bluuebunny/crossref_metadata_2025 huggingface dataset:
An evaluation of the Enhanced Thinking Skills (ETS) programme with mentally disordered offenders in a high security hospital
Authors: Tapp James , Fellowes Emma , Wallis Nicola , Blud Linda , Moore Estelle
Published: August 2008

Purpose. Further to evidence of the successful application of cognitive skills programmes with offenders in HM prison service, the Enhanced Thinking Skills (ETS) training course is now delivered in various secure forensic hospitals, and its usefulness with an offender patient population merits examination. Our aim was to evaluate the impact of ETS using measures designed to capture change in key areas targeted by the course, and a more global measure of mental well‐being.

Methods. Offender patients (N = 83) referred for ETS between 2001 and 2006 via a wider groupwork service within a high security hospital were administered questionnaires before and after the training course, which was minimally adjusted to meet the needs of the patient population. Attendance rates, including ‘drop‐out’ (completion of 10 or less sessions) were recorded. Demographics by subgroup (completers vs. non‐completers), and the clinical significance of any changes endorsed by participants post‐group were examined.

Results. ETS completers were significantly more likely than non‐completers to endorse changes in thinking style (e.g. a reduction in post‐course scores). Significant differences were observed at both the group and individual level in the direction of less externalization of blame, a slight increase in tolerance of frustration, an increase in social conformity, and improvement in critical reasoning skills. Furthermore there was a reduction in the endorsement of aggressive solutions to social problems.

Conclusions. These findings demonstrate a significant short‐term impact of the course for mentally disordered offenders with regards change in aspects of their thinking style and enhancement of their social problem solving skills. This profile lends some support to the provision of ETS within secure hospital settings.
Psychiatric morbidity and treatment pathway outcomes of patients presenting to specialist NHS psychodynamic psychotherapy services: Results from a multi‐centre study
Authors: Chiesa Marco. , Fonagy Peter. , Bateman Anthony W. , Mace Chris.
Published: August 2008

Objective. Little is known about socio‐demographic, diagnostic, and clinical characteristics of patients referred for assessment to psychodynamic psychotherapy services. The aim of this study was to remedy this by prospectively collecting comprehensive and systematic demographic and clinical information on a large number of patients referred to NHS psychodynamic psychotherapy services.

Design. Fourteen psychotherapy services operating within a National Health Service joined the study and contributed data for 1,136 patients referred from primary and secondary care clinics.

Method. Patients were assessed using questionnaires and self‐rated measures, which included the clinician‐based version of the diagnostic form of the Millon clinical multi‐axial inventory‐III‐revised edition (MCMI‐III‐R), the brief symptom inventory (BSI), the inventory of interpersonal problems (IIP), and the clinical outcome in routine evaluation (CORE). The pathway from assessment through to treatment and variables associated with treatment uptake and exclusion are described and examined.

Results. Most patients were in the moderate to severe range of psychiatric severity at the time of presentation. Ninety‐five percent met clinically based criteria for a psychiatric disorder (mostly anxiety and mood disorders) and/or personality disorder. Although the majority of patients were found suitable for treatment (N=935, 82%), analysis of uptake showed relatively high rates of treatment rejection by patients and treatment drop‐out. Partial outcome data at 6‐month follow‐up after intake into treatment revealed significant change but modest effect size (d=0.35).

Conclusion. Systematic collection of baseline and outcome data would provide a national database of the performance of psychotherapy services that would be invaluable in facilitating comparative studies.
Achieving Superior Financial Performance in China: Differentiation, Cost Leadership, or Both?
Authors: Li Caroline Bingxin , Li Julie Juan
Published: August 2008

Firms operating in emerging economies must make appropriate marketing strategy choices to address the idiosyncratic challenges of rapid changes and institutional voids. The authors investigate when various types of marketing strategies work by examining the effects of both pure and dual strategies on financial performance for domestic versus foreign firms and across different market concentration levels in China's emerging economy. According to a survey of 249 firms, the effectiveness of specific strategy choices varies depending on domesticity and the level of market concentration. In particular, the impacts of both cost and dual strategies on financial performance are stronger for foreign firms than for domestic firms. Although cost-leadership and dual strategies are less effective in less concentrated markets than in more concentrated ones, the effect of a differentiation strategy is stronger when the level of market concentration is low rather than high.
Intra- and Interfirm Coordination of Export Manufacturers: A Cluster Analysis of Indigenous Chinese Exporters
Authors: Zhang Chun , Hu Zuohao , Gu Flora F.
Published: August 2008

