bluuebunny/crossref_metadata_2025 huggingface dataset:
EXTRACTION, PARTIAL PURIFICATION AND CHARACTERIZATION OF PHOSPHOENOLPYRUVATE CARBOXYKINASE FROM ASCOPHYLLIUM NODOSUM (PHAEOPHYCEAE)1
Authors: Johnston Andrew M. , Raven John A.
Published: November 2006

Phosphoenolpyruvate carboxykinase (PEPCK) from Ascophyllum nodosum (L.) Le Jolis was partially purified and characterized to investigate its role in inorganic carbon assimilation in macroalgae. Inorganic carbon isotopic disequilibrium studies showed that the carboxylation of phosphoenolpyruvate utilized CO2 rather than HCO3−as its source of inorganic carbon. This is consistent with the enzyme being a phosphoenolpyruvate carboxykinase rather than a phosphoenolpyruvate carboxylase. Pre‐incubation with Mn2+alone activated PEPCK more effectively than when combinations of Mn2+, ADP and HCO3−were used as activators. Activation of PEPCK during catalysis was found not to occur. Although the activation of PEPCK reduced the Km for CO2 by a factor of 2.25, the value reported here of 1.084 mM CO2 for the activated enzyme at pH 7.0 is at the top of the range of previously reported values for brown algal PEPCK. The specific activity of PEPCK was increased from 0.268 μmol·min−1·mg−1in the crude extract to 33.03 μmol·min−1·mg−1in the partially purified preparations. Whether PEPCK can act as an initial carboxylating enzyme is discussed.

Triton X‐100 at 0.57% (v/v) was found to be the optimum detergent and concentration for the extraction of enzymes from A. nodosum. When high concentrations of detergents ‐were used, a low (NH4)2SO4 cut was required to remove the free detergent from solution, which was extracted by centrifugation. Q Sepharose was used to partially purify PEPCK and separate it from pyruvate kinase. Good protein separations were consistently obtained.
VEGETATIVE REPRODUCTION AND SEXUAL LIFE CYCLE OF THE TOXIC DINOFLAGELLATEGYMNODINIUM CATENATUMFROM TASMANIA, AUSTRALIA1
Authors: Blackburn Susan I. , Hallegraeff Gustaaf M. , Bolch Christopher J.
Published: November 2006

The toxic, chain‐forming dinoflagellateGymnodinium catenatumGraham was cultured from vegetative cells and benthic resting cysts isolated from estuarine waters in Tasmania, Australia. Rapidly dividing, log phase cultures formed long chains of up to 64 cells whereas stationary phase cultures were composed primarily of single cells (23‐41 pm long, 27‐36 pm wide). Vegetative growth (mean doubling time 3‐4 days) was optimal at temperatures from 14.5‐20° C, salinities of 23‐34% and light irradiances of 50‐300 μE·m−2·s−1.

The sexual life cycle ofG. catenatumwas easily induced in a nutrient‐deficient medium, provided compatible opposite mating types were combined (heterothallism). Gamete fusion produced a large (59‐73 μm long, 50‐59 μm wide) biconical, posteriorly biflagellate planozygote (double longitudinal flagellum) which after several days lost one longitudinal flagellum and gradually became subspherical in shape. This older planozygote stage persisted for up to two weeks before encysting into a round, brown resting cyst (42‐52 μm diam; hypnozygote) with microreticulate surface ornamentation. Resting cysts germinated after a dormancy period as short as two weeks under our culture conditions, resulting in a single, posteriorly biflagellate germling cell (planomeiocyte). This divided to form a chain of two cells, which subsequently re‐established a vegetative population. Implications for the bloom dynamics of this toxic dinoflagellate, a causative organism of paralytic shellfish poisoning, are discussed.
Regulating MHC expression for cellular therapeutics
Authors: Figueiredo Constança , Horn Peter A. , Blasczyk Rainer , Seltsam Axel
Published: November 2006

BACKGROUND: In the past decade, regenerative medicine and cell‐based therapies have emerged as new science and technology, with the main goal of repairing, replacing, or regenerating new tissues. A critical issue in this field is the high polymorphism of HLA, which compromises immune acceptance. The lentivirus‐mediated delivery of short‐hairpin RNAs (shRNAs) has proved to be an efficient method to inhibit the translation of a specific gene.

STUDY DESIGN AND METHODS: A lentiviral‐based vector system was used for drug‐inducible expression of shRNA sequences that target either β2‐microglobulin (β2m) or HLA heavy‐chain transcripts.

RESULTS: The transduction of inducible RNA interference cassettes containing the sequences for shRNAs targeting β2m or HLA heavy chain suppressed HLA class I expression by up to 90 percent in HeLa and B‐lymphocyte cell lines as well as in peripheral blood monocytes. The expression of HLA class I antigens was fully restored in these cells after the drug had been discontinued. It was demonstrated that HLA class I knockdown was effective in preventing antibody‐mediated cell lysis and CD8+ T‐cell response. The residual HLA expression in HLA‐silenced cells may provide sufficient protection against natural killer cell–mediated lysis.

CONCLUSIONS: These data demonstrate the feasibility of controlling HLA expression by genetically modifying cell‐based therapeutics to overcome the limitations of immune rejection, bringing cellular therapies closer to reality.
Duffy and Kidd blood group antigens: minor histocompatibility antigens involved in renal allograft rejection?
Authors: Lerut Evelyne , Van Damme Boudewijn , Noizat‐Pirenne France , Emonds Marie‐Paule , Rouger Philippe , Vanrenterghem Yves , Pirenne Jacques , Ansart‐Pirenne Hélène
Published: November 2006

BACKGROUND: Minor histocompatibility antigens have been poorly defined. Whether Duffy (FY) and Kidd (JK), polymorphic and immunogenic blood group antigens, widely distributed in human organs, expressed and functional in the kidney, could function as minor histocompatibility antigens and be implicated in renal allograft rejection was questioned.

