bluuebunny/crossref_metadata_2025 huggingface dataset:
Evolution of the fish rhabdovirus viral haemorrhagic septicaemia virus
Authors: Einer-Jensen Katja , Ahrens Peter , Forsberg Roald , Lorenzen Niels
Published: April 2004

Viral haemorrhagic septicaemia (VHS) caused by the rhabdovirus VHSV is economically the most important viral disease in European rainbow trout farming. Until 1989, this virus was mainly isolated from freshwater salmonids but in the last decade, it has also been isolated from an increasing number of free-living marine fish species. To study the genetic evolution of VHSV, the entire G gene from 74 isolates was analysed. VHSV from wild marine species caught in the Baltic Sea, Skagerrak, Kattegat, North Sea, and English Channel and European freshwater isolates, appeared to share a recent common ancestor. Based on the estimated nucleotide substitution rate, the ancestor of the European fresh water isolates was dated some 50 years ago. This finding fits with the initial reports in the 1950s on clinical observations of VHS in Danish freshwater rainbow trout farms. The study also indicates that European marine VHSV and the North American marine line separated approx. 500 years ago. The codon substitution rate among the freshwater VHSV isolates was found to be 2·5 times faster than among marine isolates. The data support the hypothesis of the marine environment being the original reservoir of VHSV and that the change in host range (to include rainbow trout) may have occurred several times. Virus from the marine environment will therefore continue to represent a threat to the trout aquaculture industry.
The science of the unknowable: Stafford Beer's cybernetic informatics
Authors: Pickering Andrew
Published: April 2004

This paper explores the history of Stafford Beer's work in management cybernetics, from his early conception and simulation of an adaptive automatic factory and associated experimentation in biological computing, through the development of the Viable System Model and the Team Syntegrity technique for discussion and planning. It also pursues Beer into the fields of micro‐ and macropolitics and spirituality. The aim is to show that all of Beer's projects can be understood as specific instantiations and workings out of a cybernetic ontology of unknowability and becoming: a stance that recognises that the world can always surprise us and that we can never dominate it through knowledge. The thrust of Beer's work was, thus, to construct systems that could adapt performatively to environments they could not fully control.
City planning
Authors: Schwaninger Markus , Koerner Markus
Published: April 2004

Urban development in fast‐growing cities is one of the huge challenges of our time, and several projects of technical cooperation are dedicated to this issue. The aim of this paper is to help project managers to enhance their capability of dealing effectively with the formidable complexities inherent in this kind of project. For this purpose, we explore the potential of Organizational Cybernetics and Social Systems Theory in a relatively new area of application. We have developed a set of conceptual tools that are helpful in coping with dynamic complexity in change and development projects. These tools have in common an inherent logic deriving to a great extent from Stafford Beer's Viable System Model and the St Gall framework for systemic management. The application of the tools is illustrated by a state‐of‐the‐art case study from the realm of Technical Co‐operation – the revision of the Urban Master Plan for the City of Addis Ababa, the capital of Ethiopia. However, the toolkit is in principle also applicable to any complex project of change or development.
Critical professional education about information and communications technologies and social life
Authors: Kling Rob
Published: April 2004

Looking back over the 1990s, it is easy to see the widespread troubles of many ventures that depended upon advanced IT applications, including business process re‐engineering projects, enterprise systems, knowledge management projects, online distance education courses, and, famously, some of the dot‐com businesses. These “troubles” vary from substantial underperformance (i.e. projects that were much more costly and/or produced much less social or business value than most of the participating IT professionals anticipated) and many outright failures. Many of these “troubles” could have been avoided (or at least ameliorated) if the participating IT professionals had much more reliable and critical understanding of the relationships between IT configurations, socio‐technical interventions, social behavior of other participants in different roles, and the dynamics of organizational and social change. Social informatics is the name of the field that studies and theorizes this topic, and is discussed in more detail in this paper. The key issue addressed in this paper is who will produce social informatics research for IT professionals, and where will they learn about important findings, theories, design approaches, etc.? The paper examines the record of computer science in the US as a major contributor to the relevant research and teaching. It also examines the possibilities for new kinds of academic programs – sometimes called “information schools” and “IT schools” – that are being developed to expand beyond the self‐imposed boundaries of computer science and to integrate some organizational and social research as sites for social informatics.
The customer is always right, but whose customer is more important?
Authors: Eschenfelder Kristin
Published: April 2004

This paper takes a social shaping of technology approach to identify and explain sources of conflict in the design or enhancement of corporate Web sites. Data from a multi‐case field study show how Web site classification schemes embedded in Web site design elements created intra‐organizational conflicts because the schemes could not equally accommodate different sub‐units' customer requirements. Interview data demonstrate Web managers' perceptions that Web classification schemes privileged certain sets of customer needs, and Web managers' actions to shape the design of classification schemes to satisfy their perceived customer needs. Data analysis identified three design elements of Web sites associated with sub‐unit conflict: classification categories, templates and tool bars, and database entities and attributes.
Focusing on the invisible: the representation of IS in Egypt
Authors: Shoib Gamila M. , Jones Matthew R.
Published: April 2004