Export manufacturers face challenges associated with intrafirm coordination among functional units and interfirm coordination with foreign distributors. The majority of research on export coordination examines intra- and interfirm coordination strategies separately. Consequently, limited insights have been gained into how export manufacturers combine the various intra- and interfirm coordination strategies to achieve export success. Using a cluster analysis, the authors incorporate both intra- and interfirm coordination strategies as inputs to identify profiles of export manufacturers. They identify four distinct profiles: balanced coordinators, interactive coordinators, flexibility-only coordinators, and unfocused coordinators. The authors find that the best performers are balanced coordinators, or the export manufacturers that concurrently deploy multiple intra- and interfirm coordination strategies.
Product Complexity and Cultural Distance Effects on Managing International Distributor Relationships: A Contingency Approach
Authors: Solberg Carl Arthur
Published: August 2008

How should an exporter manage its relationship with its intermediaries given different levels of performance ambiguity? The author addresses this question by exploring four different settings of performance ambiguity that involve cultural distance (external uncertainty) and product complexity (internal uncertainty). Using a sample of 173 Norwegian exporting companies, the author analyzes relational drivers’ effect on relationship quality between trading partners—namely, exporters and their distributors. The results suggest that cultural distance and product complexity are two key contingency factors that moderate the role of relational mechanisms used by exporters in creating relationship quality with their foreign sales intermediaries. The author concludes by discussing implications for research and management.
Quantitative Trait Loci Mapping of Genome Regions Controlling Permethrin Resistance in the Mosquito Aedes aegypti
Authors: Saavedra-Rodriguez Karla , Strode Clare , Flores Suarez Adriana , Fernandez Salas Ildefonso , Ranson Hilary , Hemingway Janet , Black William C
Published: August 2008

The mosquito Aedes aegypti is the principal vector of dengue and yellow fever flaviviruses. Permethrin is an insecticide used to suppress Ae. aegypti adult populations but metabolic and target site resistance to pyrethroids has evolved in many locations worldwide. Quantitative trait loci (QTL) controlling permethrin survival in Ae. aegypti were mapped in an F3 advanced intercross line. Parents came from a collection of mosquitoes from Isla Mujeres, México, that had been selected for permethrin resistance for two generations and a reference permethrin-susceptible strain originally from New Orleans. Following a 1-hr permethrin exposure, 439 F3 adult mosquitoes were phenotyped as knockdown resistant, knocked down/recovered, or dead. For QTL mapping, single nucleotide polymorphisms (SNPs) were identified at 22 loci with potential antixenobiotic activity including genes encoding cytochrome P450s (CYP), esterases (EST), or glutathione transferases (GST) and at 12 previously mapped loci. Seven antixenobiotic genes mapped to chromosome I, six to chromosome II, and nine to chromosome III. Two QTL of major effect were detected on chromosome III. One corresponds with a SNP previously associated with permethrin resistance in the para sodium channel gene and the second with the CCEunk7o esterase marker. Additional QTL but of relatively minor effect were also found. These included two sex-linked QTL on chromosome I affecting knockdown and recovery and a QTL affecting survival and recovery. On chromosome II, one QTL affecting survival and a second affecting recovery were detected. The patterns confirm that mutations in the para gene cause target-site insensitivity and are the major source of permethrin resistance but that other genes dispersed throughout the genome contribute to recovery and survival of mosquitoes following permethrin exposure.
The Rate and Character of Spontaneous Mutation in Thermus thermophilus
Authors: Mackwan Reena R , Carver Geraldine T , Kissling Grace E , Drake John W , Grogan Dennis W
Published: August 2008