STUDY DESIGN AND METHODS: A retrospective, homogeneous, single‐center cohort of 370 renal transplants was analyzed. In all donor/recipient pairs, FY and JK polymorphisms were identified by real‐time polymerase chain reaction. In all donor/recipient pairs the matching (m) or mismatching (mm) status was defined for both systems. All biopsies were reviewed, and historical screening results for FY and JK alloantibodies and graft survival were retrospectively analyzed.

RESULTS: Although graft survival was not different between the groups, it was observed that FY mm grafts had significantly more chronic lesions compared to FY m grafts. HLA‐DR11 was more frequent in both recipients (p = 0.0081) and donors (p = 0.0104) of FY mm couples without chronic allograft nephropathy, suggesting a protective effect for this molecule. JK mm grafts had more interstitial inflammation than JK m grafts (p = 0.0369).

CONCLUSION: This renal model unmasks for the first time the role of FY and—to a lesser extent—JK antigens as minor histocompatibility antigens and suggests their potential role for other clinical transplant settings.
False‐reactive microbiologic screening test results in Swedish blood donors—how big is the problem? A survey among blood centers and deferred donors
Authors: Tynell Elsa , Norda Rut , Ekermo Bengt , Sanner Margareta , Andersson Sören , Björkman Anders
Published: November 2006

BACKGROUND: Screening of blood donors for markers of transfusion‐transmissible infectious agents leads to a varying number of false‐reactive test results and sometimes thereby temporary or permanent deferral of donors and also to loss of collected units.

STUDY DESIGN AND METHODS: Data on false‐reactive screening test results in 2002 and 2003 were collected from 19 blood centers in Sweden. A questionnaire was sent to donors deferred because of false‐reactive screening test results to investigate their perception of the information and their reaction to the deferral.

RESULTS: Testing of 21,189 samples from new donors and 423,543 donations from regular and/or repeat donors produced 1,059 false‐reactive test results, mostly from hepatitis C virus antibody testing, and 299 deferrals. Six different human immunodeficiency virus tests led to between 0.02 and 0.2 percent false‐reactive results. The deferral rate varied considerably between different counties. Of 204 deferred donors contacted, 180 (88%) answered the questionnaire. More than 80 percent were worried about their test results and worry was more common among those who did not feel sufficiently informed.

CONCLUSION: The results imply that there is a need for a more standardized approach to the screening of blood donors and donations with the aim of minimizing the number of false‐reactive screening test results. They also emphasize the importance of appropriate information and support to deferred donors.
Managing Boundaries in American Administration: The Collaboration Imperative
Authors: Kettl Donald F.
Published: November 2006

Boundaries have long played a central role in American public administration. In part, this is because boundaries are central to the administrative process, as they define what organizations are responsible for doing and what powers and functions lie elsewhere. It is also because of the nation’s political culture and unusual system of federalism, in which boundaries have always been the focus of conflict. Five boundaries have historically been important in the American administrative system: mission, resources, capacity, responsibility, and accountability. New forces make managing these boundaries increasingly difficult: political processes that complicate administrative responses, indirect administrative tactics, and wicked problems that levy enormous costs when solutions fail. Working effectively at these boundaries requires new strategies of collaboration and new skills for public managers. Failure to develop these strategies—or an instinct to approach boundaries primarily as political symbolism—worsens the performance of the administrative system.
Collaboration Processes: Inside the Black Box
Authors: Thomson Ann Marie , Perry James L.
Published: November 2006

Social science research contains a wealth of knowledge for people seeking to understand collaboration processes. The authors argue that public managers should look inside the “black box” of collaboration processes. Inside, they will find a complex construct of five variable dimensions: governance, administration, organizational autonomy, mutuality, and norms. Public managers must know these five dimensions and manage them intentionally in order to collaborate effectively.
The Design and Implementation of Cross‐Sector Collaborations: Propositions from the Literature
Authors: Bryson John M. , Crosby Barbara C. , Stone Melissa Middleton
Published: November 2006

People who want to tackle tough social problems and achieve beneficial community outcomes are beginning to understand that multiple sectors of a democratic society—business, nonprofits and philanthropies, the media, the community, and government—must collaborate to deal effectively and humanely with the challenges. This article focuses on cross‐sector collaboration that is required to remedy complex public problems. Based on an extensive review of the literature on collaboration, the article presents a propositional inventory organized around the initial conditions affecting collaboration formation, process, structural and governance components, constraints and contingencies, outcomes, and accountability issues.
Citizen‐Centered Collaborative Public Management
Authors: Cooper Terry L. , Bryer Thomas A. , Meek Jack W.
Published: November 2006

Civic engagement and collaborative public management are concepts that are defined broadly, making theoretical explication challenging and practical application of empirical research difficult. In this article, the authors adopt definitions of civic engagement and collaborative public management that are centered on the citizen and the potential for active citizenship. Following a historical review of civic engagement in the United States, a conceptual model of five approaches to civic engagement is offered. Citizen‐centered collaborative public management is enhanced through these approaches. The authors suggest the need for further empirical research on collaborative public management that is grounded in citizenship action.
The Marine Pathogenic Genotype of Spironucleus barkhanus from Farmed Salmonids Redescribed as Spironucleus salmonicida n. sp.
Authors: JØRGENSEN ANDERS , STERUD ERIK
Published: November 2006