Despite discussions of the increasingly global character of information systems (IS), IS research remains highly “Western‐centric” both in terms of its subjects of study and the nationalities of the authors. Researchers interested in IS in non‐Western settings are thus reliant on a fragmented and not easily accessible literature that presents a potentially distorted picture of IS practice in these regions. This paper explores this situation through an examination of the literature relating to IS in Egypt (both directly, but also as a Middle Eastern, Arab or Islamic country). A macro analysis of these studies indicates that they are predominantly positivistic in epistemology, quantitative in methodology and focused on economic development and national culture. Although many of the studies make comparisons with other countries in the region, the explicit, or sometimes implicit, point of reference is almost invariably with “the West”. This is repeated in a detailed analysis of individual studies. Implications of these findings for research on IS in non‐Western settings are discussed.
Incident management systems in disaster management
Authors: Perry Ronald W.
Published: April 2004

Particularly since the 11 September terrorist attacks in the USA, much attention has been given to the development and implementation of incident management systems (IMS). The IMS is a tool for marshalling pre‐identified and pre‐assembled resources to respond to an emergency or disaster. IMS is particularly useful when personnel and resources from many agencies and jurisdictions are required to manage large incidents successfully. While many IMS have been devised over the years, their use remains intermittent. This paper traces the evolution of IMS, reviews how it can be integrated into jurisdictional emergency and disaster management, and specifies the structures that are used in most incident management systems at the municipal level.
A cost‐benefit analysis of R&D tax incentives
Authors: Russo Benjamin
Published: April 2004

Abstract.  Although technical knowledge generates spillover benefits, production of technical knowledge creates congestion externalities; thus, private R&D investment could be inefficient. A computable general equilibrium model is used to rank tax incentives by their effects on research effort and measure welfare effects. Five results stand out: R&D tax credits produce relatively large increases in research effort and welfare. Lower corporate income tax rates and ITCs for downstream users of high‐tech production inputs rank second. Revenue losses from lower personal income tax rates can produce welfare losses. Ironically, ITCs for upstream producers of innovative inputs are ineffective. Incremental R&D credits dominate comprehensive credits. JEL Classification: E62, H21, O38
How Software Project Risk Affects Project Performance: An Investigation of the Dimensions of Risk and an Exploratory Model*
Authors: Wallace Linda , Keil Mark , Rai Arun
Published: April 2004

To reduce the high failure rate of software projects, managers need better tools to assess and manage software project risk. In order to create such tools, however, information systems researchers must first develop a better understanding of the dimensions of software project risk and how they can affect project performance. Progress in this area has been hindered by: (1) a lack of validated instruments for measuring software project risk that tap into the dimensions of risk that are seen as important by software project managers, and (2) a lack of theory to explain the linkages between various dimensions of software project risk and project performance. In this study, six dimensions of software project risk were identified and reliable and valid measures were developed for each. Guided by sociotechnical systems theory, an exploratory model was developed and tested. The results show that social subsystem risk influences technical subsystem risk, which, in turn, influences the level of project management risk, and ultimately, project performance. The implications of these findings for research and practice are discussed.
Visual and olfactory cues for mate orientation behaviour in male white‐spotted longicorn beetle, Anoplophora malasiaca
Authors: Fukaya Midori , Akino Toshiharu , Yasuda Tetsuya , Yasui Hiroe , Wakamura Sadao
Published: April 2004

Olfactory and visual cues were shown to mediate short‐distance orientation in Anoplophora malasiaca (Thomson) (Coleoptera: Cerambycidae). In a laboratory test, more than 80% (n = 42) of males walked straight upward when presented with an untreated surface with a 75° slope. When a freshly killed female was fixed at a short distance (10 cm ahead and 5 cm to left/right) from the starting point, 50% of males (n = 30) were oriented toward the female before direct contact. Similar behavioural responses were observed when female extract was directly applied to the slope or to a glass rod model fixed on the slope. When black, white, and transparent coloured rods with the extract were presented, the orientation response was significantly greater for black than to white and transparent rods, to which only a negligible response was observed.
The Fuzzy Front End of New Product Development for Discontinuous Innovations: A Theoretical Model
Authors: Reid Susan E. , De Brentani Ulrike
Published: April 2004

The fuzzy front end of the new product development (NPD) process, the time and activity prior to an organization's first screen of a new product idea, is the root of success for firms involved with discontinuous new product innovation. Yet understanding the fuzzy front‐end process has been a challenge for academics and organizations alike. While approaches to handling the fuzzy front end have been suggested in the literature, these tend to be relevant largely for incremental new product situations where organizations are aware of and are involved in the NPD process from the project's beginning. For incremental new products, structured problems or opportunities typically are laid out at the organizational level and are directed to individuals for information gathering. In the case of discontinuous innovations, however, we propose that the process works in the opposite direction—that is, that the timing and likelihood of organizational‐level involvement is more likely to be at the discretion of individuals. Such individuals perform a boundary‐spanning function by identifying and by understanding emerging patterns in the environment, with little or no direction from the organization. Often, these same individuals also act as gatekeepers by deciding on the value to the organization of externally derived information, as well as whether such information will be shared. Consequently for discontinuous innovations, information search and related problems/opportunities are unstructured and are at the individual level during the fuzzy front end. As such, the direction of initial decisions about new environmental information tends to be inward, toward the corporate decision‐making level, rather than the other way around.