Selection of spontaneous, loss-of-function mutations at two chromosomal loci (pyrF and pyrE) enabled the first molecular-level analysis of replication fidelity in the extremely thermophilic bacterium Thermus thermophilus. Two different methods yielded similar mutation rates, and mutational spectra determined by sequencing of independent mutants revealed a variety of replication errors distributed throughout the target genes. The genomic mutation rate estimated from these targets, 0.00097 ± 0.00052 per replication, was lower than corresponding estimates from mesophilic microorganisms, primarily because of a low rate of base substitution. However, both the rate and spectrum of spontaneous mutations in T. thermophilus resembled those of the thermoacidophilic archaeon Sulfolobus acidocaldarius, despite important molecular differences between these two thermophiles and their genomes.
Rtf1-Mediated Eukaryotic Site-Specific Replication Termination
Authors: Eydmann T , Sommariva E , Inagawa T , Mian S , Klar A J S , Dalgaard J Z
Published: August 2008

The molecular mechanisms mediating eukaryotic replication termination and pausing remain largely unknown. Here we present the molecular characterization of Rtf1 that mediates site-specific replication termination at the polar Schizosaccharomyces pombe barrier RTS1. We show that Rtf1 possesses two chimeric myb/SANT domains: one is able to interact with the repeated motifs encoded by the RTS1 element as well as the elements enhancer region, while the other shows only a weak DNA binding activity. In addition we show that the C-terminal tail of Rtf1 mediates self-interaction, and deletion of this tail has a dominant phenotype. Finally, we identify a point mutation in Rtf1 domain I that converts the RTS1 element into a replication barrier of the opposite polarity. Together our data establish that multiple protein DNA and protein–protein interactions between Rtf1 molecules and both the repeated motifs and the enhancer region of RTS1 are required for site-specific termination at the RTS1 element.
A “FLP-Out” System for Controlled Gene Expression in Caenorhabditis elegans
Authors: Voutev Roumen , Hubbard E Jane Albert
Published: August 2008

We present a two-part system for conditional FLP-out of FRT-flanked sequences in Caenorhabditis elegans to control gene activity in a spatially and/or temporally regulated manner. Using reporters, we assess the system for efficacy and demonstrate its use as a cell lineage marking tool. In addition, we construct and test a dominant-negative form of hlh-12, a gene that encodes a basic helix-loop-helix (bHLH) transcription factor required for proper distal tip cell (DTC) migration. We show that this allele can be conditionally expressed from a heat-inducible FLP recombinase and can interfere with DTC migration. Using the same DTC assay, we conditionally express an hlh-12 RNAi-hairpin and induce the DTC migration defect. Finally, we introduce a set of traditional and Gateway-compatible vectors to facilitate construction of plasmids for this technology using any promoter, reporter, and gene/hairpin of interest.
HP1 Is Distributed Within Distinct Chromatin Domains at Drosophila Telomeres
Authors: Frydrychova Radmila Capkova , Mason James M , Archer Trevor K
Published: August 2008

Telomeric regions in Drosophila are composed of three subdomains. A chromosome cap distinguishes the chromosome end from a DNA double-strand break; an array of retrotransposons, HeT-A, TART, and TAHRE (HTT), maintains telomere length by targeted transposition to chromosome ends; and telomere-associated sequence (TAS), which consists of a mosaic of complex repeated sequences, has been identified as a source of gene silencing. Heterochromatin protein 1 (HP1) and HP1-ORC-associated protein (HOAP) are major protein components of the telomere cap in Drosophila and are required for telomere stability. Besides the chromosome cap, HP1 is also localized along the HTT array and in TAS. Mutants for Su(var)205, the gene encoding HP1, have decreased the HP1 level in the HTT array and increased transcription of individual HeT-A elements. This suggests that HP1 levels directly affect HeT-A activity along the HTT array, although they have little or no effect on transcription of a white reporter gene in the HTT. Chromatin immunoprecipitation to identify other heterochromatic proteins indicates that TAS and the HTT array may be distinct from either heterochromatin or euchromatin.
The multiple-wing-hairs Gene Encodes a Novel GBD–FH3 Domain-Containing Protein That Functions Both Prior to and After Wing Hair Initiation
Authors: Yan Jie , Huen David , Morely Terri , Johnson Glynnis , Gubb David , Roote John , Adler Paul N
Published: August 2008