ABSTRACT. There are two genotypes of the diplomonad Spironucleus barkhanus. Based on sequence data from the small subunit ribosomal RNA gene the conspecificity of these two genotypes has been questioned. Therefore, we have sampled Spironucleus from 27 fish, representing 14 populations, five species, and four genera. Partial nucleotide sequences from the three genes; small subunit ribosomal DNA, glutamate dehydrogenase 1 and α‐tubulin were compared. The pathogenic isolates of S. barkhanus, which causes systemic spironucleosis in Atlantic salmon, Chinook salmon, and Arctic charr, all farmed in sea water, were genetically very different from the commensal isolate found in wild freshwater populations of Arctic charr and grayling. The genetic distances between the genotypes were of the same magnitude as those separating species of Giardia. Based on these genetic and ecological data, the pathogenic genotype from farmed salmonids is described as a new species, Spironucleus salmonicida n. sp. Scanning and transmission electron microscopy showed no specific morphological or ultrastructural features distinguishing S. salmonicida n. sp. from S. barkhanus. The present study clearly demonstrates the value of applying genetics in identification of Spironucleus species. Phylogenetic analyses that included the isolates of S. salmonicida n. sp. did not change the phylogenetic relationship within the genus Spironucleus.
Implants in children with ectodermal dysplasia: a case report and literature review
Authors: Kramer Franz‐Josef , Baethge Carola , Tschernitschek Harald
Published: November 2006

Abstract: The replacement of teeth by implants is usually restricted to patients with completed craniofacial growth. Implant insertions in children or adolescents are circumvented due to several unfavorable potential effects including trauma to tooth germs, tooth eruption disorders and multidimensional restrictions of skeletal craniofacial growth. Moreover, the functional and esthetic results of the oral rehabilitation are only temporary acceptable. However, to a small number of pediatric patients suffering congenitally from severe hypodontia caused by syndromes such as ectodermal dysplasia, conventional prosthodontic rehabilitations are insufficient. We report the case of a boy with ectodermal dysplasia who exhibited a severe hypodontia and who was treated with implants inserted into the anterior mandible at the age of 8 years. The implants were functionally loaded and resulted in a high patient satisfaction. We recommend the early insertion of dental implants in children with severe hypodontia. Reviewing the current literature, several aspects of syndromic hypodontia, patient selection and implant planning are discussed.
Sensitizations to allergens of TRUE test® in 864 consecutive eczema patients in Israel
Authors: Magen Eli , Mishal Joseph , Schlesinger Menachem
Published: November 2006

The TRUE test® is a widespread diagnostic tool for initial patch testing of patients with contact dermatitis (CD). From 2002 to 2005, 864 patients with eczema were patch‐tested using TRUE test® in one Israeli allergology clinic. 547 (63.3%) patients were female and 317 (36.7%) were male. 346 (40%) patients had ≥1 positive patch test reactions. The most common allergens were nickel sulfate for 114 (13.2%) patients, potassium dichromate 111 (12.8%), fragrance mix 59 (6.8%), cobalt chloride 12 (1.4%), ethylenediamine dihydrochloride 11 (1.3%), epoxy resin 11 (1.3%), balsam of Peru 9 (1.0%), carba mix 7 (0.8%), thiomersal 6 (0.7%), wool alcohol 5 (0.6%), black rubber (PPD) mix 5 (0.6%), neomycin 4 (0.5%); Kathon CG, Colophony and Quaternium 15 – each 2 (0.2%), other allergens – each 1 (0.1%). In male patients, carba mix, black rubber (PPD) mix and epoxy resin sensitivity was more frequent, whereas nickel sulfate, fragrance mix, ethylenediamine dihydrochloride and cobalt chloride sensitivity was significantly more frequent in female patients. Our results are in general agreement with previously published reports, excluding the low sensitivity rates to cobalt, which maybe is missed by TRUE test®.
Cutaneous and systemic sequelae from exposure to 2‐vinylpyridine
Authors: Rajpar S. F. , Jackson J. R. , Foulds I. S.
Published: November 2006

We present the case of an employee of a chemical production factory who became sensitized to 2‐vinylpyridine despite wearing full protective polyvinyl chloride clothing. He developed severe dermatitis at the site of contact, secondary eczematization over the flexures and periungual areas, as well as marked systemic upset. Pyridines are known sensitizers although this reaction pattern has not previously been described.
Allergic contact dermatitis to panthenol and cocamidopropyl PG dimonium chloride phosphate in a facial hydrating lotion
Authors: Roberts Hugh , Williams Jason , Tate Bruce
Published: November 2006

Dexpanthenol is the alcohol corresponding to pantothenic acid (the water‐soluble vitamin B5). Although it is a common ingredient in many pharmaceuticals and cosmetics, contact allergy is relatively uncommon. Cocamidopropyl PG dimonium chloride phosphate is a phospholipid complex derived from pure coconut oil, and contact allergy is rare. We report a case of allergic contact dermatitis to panthenol and cocamidopropyl PG dimonium chloride phosphate in a facial hydrating lotion.
The effect of population diversity on skin irritation
Authors: Peters Lisa , Marriott Marie , Mukerji Bhashkar , Indra Pinky , Iyer Janardhana Vaidyanathan , Roy Adip , Rowson Matt , Ahmed Sharmin , Cooper Karen , Basketter David
Published: November 2006