In order to cope with the special and complex nature of decisions made at the fuzzy front end of NPD for discontinuous innovations, this process is detailed as a series of decisions occurring over three proposed interfaces: boundary, gatekeeping, and project. The difference between each interface lies in the nature of the decisions made: At the boundary and gatekeeping interfaces, the primary impetus is individual‐level decision‐making; at the project interface, decisions occur at the organizational level. By articulating these processes in the form of a model, we achieve two objectives: (1) We outline a more detailed and comprehensive approach to understanding the nature of the front‐end decision making process for discontinuous innovations; and (2) we detail specific propositions for future research on each stage of the process.
Relevance of hyperglycemia to early mechanical hyperalgesia in streptozotocin‐induced diabetes
Authors: Romanovsky Dmitry , Hastings Stephanie L. , Stimers Joseph R. , Dobretsov Maxim
Published: April 2004

Abstract A modified von Frey filament test and an algesiometer paw pressure test were used to measure mechanical nociceptive withdrawal thresholds of the hind limb of control rats and rats injected with streptozotocin (STZ, 50 mg/kg). STZ treatment induced hyperglycemia (HG rats) in about 40% of treated animals. The rest of the STZ‐treated and control rats remained normoglycemic (NG rats) throughout the entire experiment. No indications of mechanical hyperalgesia were observed in control groups of animals injected with physiological buffer only. However, both the behavioral tests used detected a 15–30% decrease in the mechanical nociceptive threshold of rats treated with STZ. Furthermore, mechanical nociceptive threshold changes were statistically indistinguishable between NG and HG rats. Glucose tolerance test did not reveal abnormalities of glucose metabolism in NG rats (compared to control animals). However, 1 week after STZ injection, the serum insulin level of NG rats was significantly lower than that of age‐matched control rats (0.81 ± 0.16 vs. 3.5 ± 0.4 ng/mL; p < 0.01). These data strongly argue that systemic hyperglycemia is not the only factor triggering the development of mechanical hyperalgesia in the STZ rat model of diabetes. Other than hyperglycemia, consequences of insulinemia or insulinemia itself may play an important role in early impairment of mechanical nociception in this animal model.
Evaluation of topiramate as an anti‐hyperalgesic and neuroprotective agent in the peripheral nervous system
Authors: Bischofs Stefan , Zelenka Marek , Sommer Claudia
Published: April 2004

Abstract  Topiramate (TPM), a novel anti‐convulsant currently approved for the treatment of epileptic disorders, has been shown to possess neuroprotective effects in models of cerebral ischemia, status epilepticus, and facial nerve lesion. Furthermore, pilot studies showed an effect of TPM in neuropathic pain models. Here, we studied the anti‐hyperalgesic and neuroprotective efficacy of TPM in rat models of peripheral nerve lesions. Rats with a unilateral chronic constrictive injury (CCI) or a crush lesion of the sciatic nerve were treated with a twice‐daily dose of 20 mg/kg of TPM. Behavioral and neurophysiological tests were used to measure pain‐related behavior, motor, and sensory function. Morphometry was performed to evaluate sciatic nerves. In CCI, treatment with TPM attenuated mechanical hyperalgesia and cold allodynia. In sciatic nerve crush, TPM reduced cold allodynia and attenuated thermal hyperalgesia at the early and late phase of the observation. There was no difference in the numbers of surviving or regenerating nerve fibers between saline‐ and TPM‐treated rats in either model. Electrophysiological studies carried out over a period of 3 months after sciatic nerve crush did not show major differences between TPM‐ and saline‐treated rats. In conclusion, we could show moderate anti‐hyperalgesic effects but could not prove a neuroprotective effect of TPM in these two rat nerve injury models using electrophysiological and morphometric methods.
Unusual neurophysiological and immunological findings in myasthenia gravis: a case report
Authors: Scaioli Vidmer , Andreetta Francesca , Mantegazza Renato
Published: April 2004

Abstract  We describe the case of a male patient with ocular myasthenia gravis who developed a diabetic neuropathy similar to chronic inflammatory demyelinating polyradiculoneuropathy associated with transient generalized ‘myokymic’ discharges and distal weakness. He had antibodies against acetylcholine receptor and GQ1b ganglioside, but not anti‐voltage‐gated K+ channel antibodies. Serial electrophysiological and immunological findings showed that diabetes was involved in the immune‐mediated mechanism of peripheral neuropathy. We hypothesize that the concomitant appearance of distal motor weakness and decreased compound muscle action potentials upon repetitive nerve stimulation, together with increased distal motor latency and generalized peripheral nerve hyperexcitability, were all related to transient serum positivity to anti‐GQ1b antibodies.
Adaptation of the brucellae to their intracellular niche
Authors: Roop R. Martin , Bellaire Bryan H. , Valderas Michelle Wright , Cardelli James A.
Published: April 2004

Members of the bacterial genus Brucella are facultative intracellular pathogens that reside predominantly within membrane‐bound compartments within two host cell types, macrophages and placental trophoblasts. Within macrophages, the brucellae route themselves to an intracellular compartment that is favourable for survival and replication, and they also appear to be well‐adapted from a physiological standpoint to withstand the environmental conditions encountered during prolonged residence in this intracellular niche. Much less is known about the interactions of the Brucella with placental trophoblasts, but experimental evidence suggests that these bacteria use an iron acquisition system to support extensive intracellular replication within these host cells that is not required for survival and replication in host macrophages. Thus, it appears that the brucellae rely upon the products of distinct subsets of genes to adapt successfully to the environmental conditions encountered within the two cell types within which they reside in their mammalian hosts.
Staphylococcal NreB: an O2‐sensing histidine protein kinase with an O2‐labile iron–sulphur cluster of the FNR type
Authors: Kamps Annegret , Achebach Stephanie , Fedtke Iris , Unden Gottfried , Götz Friedrich
Published: April 2004