The frizzled signaling/signal transduction pathway controls planar cell polarity (PCP) in both vertebrates and invertebrates. Epistasis experiments argue that in the Drosophila epidermis multiple wing hairs (mwh) acts as a downstream component of the pathway. The PCP proteins accumulate asymmetrically in pupal wing cells where they are thought to form distinct protein complexes. One is located on the distal side of wing cells and a second on the proximal side. This asymmetric protein accumulation is thought to lead to the activation of the cytoskeleton on the distal side, which in turn leads to each cell forming a single distally pointing hair. We identified mwh as CG13913, which encodes a novel G protein binding domain–formin homology 3 (GBD–FH3) domain protein. The Mwh protein accumulated on the proximal side of wing cells prior to hair formation. Unlike planar polarity proteins such as Frizzled or Inturned, Mwh also accumulated in growing hairs. This suggested that mwh had two temporally separate functions in wing development. Evidence for these two functions also came from temperature-shift experiments with a temperature-sensitive allele. Overexpression of Mwh inhibited hair initiation, thus Mwh acts as a negative regulator of the cytoskeleton. Our data argued early proximal Mwh accumulation restricts hair initiation to the distal side of wing cells and the later hair accumulation of Mwh prevents the formation of ectopic secondary hairs. This later function appears to be a feedback mechanism that limits cytoskeleton activation to ensure a single hair is formed.
Increase in Quantitative Variation After Exposure to Environmental Stresses and/or Introduction of a Major Mutation: G × E Interaction and Epistasis or Canalization?
Authors: Zhang Xu-Sheng
Published: August 2008

Why does phenotypic variation increase upon exposure of the population to environmental stresses or introduction of a major mutation? It has usually been interpreted as evidence of canalization (or robustness) of the wild-type genotype; but an alternative population genetic theory has been suggested by J. Hermisson and G. Wagner: “the release of hidden genetic variation is a generic property of models with epistasis or genotype–environment interaction.” In this note we expand their model to include a pleiotropic fitness effect and a direct effect on residual variance of mutant alleles. We show that both the genetic and environmental variances increase after the genetic or environmental change, but these increases could be very limited if there is strong pleiotropic selection. On the basis of more realistic selection models, our analysis lends further support to the genetic theory of Hermisson and Wagner as an interpretation of hidden variance.
Free Radical Injury and Blood-Brain Barrier Permeability in Hypoxic-Ischemic Encephalopathy
Authors: Kumar Ashok , Mittal Roopali , Khanna Hari Dev , Basu Sriparna
Published: August 2008

OBJECTIVES. The purpose of this work was to evaluate the extent of free radical injury in newborns with hypoxic ischemic encephalopathy by measuring plasma levels of malondialdehyde and nitric oxide and to assess the blood-brain barrier permeability by measuring the cerebrospinal fluid albumin/plasma albumin ratio.

METHODS. This prospective observational study was conducted over a period of 2 years at Sir Sundarlal Hospital, Banaras Hindu University. The study population consisted of 43 term neonates with perinatal asphyxia who subsequently developed hypoxic ischemic encephalopathy. Twenty normal gestational age- and gender-matched healthy infants without any perinatal asphyxia served as control subjects. Peripheral venous blood samples were analyzed for malondialdehyde, total plasma nitrates/nitrites, and albumin levels between 12 and 24 hours of life. To assess the blood-brain barrier permeability, the cerebrospinal fluid albumin/plasma albumin ratio was measured. Correlation among the levels of malondialdehyde, nitrates/nitrites, and blood-brain barrier permeability was calculated. Data were analyzed by using SPSS 10 software.

RESULTS. Plasma malondialdehyde and nitrate/nitrite levels were significantly higher in infants with hypoxic ischemic encephalopathy compared with control subjects. Although there was a progressive increment in plasma levels of malondialdehyde with increasing severity of hypoxic ischemic encephalopathy, the differences were not statistically significant. Plasma nitrate/nitrite levels were almost similar in all stages of hypoxic ischemic encephalopathy. Plasma albumin levels were comparable in infants with hypoxic ischemic encephalopathy and control subjects, whereas cerebrospinal fluid albumin levels and blood-brain barrier permeability were significantly higher in infants with hypoxic ischemic encephalopathy. Significant correlation was observed between plasma malondialdehyde and nitrate/nitrite levels with blood-brain barrier permeability.