The impact of many human variables on the response to skin irritating substances has been studied to varying degrees, including the impact of age, sex, and atopic status. However, the importance of ethnic origin has been more difficult to investigate, leading to a relative paucity of compelling data, either for or against the existence of differences. A primary reason for this lack is that studies on different ethnic groups often have to be undertaken in different locations thus introducing variables, e.g. time, environmental conditions that confound interpretations. In the present work, an attempt has been made to eliminate all variables except ethnicity by conducting a study on 2 distinct populations (Punjabis and Tamils) at the same location on the same day with a single assessor of the skin reactions, using sodium lauryl sulfate as the skin irritant. The skin reactions were assessed visually, and it was demonstrated that the modality of the reactions in these 2 populations had clear differences, but that the dose–response profiles were very similar. Thus, although the irritant response was expressed differently (e.g. erythema was much less evident in the darker Tamil population), the overall outcome was that the populations reacted in an equivalent manner.
Male‐to‐male differences in post‐thaw motility of rhesus spermatozoa after cryopreservation of replicate ejaculates
Authors: Leibo S.P. , Michael Kubisch H. , Dee Schramm R. , Harrison Richard M. , VandeVoort Catherine A.
Published: November 2006

Background The efficiency of controlled propagation to produce rhesus monkeys of particular genotypes can be maximized by use of cryopreserved spermatozoa collected from specific males to inseminate appropriate females. But this assumes that semen from males with different genotypes can be cryopreserved with equal effectiveness.

Methods To investigate whether spermatozoa from differentMacaca mulattamales can be effectively cryopreserved when frozen under identical conditions, we collected and froze semen specimens from 13 adult, fertile males maintained at three primate research centers.

Results Survival, based on post‐thaw motility normalized to the pre‐freeze value, was assayed within 30 minutes after thawing; it varied from 50% to 70% but declined thereafter. To examine the response of semen from individual males, we collected and froze three to six ejaculates per male from each of seven males.

Conclusions In general, semen from a given male responded reproducibly to freezing, but there were significant differences among males. The cause of these differences amongM. mulattamales in post‐thaw sperm survival remains unidentified.
Regulation of calprotectin expression by interleukin‐1α and transforming growth factor‐β in human gingival keratinocytes
Authors: Hayashi N. , Kido J. , Suryono  , Kido R. , Wada C. , Kataoka M. , Shinohara Y. , Nagata T.
Published: November 2006

Background and Objective:  Calprotectin, a heterodimer of S100A8 and S100A9 with antimicrobial properties, is expressed in gingival keratinocytes and plays an important role in innate immunity. Because calprotectin expression is localized in the spinous cell layer of the gingival epithelium, we hypothesized that the expression of calprotectin in keratinocytes is related to the differentiation stage. The aim of the present study was to investigate the relationship between calprotectin expression and keratinocyte differentiation using some factors that regulated its differentiation.

Material and Methods:  Normal human gingival keratinocytes were isolated from gingival tissues obtained at the extraction of wisdom teeth, and were cultured in serum‐free keratinocyte medium supplemented with interleukin‐1α or calcium, which promote keratinocyte differentiation, and transforming frowth factor‐β (TGF‐β) or retinoic acid, which suppress its differentiation. The expression of S100A8/A9 mRNA and the production of calprotectin in normal human gingival keratinocytes were examined by northern blotting and enzyme‐linked immunosorbent assay, respectively. The expression of cytokeratin 14, involucrin and filaggrin (marker proteins of keratinocyte differentiation) was investigated by immunohistochemical staining, and the DNA‐binding activity of CCAAT/enhancer binding protein α (C/EBPα), a transcription factor, was examined by electrophoretic mobility shift assay.

Results:  The expression of S100A8/A9 mRNA and the production of calprotectin were increased by interleukin‐1α and calcium, but decreased by TGF‐β. RA inhibited the expression of S100A8/A9 and keratinocyte differentiation, which were induced by interleukin‐1α. C/EBPα DNA‐binding activity in normal human gingival keratinocytes was enhanced by interleukin‐1α and calcium, but suppressed by TGF‐β.

Conclusion:  The present study suggests that calprotectin expression is related to keratinocyte differentiation and that C/EBPα is a regulator of calprotectin expression in keratinocytes.
Lip piercing: prevalence of associated gingival recession and contributing factors. A cross‐sectional study
Authors: Kapferer I. , Benesch T. , Gregoric N. , Ulm C. , Hienz S. A.
Published: November 2006

Background and Objective:  Body piercings have increased tremendously in popularity in recent years. For oral piercing, late complications include gingival trauma and localized periodontitis. The purpose of this cross‐sectional study was to investigate the prevalence and contributing factors of gingival recession associated with labial piercing.

Material and Methods:  The test group included 50 subjects with lower‐lip studs. Nonpierced controls were matched according to gender, age and smoking status. Clinical examination included plaque and bleeding indices, probing depth, recession, clinical attachment level, width of keratinized gingiva, periodontal biotype, insertion of frenula, evaluation of hard tissues, trauma of occlusion, characteristics of the stud, radiographs and photographs.

Results:  Gingival recessions were noted on teeth opposite the stud in 68% of the test group compared with 4% of the controls. Localized periodontitis was recorded in 4% of test subjects. Time since piercing and the position of the stud in relation to the cemento–enamel junction were significantly associated with the prevalence of buccal recessions. There were no significant associations between piercing and abnormal tooth wear.