The nreABC (nitrogen regulation) operon encodes a new staphylococcal two‐component regulatory system that controls dissimilatory nitrate/nitrite reduction in response to oxygen. Unlike other two‐component sensors NreB is a cytosolic protein with four N‐terminal cysteine residues. It was shown that both the NreB–cysteine cluster and Fe ions are required for function. Isolated NreB was converted to the active form by incubation with cysteine desulphurase, ferrous ions and cysteine. This activation is typical for FeS‐containing proteins and was reversed by oxygen. During reconstitution an absorption band at 420 nm and a yellow‐brownish colour (typical for an FNR‐type iron–sulphur cluster formation) developed. After alkylation of thiol groups in NreB and in the cysteine mutant NreB(C62S) almost no iron–sulphur cluster was incorporated; both findings corroborated the importance of the cysteine residues. Comparison of the kinase activity of (i) the reconstituted (ii) the unreconstituted, and (iii) the unreconstituted and deferrated NreB–His indicated that NreB kinase activity depended on iron availability and was greatly enhanced by reconstitution. NreB is the first direct oxygen‐sensing protein described in staphylococci so far. Reconstituted NreB contains 4–8 acid‐labile Fe and sulphide ions per NreB which is in agreement with the presence of 1–2 iron–sulphur [4Fe‐4S]2+ clusters of the FNR‐type. Unlike FNR, NreB does not act directly as transcriptional activator, but transfers the phosphoryl group to the response regulator NreC.
Use of arthropods for the evaluation of the olive‐orchard management regimes
Authors: Ruano Francisca , Lozano Carlos , Garcia Pedro , Peña Aranzazu , Tinaut Alberto , Pascual Felipe , Campos Mercedes
Published: April 2004

Abstract  1 The presence and abundance of arthropods were compared in three olive orchards under organic, integrated and conventional management regimes. In each olive orchard, trees were sampled in the canopy by beating branches and soil arthropods by placing pitfall traps. Contrary to expectations, the highest abundance of arthropods occurred in the integrated management orchard. The most abundant groups were Formicidae and the species Euphyllura olivinae (Homoptera: Psyllidae).

2 Canopies and the soil under the tree canopy (interior soil) were selected as the most informative sites for sampling. The months with the strongest differences were May, June and July, especially June. In the canopy, Araneae, Coleoptera, Diptera, Heteroptera, Hymenoptera, Homoptera, Lepidoptera, Neuroptera and Thysanoptera were the most abundant, and showed significant differences in abundance among orchards with different management regimes. Moreover, in the canopies, Coleoptera and Lepidoptera showed a seasonal pattern of abundance and consistent significant differences between the organic orchard vs. the integrated and conventional ones in both years of study. In the soil, 12 orders showed significant differences in abundance among management regimes at some point of the sampling season.

3 In a search for biological indicators that could help to distinguish between management regimes, a discriminant analysis applied to the data indicated that only the samples from the canopy were classified according to their management regime in a consistent way over time. The groups selected by the analysis to establish differences among management regimes were Coleoptera, Diptera, Heteroptera, Lepidoptera and Thysanoptera. The analysis applied to compare organic vs. non‐organic olive orchards, again identified Coleoptera and Lepidoptera as suitable groups. The results suggest that these two orders are potential bioindicators to distinguish, in a simple way, organic olive orchards from non‐organic ones.
Dispersal patterns and chromatic response of Scaphoideus titanus Ball (Homoptera Cicadellidae), vector of the phytoplasma agent of grapevine flavescence dorée
Authors: Lessio Federico , Alma Alberto
Published: April 2004

Abstract  1 Scaphoideus titanus Ball, a nearctic leafhopper introduced into Europe in the 1950s, is known to be the vector of the phytoplasma agent of flavescence dorée (FD), a persistent disease of grapevine. Knowledge of its dispersal patterns is thus very important to prevent disease outbreaks.

2 Yellow sticky traps were used to study the seasonal flight activity of S. titanus, its vertical flight, its movement outside the vineyard and the influence of plant density. Sticky traps of different colours (yellow, red, blue, and white) were also compared. The behaviour of males and females was tested for all those conditions.

3 Abundance was greater in normal than in low plant density conditions, and a positive relationship was found between number of plants per square metre and presence of S. titanus. Leafhoppers did not appear capable of spreading significantly outside a vineyard. Few individuals were trapped above the canopy. Red sticky traps caught more individuals than white, yellow or blue, with the latter showing a poor attractiveness. Sex ratio was almost always male biased.