CONCLUSIONS. Increased plasma levels of malondialdehyde and nitrates/nitrites are found to be associated with hypoxic ischemic encephalopathy, indicating the possible role of free radical injury in its causation. Increased blood-brain barrier permeability may be another contributory factor to the progression of the disease.
Comparing Nose-Throat Swabs and Nasopharyngeal Aspirates Collected From Children With Symptoms for Respiratory Virus Identification Using Real-Time Polymerase Chain Reaction
Authors: Lambert Stephen B. , Whiley David M. , O'Neill Nicholas T. , Andrews Emily C. , Canavan Fiona M. , Bletchly Cheryl , Siebert David J. , Sloots Theo P. , Nissen Michael D.
Published: August 2008

OBJECTIVES. The objective of this study was to calculate sensitivity values for the detection of major respiratory viruses of childhood by using combined nose-throat swabs and nasopharyngeal aspirates.

METHODS. Children who had symptoms and presented to a pediatric teaching hospital and had a diagnostic respiratory specimen collected were enrolled, and paired nose-throat swab and nasopharyngeal aspirate specimens were collected. Parents were asked to collect the nose-throat swab specimen in the first instance but could defer to a health care worker if unwilling. Nose-throat swab collectors were asked to rate perceived quality of collection. All nasopharyngeal aspirates were collected by a health care worker by using a standard protocol. Real-time polymerase chain reaction for 8 respiratory viruses was performed in our hospital's diagnostic laboratory.

RESULTS. Paired nose-throat swab/nasopharyngeal aspirate specimens were collected during 303 illnesses, with at least 1 respiratory virus identified in 186 (61%). For the major pathogens of childhood, influenza A virus and respiratory syncytial virus, collection by using the nose-throat swab had a sensitivity of 91.9% and 93.1%, respectively. A health care worker collected 219 (72%) of the nose-throat swab specimens; concordance with the nasopharyngeal aspirate was not related to health care worker collection or perceived quality of collection.

CONCLUSIONS. Nose-throat swab specimens, in combination with sensitive molecular testing, are a less invasive diagnostic respiratory specimen with adequate sensitivity for use in the clinic and hospital outpatient settings and large-scale community studies through parent collection. For children who present to a hospital in which an avian or pandemic strain of influenza virus is reasonably part of the differential diagnosis, nasopharyngeal aspirates or a similar collection technique (eg, nasal washes) should continue to be used.
Upper Social Strata Women in Nursing in Turkey
Authors: Özaydin Zuhal
Published: August 2008

The development of nursing education in Turkey was influenced by the twentieth-century political changes that encouraged the involvement of women in social life in Turkey. This study examines this development, beginning in the early twentieth century, including the role of relations between nurses in Turkey and the United States in advancing nursing education. The work is based on Ottoman archival sources, publications of the Ottoman-Turkish Red Crescent, and research on the history of nursing education in Turkey. The names of the institutions mentioned in documents and published works are in English, with the original Turkish names in parentheses. The dates in the Ottoman calendar (reckoned from the Hegira, Muslim era) and Roman calendar (adapted from the Gregorian calendar) that were used by Ottoman officials in their correspondence have been converted to the Western Christian calendar. English translations of Turkish references are in parentheses.
Frances U. Reiter and the Graduate School of Nursing at the New York Medical College, 1960–1973
Authors: Hiestand Wanda C.
Published: August 2008

In 1960, at the age of fifty-six, Frances U. Reiter (Figure 1) became dean of the newly established Graduate School of Nursing, New York Medical College, Flower and Fifth Avenue Hospitals. For Reiter, this bold venture in professional nursing education marked the attainment of a dream. Here at last was a graduate school of nursing requiring the baccalaureate degree for admission, under the direction of a nurse, and focused on education for advanced clinical practice. This paper tells the story of Reiter, the Graduate School of Nursing at New York Medical College, the development of an educational program for advanced nurse-clinician practice, and the belief that personal care of the patient is the heart of nursing.
Further Results on Asymmetry in Farm-Retail Price Transmission under Spatial Monopoly
Authors: Fousekis Panos
Published: August 2008