Conclusion:  The prevalence of gingival recessions is associated with labial piercing. The position of the intra‐oral disc and time since piercing are associated with a greater prevalence of gingival recession. A narrow width of keratinized gingiva is associated with increased buccal recessions.
Identification and functional characterization of an Src homology domain 3 domain‐binding site on Cbl
Authors: Sanjay Archana , Miyazaki Tsuyoshi , Itzstein Cecile , Purev Enkhtsetseg , Horne William C. , Baron Roland
Published: November 2006

Cbl is an adaptor protein and ubiquitin ligase that binds and is phosphorylated by the nonreceptor tyrosine kinase Src. We previously showed that the primary interaction between Src and Cbl is mediated by the Src homology domain 3 (SH3) of Src binding to proline‐rich sequences of Cbl. The peptide Cbl RDLPPPPPPDRP(540–551), which corresponds to residues 540–551 of Cbl, inhibited the binding of a GST–Src SH3 fusion protein to Cbl, whereas RDLAPPAPPPDR(540–551) did not, suggesting that Src binds to this site on Cbl in a class I orientation. Mutating prolines 543–548 reduced Src binding to the Cbl 479–636 fragment significantly more than mutating the prolines in the PPVPPR(494–499) motif, which was previously reported to bind Src SH3. Mutating Cbl prolines 543–548 to alanines substantially reduced Src binding to Cbl, Src‐induced phosphorylation of Cbl, and the inhibition of Src kinase activity by Cbl. Expressing the mutated Cbl in osteoclasts induced a moderate reduction in bone‐resorbing activity and increased amounts of Src protein. In contrast, disabling the tyrosine kinase‐binding domain of full‐length Cbl by mutating glycine 306 to glutamic acid, and thereby preventing the previously described binding of the tyrosine kinase‐binding domain to the Src phosphotyrosine 416, had no effect on Cbl phosphorylation, the inhibition of Src activity by full‐length Cbl, or bone resorption. These data indicate that the Cbl RDLPPPP(540–546) sequence is a functionally important binding site for Src.
KEEN TO HELP? MANAGERS' IMPLICIT PERSON THEORIES AND THEIR SUBSEQUENT EMPLOYEE COACHING
Authors: HESLIN PETER A. , VANDEWALLE DON , LATHAM GARY P.
Published: November 2006

Although coaching can facilitate employee development and performance, the stark reality is that managers often differ substantially in their inclination to coach their subordinates. To address this issue, we draw from and build upon a body of social psychology research that finds that implicit person theories (IPTs) about the malleability of personal attributes (e.g., personality and ability) affect one's willingness to help others. Specifically, individuals holding an “entity theory” that human attributes are innate and unalterable are disinclined to invest in helping others to develop and improve, relative to individuals who hold the “incremental theory” that personal attributes can be developed. Three studies examined how managers' IPTs influence the extent of their employee coaching. First, a longitudinal field study found that managers' IPTs predicted employee evaluations of their subsequent employee coaching. This finding was replicated in a second field study. Third, an experimental study found that using self‐persuasion principles to induce incremental IPTs increased entity theorist managers' willingness to coach a poor performing employee, as well as the quantity and quality of their performance improvement suggestions.
Effects of successive seasons of genetically modified herbicide‐tolerant maize cropping on weeds and invertebrates
Authors: Heard M.S. , Clark S.J. , Rothery P. , Perry J.N. , Bohan D.A. , Brooks D.R. , Champion G.T. , Dewar A.M. , Hawes C. , Haughton A.J. , May M.J. , Scott R.J. , Stuart R.S. , Squire G.R. , Firbank L.G.
Published: November 2006

The use of genetically modified herbicide‐tolerant (GMHT) crops influences the abundance of weeds and some invertebrate groups because the associated herbicide regime contrasts with that of conventional systems. However, it is not clear to what extent these effects might be cumulative; should GMHT crops be grown continuously. In northern Europe, in the near future, this situation is most likely to apply to maize crops. Here, we consider the effects of continuous GMHT maize cropping on plant and invertebrate taxa using a split‐field experiment. Half of each field was managed using GMHT and the other half with a conventional variety, with the treatments retained for two seasons. The treatment effects were broadly consistent with those found in the larger sample of non‐continuous maize sites within the Farm Scale Evaluations. There was little evidence of effects being significantly more pronounced in the second year; any cumulative differences in above‐ground biodiversity between GMHT and conventional cropping were too variable to be readily detected.
Characterisation of cucumber leaf spot virus isolated from recycled irrigation water of soil‐less cucumber cultures
Authors: Rosner A. , Lachman O. , Pearlsman M. , Feigelson L. , Maslenin L. , Antignus Y.
Published: November 2006

Greenhouse‐grown cucumber plants showed mosaic‐type symptoms and irregular yellow spots on their leaves. The disease did not affect plant growth and the fruits remained symptom free. A virus having isometric particles, 30 nm in size, was isolated from the infected tissues and from recycled drainage water collected from tuff (volcanic rock) raised beds on which plants were grown. The virus was identified as a variant of cucumber leaf spot virus (CLSV) that has a host range similar but not identical to that of a previously described CLSV isolate. The overall nucleotide sequence identity between the RNAs of the Israeli isolate and the type isolate virus (accession numbers: DQ227315 and AY571334, respectively) amounts to 96%.
Long‐term outcome of therapy for 59 cats and 11 dogs with cryptococcosis
Authors: O’Brien CR , Krockenberger MB , Martin P , Wigney DI , Malik R
Published: November 2006

Objective  To determine the outcome of therapy in cats and dogs with naturally occurring cryptococcosis.

Design  Retrospective study of 59 cats and 11 dogs at the University Veterinary Centre Sydney from 1986 to 2004.

Method  Following detailed analysis of case notes potential associations between patient characteristics, cryptococcal species, retroviral status (cats), disease severity and type of therapy were examined in relation to duration and success of therapy. Treatment protocols based on amphotericin B, fluconazole and itraconazole were compared.