4 Scaphoideus titanus is monophagous and appears incapable of great dispersal away from its host plant, and females are less likely to fly than males. Further studies on the influence of different factors on the behaviour of this leafhopper are suggested.
Expression of an 11 kDa methionine‐rich delta‐zein in transgenic soybean results in the formation of two types of novel protein bodies in transitional cells situated between the vascular tissue and storage parenchyma cells
Authors: Kim Won‐Seok , Krishnan Hari B.
Published: April 2004

Soybean (Glycine max (L.) Merr.) is an important protein source in human diets and animal feeds. The sulphur content of soybean seed proteins, however, is not optimal for ration formulations. Thus, increasing the methionine and cysteine content of soybean seed proteins would enhance the nutritional quality of this widely utilized legume. We have earlier reported the isolation of an 11 kDa δ‐zein protein rich in methionine from the endosperm of the maize (Zea mays L.) inbred line W23a1 [Kim, W.‐S. and Krishnan, H.B. (2003) Allelic variation and differential expression of methionine‐rich‐δ‐zeins in maize inbred lines B73 and W23a1. Planta, 217, 66–74]. Using Agrobacterium‐mediated transformation, a construct consisting of the coding region of the cloned δ‐zein gene under regulation of the β‐conglycinin α′‐promoter was introduced into the soybean genome. The 11 kDa δ‐zein gene was expressed in the seeds of transgenic soybeans, although low‐level expression was also detected in the leaves. In situ hybridization indicated that the 11 kDa δ‐zein mRNA was expressed predominantly in transitional cells located between the vascular tissue and storage parenchyma cells. Immunohistochemistry of developing transgenic soybeans revealed that the accumulation of the 11 kDa δ‐zein occurred primarily in these transitional cells. Expression of the 11 kDa δ‐zein gene in transgenic soybean resulted in the formation of two endoplasmic reticulum‐derived protein bodies that were designated as either spherical or complex. Immunocytochemical localization demonstrated that both the spherical and complex protein bodies accumulated the 11 kDa δ‐zein. Although expression of the 11 kDa δ‐zein gene elevated the methionine content of the alcohol‐soluble protein fraction 1.5–1.7‐fold above that of the non‐transgenic line, the overall methionine content of seed flour was not increased. Our results suggest that the confined expression of the 11 kDa δ‐zein gene in transitional cells could be limiting the increase in methionine content in transgenic soybean seeds.
Reduction of starch granule size by expression of an engineered tandem starch‐binding domain in potato plants
Authors: Ji Qin , Oomen Ronald J. F. J. , Vincken Jean‐Paul , Bolam David N. , Gilbert Harry J. , Suurs Luc C. J. M. , Visser Richard G. F.
Published: April 2004

Granule size is an important parameter when using starch in industrial applications. An artificial tandem repeat of a family 20 starch‐binding domain (SBD2) was engineered by two copies of the SBD derived from Bacillus circulans cyclodextrin glycosyltransferase via the Pro‐Thr‐rich linker peptide from Xyn10A from Cellulomonas fimi. SBD2 and a single SBD were introduced into the amylose‐free potato mutant, amf, using appropriate signal sequences. The accumulation of SBD2 into transgenic starch granules was much higher than that of SBD. In a number of transformants, particularly amfSS3, the starch granules were much smaller than in control plants. The amfSS3 mean granule size was 7.8 µm, compared with 15.2 µm in the control, whereas other starch properties were unaltered. This new starch combines the advantage of the high purity of potato starch with that of the small granule size of other crop species, such as cassava, taro and wheat. This starch may find application in the manufacture of biodegradable plastic films. Both genes were also expressed in Escherichia coli and the affinity for soluble starch of the purified recombinant proteins was determined. SBD2 had an approximately 10‐fold higher affinity for starch than SBD, indicating that the two appended SBDs act in synergy when binding to their target polysaccharide ligand.
Accumulation of rotavirus VP6 protein in chloroplasts of transplastomic tobacco is limited by protein stability
Authors: Birch‐Machin Ian , Newell Christine A. , Hibberd Julian M. , Gray John C.
Published: April 2004

Rotavirus VP6 is a highly immunogenic major capsid protein that may be useful as a subunit vaccine. The expression of a bovine group A rotavirus VP6 cDNA was examined in tobacco chloroplasts following particle bombardment. Constructs containing the VP6 cDNA under the control of plastid rrn or psbA promoters, or the Escherichia coli trc promoter, were inserted, together with the aadA selectable marker gene, between the rbcL and accD genes of the tobacco plastid genome. The 40‐kDa VP6 protein accumulated to about 3% of total soluble protein in seedlings and young leaves of homoplasmic transplastomic plants containing the VP6 cDNA under the control of the rrn promoter. Lower amounts of VP6 (∼0.6% total soluble protein) accumulated in plants containing the VP6 cDNA under the control of the psbA promoter, and VP6 was undetectable in plants containing the VP6 cDNA under the control of the trc promoter. The VP6 protein in chloroplasts was shown to form trimers, as found in the rotavirus virion. However, the amount of VP6 protein declined as the leaves matured, although VP6 transcripts were still present, suggesting that the protein was susceptible to proteolytic degradation in chloroplasts.
Timing, Duration, and Transitions of the Last Interglacial Asian Monsoon
Authors: Yuan Daoxian , Cheng Hai , Edwards R. Lawrence , Dykoski Carolyn A. , Kelly Megan J. , Zhang Meiliang , Qing Jiaming , Lin Yushi , Wang Yongjin , Wu Jiangyin , Dorale Jeffery A. , An Zhisheng , Cai Yanjun
Published: April 2004