This work suggests that the range of parametric configurations which give rise to a strictly convex farm-retail price transmission function is wider than that identified by Azzam (1999). As a matter of fact, Azzam's implicit result (that is, a strictly convex aggregate demand function leads always to a strictly concave price transmission function) emerges only as a particular case of the result obtained here for large elasticity of aggregate demand and patient enough retailers.
Study of a circular Gaussian transition in an optical speckle field
Authors: Bergoënd Isabelle , Orlik Xavier , Lacot Eric
Published: August 2008

We propose a numerical approach to study optical speckle fields generated by various conditions of surface roughness and illumination, without any a priori assumption concerning the properties of the scattered field. By overcoming the limitations imposed by the use of the central limit theorem, we perform the study of the whole circular Gaussian transition undergone by the speckle field when varying the surface roughness and the size of the laser illumination. We focus our attention on the speckle contrast, the degree of circularity and the degree of asymmetry, including their radial dependence.
Entropic Behavior of Binary Carbonaceous Mesophases
Authors: Golmohammadi Mojdeh , Rey Alejandro D.
Published: August 2008

The Maier-Saupe model for binary mixtures of uniaxial discotic nematogens, formulated in a previous study [1], is used to compute and characterize orientational entropy [2] and orientational specific heat. These thermodynamic quantities are used to determine mixture type (ideal or non-ideal) which arise due to their different intrinsic properties, determined by the molecular weight asymmetry ΔMw and the molecular interaction parameter β. These molecular properties are also used to characterize the critical concentration where the mixture behaves like a single component system and exhibits the minimum nematic to isotropic (NI) transition temperature (pseudo-pure mixture). A transition within the nematic phase takes place at this specific concentration. According to the Maier-Saupe model, in a single mesogen, entropy at NI transition is a universal value; in this work we quantify the mixing effect on this universal property. The results and analysis provide a new tool to characterize molecular interaction and molecular weight differences in mesogenic mixtures using standard calorimetric measurements.
Calculation of Differential Entropy for a Mixed Gaussian Distribution
Authors: Michalowicz Joseph V. , Nichols Jonathan M. , Bucholtz Frank
Published: August 2008

In this work, an analytical expression is developed for the differential entropy of a mixed Gaussian distribution. One of the terms is given by a tabulated function of the ratio of the distribution parameters.
Incongruence between cpDNA and nrITS trees indicates extensive hybridization within Eupersicaria (Polygonaceae)
Authors: Kim Sang‐Tae , Donoghue Michael J.
Published: August 2008

Interspecific hybridization followed by polyploidization appears to have played a major role in plant diversification, but quantifying the contribution of this mechanism to diversification within taxonomically complex clades remains difficult. Incongruence among gene trees can provide critical insights, especially when combined with data on chromosome numbers, morphology, and geography. To further test our previous hypothesis on hybrid speciation in Persicaria (Polygonaceae), we performed molecular phylogenetic studies using three cpDNA regions and nuclear ITS sequences, with an emphasis on sampling within section Eupersicaria. Our analyses revealed major conflicts between the combined cpDNA tree and the nrITS tree; a variety of incongruence tests rejected stochastic error as the cause of incongruence in most cases. On the basis of our tree incongruence results and information on chromosome numbers, we hypothesize that the origin of 10 polyploid species involved interspecific hybridization. Our studies also support the recognition of several previously named species that have been treated as belonging within other species. Repeated allotetraploidy (as distinct from radiation at the tetraploid level) now appears to be the key mechanism governing the diversification of this taxonomically challenging group.
Hidden diversity of endophytic fungi in an invasive plant
Authors: Shipunov Alexey , Newcombe George , Raghavendra Anil K. H. , Anderson Cort L.
Published: August 2008