Results  Seventy‐six percent of feline patients were successfully treated. For cats, the presence of central nervous system disease was the only factor found to influence outcome. Cats with neurological involvement, disseminated disease or refractory disease treated with amphotericin B containing protocols did as well, on average, as cats with less severe disease treated with azole monotherapy. Amphotericin B was thus an effective agent for treating severe cases of cryptococcosis. The median cumulative dose of amphotericin B for cats cured at the first attempt was 16 mg/kg (range 7 to 23 mg/kg). The median duration of treatment required to effect a cure at first attempt was significantly shorter for fluconazole (4 months; range 1 to 8 months) than for itraconazole (9 months; range 3 to 24 months; P = 0. 0191; Mann Whitney U test). The success rate for treatment of canine cases was 55%. No factor appeared to influence disease outcome in dogs. Large cumulative doses of amphotericin B could be administered via the subcutaneous route in both species and generally with minimal nephrotoxicity. Recrudescence occurred in a significant proportion of animals, in some cases despite a reduction of serum latex cryptococcal antigen agglutination test to zero.

Conclusion  Although the prognosis of cryptococcosis should be described as guarded, a majority of the canine and especially feline patients can be expected to be cured, although treatment is protracted and expensive.
Transient renal tubulopathy in a racing Greyhound
Authors: Abraham LA , Tyrrell D , Charles JA
Published: November 2006

A 2‐year‐old female Greyhound was presented for inappetence and lethargy. On referral, results of diagnostic tests indicated renal glucosuria, increased excretion of selected amino acids and abnormal fractional excretion of electrolytes consistent with renal tubular dysfunction. Systemic blood pressure was elevated. Renal biopsy revealed mild proximal renal tubular damage consistent with a subacute toxic or hypoxic insult. Systemic hypertension, renal glucosuria and altered fractional excretion of electrolytes resolved during the 7 day period of hospital treatment. The Greyhound resumed training without recurrence of renal dysfunction.
Reference limits for urinary fractional excretion of electrolytes in adult non‐racing Greyhound dogs
Authors: Bennett SL , Abraham LA , Anderson GA , Holloway SA , Parry BW
Published: November 2006

Objective To determine reference limits for urinary fractional excretion of electrolytes in Greyhound dogs.

Methods Urinary fractional excretion was calculated using a spot clearance method preceded by a 16 to 20 hour fast in 48 Greyhound dogs. Raw data analysed using the bootstrap estimate was used to calculate the reference limits.

Results The observed range for urinary fractional excretion in Greyhound dogs was 0.0 to 0.77% for sodium, 0.9 to 14.7% for potassium, 0 to 0.66% for chloride, 0.03 to 0.22% for calcium and 0.4 to 20.1% for phosphate. Expressed as percentages, the suggested reference limits for fractional excretion in Greyhound dogs are as follows: sodium ≤ 0.72, potassium ≤ 12.2, chloride ≤ 0.55, calcium ≤ 0.13 and phosphate ≤ 16.5.

Clinical significance Veterinary practitioners may use these reference limits for urinary electrolyte fractional excretion when investigating renal tubular disease in Greyhound dogs.
Paleogenomics of Echinoderms
Authors: Bottjer David J. , Davidson Eric H. , Peterson Kevin J. , Cameron R. Andrew
Published: November 2006

Paleogenomics propels the meaning of genomic studies back through hundreds of millions of years of deep time. Now that the genome of the echinoid
Strongylocentrotus purpuratus
is sequenced, the operation of its genes can be interpreted in light of the well-understood echinoderm fossil record. Characters that first appear in Early Cambrian forms are still characteristic of echinoderms today. Key genes for one of these characters, the biomineralized tissue stereom, can be identified in the
S. purpuratus
genome and are likely to be the same genes that were involved with stereom formation in the earliest echinoderms some 520 million years ago.
Transcrystalline Melt Migration and Earth's Mantle
Authors: Schiano Pierre , Provost Ariel , Clocchiatti Roberto , Faure François
Published: November 2006

Plate tectonics and volcanism involve the formation, migration, and interaction of magma and gas. Experiments show that melt inclusions subjected to a thermal gradient migrate through olivine crystals, under the kinetic control of crystal-melt interface mechanisms. Exsolved gas bubbles remain fixed and eventually separate from the melt. Scaled to thermal gradients in Earth's mantle and geological times, our results account for the grain-scale segregation of primitive melts, reinterpret CO
2
-rich fluid inclusions as escaped from melt, and question the existence of a free, deeply percolating fluid phase. Melt migration experiments also allow us to quantify crystal growth kinetics at very low undercoolings in conditions appropriate to many natural systems.
Microtubule-Severing Activity of
Shigella
Is Pivotal for Intercellular Spreading
Authors: Yoshida Sei , Handa Yutaka , Suzuki Toshihiko , Ogawa Michinaga , Suzuki Masato , Tamai Asuka , Abe Akio , Katayama Eisaku , Sasakawa Chihiro
Published: November 2006

Some pathogenic bacteria actually invade the cytoplasm of their target host cells. Invasive bacteria acquire the propulsive force to move by recruiting actin and inducing its polymerization. Here we show that
Shigella
movement within the cytoplasm was severely hindered by microtubules and that the bacteria destroyed surrounding microtubules by secreting VirA by means of the type III secretion system. Degradation of microtubules by VirA was dependent on its α-tubulin–specific cysteine protease–like activity.
virA
mutants did not move within the host cytoplasm and failed to move into adjacent cells.
Isotopic Evidence for Dietary Variability in the Early Hominin
Paranthropus robustus
Authors: Sponheimer Matt , Passey Benjamin H. , de Ruiter Darryl J. , Guatelli-Steinberg Debbie , Cerling Thure E. , Lee-Thorp Julia A.
Published: November 2006