Thorium-230 ages and oxygen isotope ratios of stalagmites from Dongge Cave, China, characterize the Asian Monsoon and low-latitude precipitation over the past 160,000 years. Numerous abrupt changes in
18
O/
16
O values result from changes in tropical and subtropical precipitation driven by insolation and millennial-scale circulation shifts. The Last Interglacial Monsoon lasted 9.7 ± 1.1 thousand years, beginning with an abrupt (less than 200 years) drop in
18
O/
16
O values 129.3 ± 0.9 thousand years ago and ending with an abrupt (less than 300 years) rise in
18
O/
16
O values 119.6 ± 0.6 thousand years ago. The start coincides with insolation rise and measures of full interglacial conditions, indicating that insolation triggered the final rise to full interglacial conditions.
Curcumin, a Major Constituent of Turmeric, Corrects Cystic Fibrosis Defects
Authors: Egan Marie E. , Pearson Marilyn , Weiner Scott A. , Rajendran Vanathy , Rubin Daniel , Glöckner-Pagel Judith , Canny Susan , Du Kai , Lukacs Gergely L. , Caplan Michael J.
Published: April 2004

Cystic fibrosis is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR). The most common mutation, ΔF508, results in the production of a misfolded CFTR protein that is retained in the endoplasmic reticulum and targeted for degradation. Curcumin is a nontoxic Ca–adenosine triphosphatase pump inhibitor that can be administered to humans safely. Oral administration of curcumin to homozygous ΔF508 CFTR mice in doses comparable, on a weight-per-weight basis, to those well tolerated by humans corrected these animals' characteristic nasal potential difference defect. These effects were not observed in mice homozygous for a complete knockout of the CFTR gene. Curcumin also induced the functional appearance of ΔF508 CFTR protein in the plasma membranes of transfected baby hamster kidney cells. Thus, curcumin treatment may be able to correct defects associated with the homozygous expression of ΔF508 CFTR.
Regeneration of Peroxiredoxins by p53-Regulated Sestrins, Homologs of Bacterial AhpD
Authors: Budanov Andrei V. , Sablina Anna A. , Feinstein Elena , Koonin Eugene V. , Chumakov Peter M.
Published: April 2004

Acting as a signal, hydrogen peroxide circumvents antioxidant defense by overoxidizing peroxiredoxins (Prxs), the enzymes that metabolize peroxides. We show that sestrins, a family of proteins whose expression is modulated by p53, are required for regeneration of Prxs containing Cys-SO
2
H, thus reestablishing the antioxidant firewall. Sestrins contain a predicted redox-active domain homologous to AhpD, the enzyme catalyzing the reduction of a bacterial Prx, AhpC. Purified Hi95 (sestrin 2) protein supports adenosine triphosphate–dependent reduction of overoxidized PrxI in vitro, indicating that unlike AhpD, which is a disulfide reductase, sestrins are cysteine sulfinyl reductases. As modulators of peroxide signaling and antioxidant defense, sestrins constitute potential therapeutic targets.
Efficacy of Novel Hemagglutinin-Neuraminidase Inhibitors BCX 2798 and BCX 2855 against Human Parainfluenza Viruses In Vitro and In Vivo
Authors: Alymova Irina V. , Taylor Garry , Takimoto Toru , Lin Tsu-Hsing , Chand Pooran , Babu Y. Sudhakara , Li Chenghong , Xiong Xiaoping , Portner Allen
Published: April 2004

Human parainfluenza viruses are important respiratory tract pathogens, especially of children. However, no vaccines or specific therapies for infections caused by these viruses are currently available. In the present study we characterized the efficacy of the novel parainfluenza virus inhibitors BCX 2798 and BCX 2855, which were designed based on the three-dimensional structure of the hemagglutinin-neuraminidase (HN) protein. The compounds were highly effective in inhibiting hemagglutinin (HA) and neuraminidase (NA) activities and the growth of hPIV-1, hPIV-2, and hPIV-3 in LLC-MK
2
cells. The concentrations required to reduce the activity to 50% of that of a control ranged from 0.1 to 6.0 μM in HA inhibition assays and from 0.02 to 20 μM in NA inhibition assays. The concentrations required to inhibit virus replication to 50% of the level of the control ranged from 0.7 to 11.5 μM. BCX 2798 and BCX 2855 were inactive against influenza virus HA and NA and bacterial NA. In mice infected with a recombinant Sendai virus whose HN gene was replaced with that of hPIV-1 [rSV(hHN)], intranasal administration of BCX 2798 (10 mg/kg per day) and of BCX 2855 (50 mg/kg per day) 4 h before the start of infection resulted in a significant reduction in titers of virus in the lungs and protection from death. Treatment beginning 24 h after the start of infection did not prevent death. Together, our results indicate that BCX 2798 and BCX 2855 are effective inhibitors of parainfluenza virus HN and may limit parainfluenza virus infections in humans.
Molecular Mechanisms of Anti-Inflammatory Action of Erythromycin in Human Bronchial Epithelial Cells: Possible Role in the Signaling Pathway That Regulates Nuclear Factor-κB Activation
Authors: Desaki Masashi , Okazaki Hitoshi , Sunazuka Toshiaki , Omura Satoshi , Yamamoto Kazuhiko , Takizawa Hajime
Published: April 2004