Fungal endophytes are important in plant ecology and common in plants. We attempted to test cointroduction and host‐jumping hypotheses on a community basis by comparing endophytes isolated from invasive spotted knapweed (Centaurea stoebe, Asteraceae) in its native and invaded ranges. Of 92 combined, sequence‐based haplotypes representing eight classes of Fungi, 78 occurred in only one of the two ranges. In the native range of C. stoebe, one haplotype of Alternaria alternata was clearly dominant, whereas in the invaded range, no haplotype was dominant. Many haplotypes were closely related to one another and novel. For example, six putative, new species of Botrytis were discovered as endophytes of C. stoebe, which has never been reported to have Botrytis spp.. Apparent differences between the two communities of endophytes were significant according to an analysis of similarity, but phylogenetic community structure did not differ significantly between the ranges. Both host‐jumping and cointroduction of fungal endophytes likely took place during the spotted knapweed invasion.
Host genotype overrides fungal endophyte infection in influencing tiller and spike production of Lolium perenne (Poaceae) in a common garden experiment
Authors: Cheplick Gregory P.
Published: August 2008

Leaves of many cool‐season grasses are infected by endophytic fungi that can impact their populations. A common garden experiment with Lolium perenne was established in a lawn in New Jersey, USA, to investigate the impact of endophyte infection and host genotype on tiller and spike production over three years. Infected (E+) and uninfected (E–) plants of each genotype were monitored every 2–3 mo. Infection intensity within plants varied significantly among genotypes and years, but there was no evidence of directional change over time. Tiller production varied significantly among genotypes and was affected by endophytes: E+ plants of several genotypes produced more tillers than E– plants during the third year. E+ plants had greater aboveground biomass, but host genotype explained a far greater proportion of variation in tiller production, number, and biomass than infection. Plant survival, percentage flowering, flowering date, number of spikes, and mean tiller mass were unaffected by endophytes. However, the last three variables showed significant variation among host genotypes. Although studies have demonstrated a positive growth effect of endophytes on several grass hosts, in this experiment host genotype accounted for far more of the variation in tiller and spike production and in biomass of Lolium perenne than endophyte infection.
In search of monophyletic taxa in the family Desmidiaceae (Zygnematophyceae, Viridiplantae): the genus Cosmarium
Authors: Gontcharov Andrey A. , Melkonian Michael
Published: August 2008

Nuclear‐encoded small subunit (SSU) rDNA, 1506 group I introns, and chloroplast rbcL genes were sequenced from 97 strains representing the largest desmid genus Cosmarium (45 spp.), its putative relatives Actinotaenium (5 spp.), Xanthidium (4 spp.), Euastrum (9 spp.), Staurodesmus (13 spp.), and other Desmidiaceae (Zygnematophyceae, Streptophyta) and used to assess phylogenetic relationships in the family. Analyses of single genes and of a concatenated data set (3260 nt) established 10 well‐supported clades in the family with Cosmarium species distributed in six clades and one nonsupported assemblage. Most of the clades contained representatives of at least two genera highlighting the polyphyletic nature of the genera Cosmarium, Euastrum, Staurodesmus, and Actinotaenium. To enhance resolution between clades, we extended the data set by sequencing the slowly evolving chloroplast‐encoded large subunit (LSU) rRNA gene from 40 taxa. Phylogenetic analyses of a concatenated data set (5509 nt) suggested a sister relationship between two clades that consisted mainly of Cosmarium species and included C. undulatum, the type species of the genus. We describe molecular signatures in the SSU rRNA for two clades and conclude that more studies involving new isolates, additional molecular markers, and reanalyses of morphological traits are necessary before the taxonomic revision of the genus Cosmarium can be attempted.
Toward a comprehensive understanding of phylogenetic relationships among lineages of Acanthaceae s.l. (Lamiales)
Authors: McDade Lucinda A. , Daniel Thomas F. , Kiel Carrie A.
Published: August 2008