Traditional methods of dietary reconstruction do not allow the investigation of dietary variability within the lifetimes of individual hominins. However, laser ablation stable isotope analysis reveals that the δ
13
C values of
Paranthropus robustus
individuals often changed seasonally and interannually. These data suggest that
Paranthropus
was not a dietary specialist and that by about 1.8 million years ago, savanna-based foods such as grasses or sedges or animals eating these foods made up an important but highly variable part of its diet.
Dynamics of the Immune Response against Extracellular Products of Group A Streptococci during Infection
Authors: Maripuu Linda , Eriksson Anna , Eriksson Björn , Pauksen Karlis , Holm Stig , Norgren Mari
Published: November 2006

The immune response against the infecting group A streptococcus (GAS) extracellular products (EP) was determined in acute- and convalescent-phase sera from 75 patients with different clinical manifestations of GAS infection. All EP elicited a high proliferative response in human peripheral blood mononuclear cells. In patients with bacteremia, low neutralization in acute-phase sera was associated with development of streptococcal toxic shock syndrome. Lack of neutralization in acute-phase sera was more common in patients infected with the T1
emm1
serotype. The majority of patients did not develop the ability to neutralize the mitogenic activity of their infecting isolate despite a significant increase in enzyme-linked immunosorbent assay titer in early convalescent-phase sera. In patients with the ability to neutralize GAS EP, the immune response remained high over at least 3 years. In contrast, the neutralization capacity conferred by intravenous immunoglobulin and/or plasma treatment disappeared within 3 months.
Comparative Evaluation of Löwenstein-Jensen Proportion Method, BacT/ALERT 3D System, and Enzymatic Pyrazinamidase Assay for Pyrazinamide Susceptibility Testing of
Mycobacterium tuberculosis
Authors: Singh Pushpendra , Wesley Clement , Jadaun G. P. S. , Malonia Sunil Kumar , Das R. , Upadhyay P. , Faujdar J. , Sharma P. , Gupta P. , Mishra Abhay Kumar , Singh Kalpana , Chauhan D. S. , Sharma V. D. , Gupta U. D. , Venkatesan K. , Katoch V. M.
Published: November 2006

Pyrazinamide (PZA) is an important first-line antituberculosis drug because of its sterilizing activity against semidormant tubercle bacilli. In spite of its very high in vivo activity, its in vitro activity is not apparent unless an acidic environment is available, which makes PZA susceptibility testing difficult by conventional methods. The present study was, therefore, planned to assess the performance of the colorimetric BacT/ALERT 3D system and compare the results with those from conventional tests, i.e., the Löwenstein-Jensen (LJ) proportion method (pH 4.85) and Wayne's pyrazinamidase (PZase) assay, using 107 clinical isolates. The concordance among all of these tests was 89.71% after the first round of testing and reached 92.52% after resolution of the discordant results by retesting. Prolonged incubation of the PZase tube for up to 10 days was found to increase the specificity of the PZase test. The concordances between LJ proportion and BacT/ALERT 3D, LJ proportion and the PZase assay, and BacT/ALERT 3D and the PZase assay were found to be 99.06%, 93.46%, and 92.52%, respectively. Using the LJ results as the gold standard, the sensitivities of BacT/ALERT 3D and the PZase assay were 100 and 82.85%, respectively, while the specificity was 98.61% for both of the tests. The difference between the sensitivities of BacT/ALERT 3D and the PZase assay was significant (
P
= 0.025). The mean turnaround times for the detection of resistant and susceptible results by BacT/ALERT 3D were 8.04 and 11.32 days, respectively. While the major limitations associated with the PZase assay and the LJ proportion method are lower sensitivity in previously treated patients and a longer time requirement, respectively, the BacT/ALERT 3D system was found to be rapid, highly sensitive, and specific.
Combined Use of Pastorex Staph-Plus and Either of Two New Chromogenic Agars, MRSA ID and CHROMagar MRSA, for Detection of Methicillin-Resistant
Staphylococcus aureus
Authors: Compernolle Veerle , Verschraegen Gerda , Claeys Geert
Published: November 2006

We describe the search toward a fast and reliable strategy to detect and confirm the presence of methicillin-resistant
Staphylococcus aureus
(MRSA) in screening samples. First, we evaluated the sensitivities and specificities of oxacillin resistance screening agar (ORSA) with enrichment (tryptic soy broth [TSB] and ORSA [TSB-ORSA]) and without enrichment (ORSA), MRSA ID (MRSA_ID) plates, and CHROMagar MRSA (C_MRSA) plates, all of which were inoculated with equal volumes of a suspension made by emulsifying screening swabs. Whereas the sensitivities after 48 h were similar for all media tested (77% for MRSA_ID and ORSA; 73% for C_MRSA and ORSA after enrichment [TSB-ORSA]), the specificities of MRSA_ID (98% after 24 h and 94% after 48 h) and C_MRSA (98% after 24 h and 90% after 48 h) were superior to the specificities of ORSAs (92% after 24 h and 83% after 48 h) and TSB-ORSA (86% after 24 h and 81% after 48 h). Subsequently, the performance of the Pastorex Staph-Plus agglutination test with presumptive MRSA isolates taken directly from chromogenic agars (direct_Pastorex agglutination) was compared to that of the Pastorex Staph-Plus agglutination test with isolates from blood agar subcultures (conventional_Pastorex agglutination). When the direct_Pastorex agglutination test on MRSA_ID plates was combined with Gram staining, the direct_Pastorex agglutination test with samples from MRSA_ID plates was as reliable as the conventional_Pastorex agglutination test with samples from blood agar subcultures from MRSA_ID plates. In contrast, the direct_Pastorex agglutination test with samples from C_MRSA plates gave false-negative results. Finally, we calculated the processing times of the four different strategies, namely, (i) enrichment in TSB supplemented with NaCl, subsequent culture on ORSA, and the conventional_Pastorex agglutination test; (ii) direct inoculation of ORSA combined with conventional_Pastorex agglutination test; (iii) direct inoculation of MRSA_ID plates combined with Gram staining and the direct_Pastorex agglutination test; and (iv) direct inoculation of C_MRSA plates combined with Gram staining and the direct_Pastorex agglutination test. We concluded that the use of MRSA_ID in combination with Gram staining and the direct_Pastorex agglutination test is faster and more specific than the other strategies tested.
Penile, Urethral, and Seminal Sampling for Diagnosis of Human Papillomavirus Infection in Men
Authors: Giovannelli Lucia , Migliore Maria Colomba , Capra Giuseppina , Caleca Maria Pia , Bellavia Carmelina , Perino Antonio , Viviano Enza , Matranga Domenica , Ammatuna Pietro
Published: November 2006