Long-term macrolide therapy has been proven to improve survival in patients with diffuse panbronchiolitis. Although its mechanisms remain unknown, previous studies have suggested the effects of macrolide might be anti-inflammatory rather than antibacterial. To elucidate the molecular mechanisms of its action, we studied here the effects of erythromycin (EM) and its new derivative, EM703, which shows no antibacterial action, on the activation of the transcription factor nuclear factor-κB (NF-κB) in human bronchial epithelial cells. Western blotting analysis showed that EM did not inhibit the degradation of IκBα, suggesting the molecular target for EM was not the dissociation of NF-κB from IκB. An electrophoretic mobility shift assay showed that EM did not interrupt the NF-κB DNA-binding activity in the nucleus under the conditions tested. Moreover, not only EM but also EM703 suppressed the activation of NF-κB and the production of interleukin-8, demonstrating that the anti-inflammatory action of the macrolide is independent of its antibacterial activity. Taken together, these data suggest EM has an anti-inflammatory action, presumably via an interaction with the NF-κB signaling pathway in the downstream of the dissociation from IκB, resulting in the inhibition of NF-κB.
Genetic and Biochemical Characterization of a Chromosome-Encoded Carbapenem-Hydrolyzing Ambler Class D β-Lactamase fromShewanella algae
Authors: Héritier Claire , Poirel Laurent , Nordmann Patrice
Published: April 2004

A chromosome-encoded β-lactamase gene fromShewanella algaeclinical isolate KB-1 was cloned and expressed inEscherichia coli. It encoded the Ambler class D enzyme OXA-55, sharing less than 55% identity with any other oxacillinases. Although conferring a narrow-spectrum β-lactam resistance phenotype, OXA-55 had carbapenem-hydrolyzing activity that mirrored the reduced susceptibility to imipenem observed inS. algaeKB-1. Very similar oxacillinases were found in otherS. algaeisolates.
Role of the Multidrug Efflux System MexXY in the Emergence of Moderate Resistance to Aminoglycosides among
Pseudomonas aeruginosa
Isolates from Patients with Cystic Fibrosis
Authors: Vogne Christelle , Aires Julio Ramos , Bailly Christiane , Hocquet Didier , Plésiat Patrick
Published: April 2004

This study investigates the role of active efflux system MexXY in the emergence of aminoglycoside (AG) resistance among cystic fibrosis (CF) isolates of
Pseudomonas aeruginosa
. Three genotypically related susceptible and resistant (S/R) bacterial pairs and three other AG-resistant CF strains were compared to four non-CF strains moderately resistant to AGs. As demonstrated by immunoblot experiments, pump MexY was strongly overproduced in all of the resistant bacteria. This MexXY upregulation was associated with a 2- to 16-fold increase in the MICs of AGs in the S/R pairs and lower intracellular accumulation of dihydrostreptomycin. Alterations in
mexZ
, the repressor gene of operon
mexXY
, were found in all of the AG-resistant CF isolates and in one non-CF strain. Complementation of these bacteria with a plasmid-borne
mexZ
gene dramatically reduced the MICs of AGs, thus highlighting the role played by MexXY in the development of moderate resistance in CF patients. In contrast, complementation of the three non-CF strains showing wild-type
mexZ
genes left residual levels of resistance to AGs. These data indicate that a locus different from
mexZ
may be involved in overproduction of MexXY and that other nonenzymatic mechanisms contribute to AG resistance in
P. aeruginosa
.
Pseudomonas aeruginosa
LasA Protease in Treatment of Experimental Staphylococcal Keratitis
Authors: Barequet Irina S. , Ben Simon Guy J. , Safrin Mary , Ohman Dennis E. , Kessler Efrat
Published: April 2004

LasA protease is a staphylolytic endopeptidase secreted by
Pseudomonas aeruginosa
. We have examined the effectiveness of LasA protease in the treatment of staphylococcal keratitis caused by methicillin-sensitive
Staphylococcus aureus
(MSSA) and methicillin-resistant
S. aureus
(MRSA) isolates in a rabbit model. Keratitis was induced by intrastromal injection of the bacteria. The eyes were treated topically, and the efficacy of LasA protease was compared to those of lysostaphin (a staphylolytic protease secreted by
Staphylococcus simulans
) and vancomycin. When treatment was initiated early (4 h) after infection, practically all of the MSSA- and MRSA-infected corneas were sterilized by LasA protease, and its efficacy in eradicating the bacteria was comparable to those of lysostaphin and vancomycin. By contrast, most of the control corneas were heavily infected, with median values of 4.5 × 10
6
(MSSA) and 5 × 10
5
(MRSA) CFU/cornea (
P
< 0.001). When treatment was initiated late (10 h) after infection, LasA protease reduced the numbers of CFU in both MSSA- and MRSA-infected corneas by 3 to 4 orders of magnitude compared to the numbers of CFU for the controls (median values, 1,380 and 30 CFU/cornea, respectively, for the treated animals compared to 1.2 × 10
6
and 5 × 10
5
CFU/cornea for the respective controls [
P
= 0.001]), and it was more effective than vancomycin in eradicating MRSA cells (
P
= 0.02). In both the early- and the late-treatment protocols, the clinical scores for eyes treated with LasA protease were significantly lower than those for the eyes of the corresponding controls and comparable to those for the lysostaphin- and vancomycin-treated eyes. We conclude that LasA protease is effective in the treatment of experimental
S. aureus
keratitis in rabbits and may have potential for the treatment of disease in humans.
Mutant Selection Window in Levofloxacin and Moxifloxacin Treatments of Experimental Pneumococcal Pneumonia in a Rabbit Model of Human Therapy
Authors: Croisier Delphine , Etienne Manuel , Bergoin Emilie , Charles Pierre-Emmanuel , Lequeu Catherine , Piroth Lionel , Portier Henri , Chavanet Pascal
Published: April 2004

