project comes from the United States. The other half comes from Australia, Japan, Russia, and Argentina. The composers represented in these recordings are: Lowell Liebermann, Richard Danielpour, Frederic Rzewski, Judith Lang Zaimont, Samuel Adler, Carl Vine, Nikolai Kapustin, Akira Miyoshi and Osvaldo Golijov. With the exception of one piano concerto, all the works are for solo piano. This recording project dissertation consists of two 60 minute CDs of selected repertoire, accompanied by a substantial document of in-depth program notes. The recordings are documented on compact discs that are housed within the University of Maryland Library System.

Magnetic resonance imaging of graded skeletal muscle injury in live rats.

INTRODUCTION: Increasing number of stretch-shortening contractions (SSCs) results in increased muscle injury. METHODS: Fischer Hybrid rats were acutely exposed to an increasing number of SSCs in vivo using a custom-designed dynamometer. Magnetic resonance imaging (MRI) imaging was conducted 72 hours after exposure when rats were infused with Prohance and imaged using a 7T rodent MRI system (GE Epic 12.0). Images were acquired in the transverse plane with typically 60 total slices acquired covering the entire length of the hind legs. Rats were euthanized after MRI, the lower limbs removed, and tibialis anterior muscles were prepared for histology and quantified stereology. RESULTS: Stereological analyses showed myofiber degeneration, and cellular infiltrates significantly increased following 70 and 150 SSC exposure compared to controls. MRI images revealed that the percent affected area significantly increased with exposure in all SSC groups in a graded fashion. Signal intensity also significantly increased with increasing SSC repetitions. DISCUSSION: These results suggest that contrast-enhanced MRI has the sensitivity to differentiate specific degrees of skeletal muscle strain injury, and imaging data are specifically representative of cellular histopathology quantified via stereological analyses.

The Macrophage-Specific Promoter mfap4 Allows Live, Long-Term Analysis of Macrophage Behavior during Mycobacterial Infection in Zebrafish.

Transgenic labeling of innate immune cell lineages within the larval zebrafish allows for real-time, in vivo analyses of microbial pathogenesis within a vertebrate host. To date, labeling of zebrafish macrophages has been relatively limited, with the most specific expression coming from the mpeg1 promoter. However, mpeg1 transcription at both endogenous and transgenic loci becomes attenuated in the presence of intracellular pathogens, including Salmonella typhimurium and Mycobacterium marinum. Here, we describe mfap4 as a macrophage-specific promoter capable of producing transgenic lines in which transgene expression within larval macrophages remains stable throughout several days of infection. Additionally, we have developed a novel macrophage-specific Cre transgenic line under the control of mfap4, enabling macrophage-specific expression using existing floxed transgenic lines. These tools enrich the repertoire of transgenic lines and promoters available for studying zebrafish macrophage dynamics during infection and inflammation and add flexibility to the design of future macrophage-specific transgenic lines.

Live Imaging of Host-Parasite Interactions in a Zebrafish Infection Model Reveals Cryptococcal Determinants of Virulence and Central Nervous System Invasion.

UNLABELLED: The human fungal pathogen Cryptococcus neoformans is capable of infecting a broad range of hosts, from invertebrates like amoebas and nematodes to standard vertebrate models such as mice and rabbits. Here we have taken advantage of a zebrafish model to investigate host-pathogen interactions of Cryptococcus with the zebrafish innate immune system, which shares a highly conserved framework with that of mammals. Through live-imaging observations and genetic knockdown, we establish that macrophages are the primary immune cells responsible for responding to and containing acute cryptococcal infections. By interrogating survival and cryptococcal burden following infection with a panel of Cryptococcus mutants, we find that virulence factors initially identified as important in causing disease in mice are also necessary for pathogenesis in zebrafish larvae. Live imaging of the cranial blood vessels of infected larvae reveals that C. neoformans is able to penetrate the zebrafish brain following intravenous infection. By studying a C. neoformans FNX1 gene mutant, we find that blood-brain barrier invasion is dependent on a known cryptococcal invasion-promoting pathway previously identified in a murine model of central nervous system invasion. The zebrafish-C. neoformans platform provides a visually and genetically accessible vertebrate model system for cryptococcal pathogenesis with many of the advantages of small invertebrates. This model is well suited for higher-throughput screening of mutants, mechanistic dissection of cryptococcal pathogenesis in live animals, and use in the evaluation of therapeutic agents. IMPORTANCE: Cryptococcus neoformans is an important opportunistic pathogen that is estimated to be responsible for more than 600,000 deaths worldwide annually. Existing mammalian models of cryptococcal pathogenesis are costly, and the analysis of important pathogenic processes such as meningitis is laborious and remains a challenge to visualize. Conversely, although invertebrate models of cryptococcal infection allow high-throughput assays, they fail to replicate the anatomical complexity found in vertebrates and, specifically, cryptococcal stages of disease. Here we have utilized larval zebrafish as a platform that overcomes many of these limitations. We demonstrate that the pathogenesis of C. neoformans infection in zebrafish involves factors identical to those in mammalian and invertebrate infections. We then utilize the live-imaging capacity of zebrafish larvae to follow the progression of cryptococcal infection in real time and establish a relevant model of the critical central nervous system infection phase of disease in a nonmammalian model.

Mapping 2-D defects in a conductive half-space by eigenfunction expansions in K-space of Fourier-Laplace transforms

info:eu-repo/semantics/published

Study of the thermal stress in a Pb-free half-bump solder joint under current stressing

The thermal stress in a Sn3.5Ag1Cu half-bump solder joint under a 3.82×108 A/m2 current stressing was analyzed using a coupled-field simulation. Substantial thermal stress accumulated around the Al-to-solder interface, especially in the Ni+(Ni,Cu)3Sn4 layer, where a maximal stress of 138 MPa was identified. The stress gradient in the Ni layer was about 1.67×1013 Pa/m, resulting in a stress migration force of 1.82×10-16 N, which is comparable to the electromigration force, 2.82×10-16 N. Dissolution of the Ni+(Ni,Cu)3Sn4 layer, void formation with cracks at the anode side, and extrusions at the cathode side were observed

Spank performance tour [at National Review of Live Art 2008]

Spank is a duet about two women’s journeys as they play out different roles. One woman recalls stinging anecdotes marked by intimacy, jealousy, fear and fantasy from her childhood in the 70s suburbs – round the corner from where Mike Leigh would film Life Is Sweet. Her Double (projected on screen) reads from a newspaper about an event that punctured the local community, creating an enduring cultural scar, her voice lacking emotion. Both women must eventually face one another and ask, who is deceiving whom? The consequences of guilt and shame and the unstable shift from private memory to public consequence is inevitable and deadly.[ABSTRACT BY THE AUTHOR]