Methods that used specimens from three genital sites (penile brushing [PB], urethral brushing [UB], and the retrieval of semen [SE]) from 50 men were examined for human papillomavirus (HPV) DNA detection. The rates of detection by PB, UB, SE, PB and UB, and PB and SE were 88.9%, 50.0%, 33.3%, 100%, and 97.2%, respectively. The use of PB and UB appears to be the most accurate method; as an alternative to UB, the use of SE with PB could be used to improve the rate of HPV DNA detection in men.
Quantitative Detection of Plasma Human Immunodeficiency Virus Type 2 Subtype A RNA by the Nuclisens EasyQ Assay (Version 1.1)
Authors: Rodés Berta , Sheldon Julie , Toro Carlos , Cuevas Laureano , Pérez-Pastrana Esperanza , Herrera Inmaculada , Soriano Vincent
Published: November 2006

No commercial viral load assay has yet been approved for use for measurement of human immunodeficiency virus type 2 (HIV-2) RNA levels in plasma. We assessed the performance of the NucliSens EasyQ (version 1.1) assay (EasyQ; bioMérieux, Boxtel, The Netherlands) to quantify HIV-2 viremia. A viral stock was prepared from an HIV-2 (subtype A)-infected patient. Culture supernatant was subjected to viral particle counting by electron microscopy. Serial dilutions of the viral stock were made in HIV-negative plasma and were used to test EasyQ for its sensitivity, linearity, and reproducibility. RNA was quantified by the NucliSens EasyQ (version 1.1) assay. Plasma samples from 75 HIV-2-infected patients were further tested. EasyQ was able to quantify HIV-2 RNA in a reproducible manner. Overall, estimates of the number of HIV-2 RNA copies/ml obtained with EasyQ were lower than those obtained by electron microscopy; however, the differences were always less than 0.7 log (mean, 0.55 ± 0.19 log
10
). The assay showed good linearity (
r
2
= 0.964;
P
< 0.0001). The agreement between both measures was assessed by use of a Bland-Altman plot; the narrow limits (0.158 to 0.952), defined as the mean difference ± 2 standard deviations, indicated good agreement. The reproducibility was also good, since the between-run coefficients of variation were 1.49, 3.60, and 12.25% for samples containing 6.30, 4.30, and 2.30 log
10
HIV-2 RNA copies/ml, respectively. HIV-2 RNA was detected in 34 of 75 (45%) plasma specimens (mean, 2.72 log RNA copies/ml; range, 1.74 to 4.11 log RNA copies/ml); the rest of the specimens were considered to have undetectable viremia. A negative correlation was found between the number of HIV-2 RNA copies/ml and CD4 counts. In summary, EasyQ was shown to be reliable for the measurement of plasma HIV-2 subtype A RNA levels and may be a feasible tool for routine clinical monitoring of HIV-2 subtype A-infected patients.
Characterization of In Vitro Biofilm-Associated Pneumococcal Phase Variants of a Clinically Relevant Serotype 3 Clone
Authors: McEllistrem M. Catherine , Ransford Jennifer V. , Khan Saleem A.
Published: November 2006

An increasing proportion of children with acute otitis media due to
Streptococcus pneumoniae
have serotype 3 infections since licensure of the seven-valent pneumococcal conjugate vaccine. These serotype 3 strains are genetically related by molecular subtyping. During otitis media with effusion and recurrent otitis media, biofilms commonly develop. Pneumococcal in vitro biofilms are comprised of phase variants that differ in colony morphology. By using a representative strain of the mucoid serotype 3 clone, rough phase variants with a diverse array of mutations were detected in biofilms formed in vitro. Most phase variants had mutations in the
cps3D
gene, the first gene of the capsular operon. Eleven had single nucleotide polymorphisms (SNPs) in the
cps3D
gene, one had an SNP in the −10 promoter, and three had large deletions in the
cps3D
gene. Reversion to the mucoid phenotype was associated with reversion of the mutation in the
cps3D
gene. Unlike the phase variants detected in the nasopharynx, which have at least 20% of the parental amount of capsule, the in vitro biofilm-associated phase variants had ≤12% of the parental amount of capsule, as determined by capsule enzyme-linked immunosorbent assays. Using real-time reverse transcription-PCR, we determined that capsule expression in the phase variants was likely regulated at multiple levels. These in vitro phase variation data, which underscore the plasticity of the pneumococcus, need to be confirmed with in vivo analyses of the middle ear mucosa during otitis media.