For some pneumococci the fluoroquinolone MICs are low but the mutant prevention concentrations (MPCs) are high; this difference defines in vitro the mutant selection window (MSW). We investigated in vivo the bacterial reduction and the occurrence of resistant mutants with moxifloxacin (MFX; 400 mg once daily) or levofloxacin (LVX; 500 mg twice daily) in treatments similar to those in humans with experimental pneumonia due to pneumococci (expPP) exhibiting various MICs and MPCs. The MIC/MPC for MFX and LVX and genotypes were as follows: strain 16089, 0.125/0.125 and 0.5/0.5 (wild type); strain MS1A, 0.25/0.25 and 1/2 (efflux); strain MS2A, 0.25/4 and 1.75/28 (
parC79
); strain MR3B4, 0.25/4 and 2/32 (
parC79
); strain M16, 0.5/2 and 8/32 (
parC83
); strain Gyr-1207, 1.5/3 and 8/16 (
gyrA
); and strain MQ3A, 4/4 and 16/64 (
parC
and
gyrA
). Both drugs were efficient with wild type-expPP, but only MFX was efficient with efflux-expPP. No bacterial reduction was observed for
parC
-expPPs due to mutants observed in 18 to 100% of animals, depending on the strain and the drug tested. These mutants showed unbound area under the concentration-time curve and MICs of from 50 to 164 for MFX. The in vivo pharmacodynamic boundaries of the MSW were different for MFX and LVX. We conclude that, after LVX or MFX treatment, mutants occur in vivo if there is a preexisting
parC
mutation, since the drug concentrations fall below the MPCs of these strains. Since the MPC determination cannot be routinely determined, these phenotypes or genotypes should be detected by simple tests to guide the therapeutic options.
Dose-Dependent Pharmacokinetics of Itraconazole after Intravenous or Oral Administration to Rats: Intestinal First-Pass Effect
Authors: Shin Jee H. , Choi Ka Y. , Kim Yu C. , Lee Myung G.
Published: April 2004

The dose-dependent pharmacokinetics of itraconazole after intravenous (10, 20, or 30 mg/kg) and oral (10, 30, or 50 mg/kg) administration and the first-pass effects of itraconazole after intravenous, intraportal, intragastric, and intraduodenal administration at a dose of 10 mg/kg were evaluated in rats. After intravenous administration at a dose of 30 mg/kg, the area under the plasma concentration-time curve from time zero to infinity (AUC0-∞) was significantly greater than those at 10 and 20 mg/kg (1,090, 1,270, and 1,760 μg · min/ml for 10, 20, and 30 mg/kg, dose-normalized at 10 mg/kg). After oral administration, the AUC0-∞was significantly different for three oral doses (380, 687, and 934 μg · min/ml for 10, 30, and 50 mg/kg, respectively, dose-normalized at 10 mg/kg). The extent of absolute oral bioavailability (F) was 34.9% after an oral dose at 10 mg/kg. The AUC0-∞(or AUC0-8 h) values were comparable between intravenous and intraportal administration and between intragastric and intraduodenal administration, suggesting that the hepatic and gastric first-pass effects were almost negligible in rats. However, the AUC0-8 hvalues after intraduodenal and intragastric administration were significantly smaller than that after intraportal administration, approximately 30%, suggesting that the intestinal first-pass effect was approximately 70% of that of an oral dose of 10 mg/kg. The lowFafter oral administration of itraconazole at a dose of 10 mg/kg could be mainly due to the considerable intestinal first-pass effect.
In Vitro Activity of Expanded-Spectrum Pyridazinyl Oxime Ethers Related to Pirodavir: Novel Capsid-Binding Inhibitors with Potent Antipicornavirus Activity
Authors: Barnard D. L. , Hubbard V. D. , Smee D. F. , Sidwell R. W. , Watson K. G. W. , Tucker S. P. T. , Reece P. A. R.
Published: April 2004

Picornaviruses (PV) include human rhinovirus (HRV), the primary cause of the common cold, and the enteroviruses (EV), which cause serious diseases such as poliomyelitis, meningoencephalitis, and systemic neonatal disease. Although no compounds for PV infections have been approved in the United States, pirodavir was one of the most promising capsid-binding compounds to show efficacy in human clinical trials for chemoprophylaxis of the common cold. Susceptibility to hydrolysis precluded its use as an oral agent. We have developed orally bioavailable pyridazinyl oxime ethers that are as potent as pirodavir. Compounds BTA39 and BTA188 inhibited a total of 56 HRV laboratory strains and three clinical isolates as determined by neutral red uptake assay. At concentrations of <100 nM, BTA39 inhibited 69% of the HRV serotypes and isolates evaluated, BTA188 inhibited 75%, and pirodavir inhibited 59% of the serotypes and isolates. The 50% inhibitory concentrations (IC
50
s) for the two compounds ranged from 0.5 nM to 6,701 nM. The compounds also inhibited EV, including coxsackie A and B viruses (IC
50
= 773 to 3,608 nM) and echoviruses (IC
50
= 193 to 5,155 nM). BTA39 only inhibited poliovirus strain WM-1 at 204 nM, and BTA188 only inhibited poliovirus strain Chat at 82 nM. EV 71 was inhibited by BTA39 and BTA188, with IC
50
s of 1 and 82 nM, respectively. Both compounds were relatively nontoxic in actively growing cells (50% cytotoxic doses, ≥4,588 nM). These data suggest that these oxime ethers warrant further investigation as potential agents for treating selected PV infections.