Acanthaceae (Asteridae; Lamiales) include ∼4000 species and encompass a range of morphological diversity, habitats, and biogeographic patterns. Although they are important components of tropical and subtropical habitats worldwide, inadequate knowledge of the family's phylogenetic framework has impeded comparative research. In this study, we sampled all known lineages of Acanthaceae including Andrographideae. Also included were eight of 13 genera whose relationships remain enigmatic. We used sequence data from nrITS and four chloroplast noncoding regions, and parsimony and Bayesian methods of analysis. Results strongly support most aspects of relationships including inclusion of Avicennia in Acanthaceae. Excepting Neuracanthus, newly sampled taxa are placed with strong support; Kudoacanthus is in Justicieae, Tetramerium lineage, and the remaining enigmatic genera are in Whitfieldieae or Barlerieae, and Andrographideae are sister to Barlerieae. This last result is unanticipated, but placement of Andrographideae based on structural characters has been elusive. Neuracanthus is monophyletic but placement relative to (Whitfieldieae (Andrographideae + Barlerieae)) is weakly supported. Many clades have clear morphological synapomorphies, but nonmolecular evidence for some remains elusive. Results suggest an Old World origin with multiple dispersal events to the New World. This study informs future work by clarifying sampling strategy and identifying aspects of relationships that require further study.
Evolution and phylogenetic utility of the PHOT gene duplicates in the Verbena complex (Verbenaceae): dramatic intron size variation and footprint of ancestral recombination
Authors: Yuan Yao‐Wu , Olmstead Richard G.
Published: August 2008

A well‐resolved species level phylogeny is critically important in studying organismal evolution (e.g., hybridization, polyploidization, adaptive speciation). Lack of appropriate molecular markers that give sufficient resolution to gene trees is one of the major impediments to inferring species level phylogenies. In addition, sampling multiple independent loci is essential to overcome the lineage sorting problem. The availability of nuclear loci has often been a limiting factor in plant species‐level phylogenetic studies. Here the two PHOT loci were developed as new sources of nuclear gene trees. The PHOT1 and PHOT2 gene trees of the Verbena complex (Verbenaceae) are well resolved and have good clade support. These gene trees are consistent with each other and previously generated chloroplast and nuclear waxy gene trees in most of the phylogenetic backbone as well as some terminal relationships, but are incongruent in some other relationships. Locus‐specific primers were optimized for amplifying and sequencing these two loci in all Lamiales. Comparing intron size in the context of the gene trees shows dramatic variation within the Verbena complex, particularly at the PHOT1 locus. These variations are largely caused by invasions of short transposable elements and frequent long deletions and insertions of unknown causes. In addition, inspection of DNA sequences and phylogenetic analyses unmask a clear footprint of ancestral recombination in one species.
Hybridization and crossability inCaiophora(Loasaceae subfam. Loasoideae): Are interfertile species and inbred populations results of a recent radiation?
Authors: Ackermann Markus , Achatz Michaela , Weigend Maximilian
Published: August 2008

Interspecific hybridization is considered a possible mechanism of plant diversification. The Andes are a hotspot of biodiversity, but hybridization in Andean taxa has so far not been investigated intensively. The current study investigates crossability inCaiophora(Loasaceae subfam. Loasoideae) by experimental interspecific hybridization of seven different species. Hand pollination was undertaken, developing fruits counted, thousand (seed) grain weights, and seed viability were examined. Cross pollination led to some fruit set in 36 of the 37 different combinations. Overall fruit set was virtually identical irrespective of the combination of parental plants. Mean germination rates were much higher in hybrid seeds, indicating a marked heterosis effect and the possible presence of an inbreeding depression in the source populations: In experimental hybridization the divergent taxa ofCaiophorabehave like isolated, inbred populations of a single species. Allopatry and different habitat preferences seem to be the key factors keeping the (interfertile) taxa ofCaiophoraapart in the apparent absence of both postmating isolating mechanisms and obvious isolating mechanisms in phenology and floral biology. Interspecific hybrids reported from the wild appear to be the result of secondary contact due to human impact.
A Rat Primary Hepatocyte Culture Model for Aging Studies
Authors: Shenvi Swapna V. , Dixon Brian M. , Petersen Shay Kate , Hagen Tory M.
Published: August 2008

The purpose of this protocol is to establish a primary hepatocyte culture system as a suitable model to examine age‐related changes in Phase II detoxication gene expression. Hepatocytes are isolated using a two‐step collagenase perfusion technique from young (3 to 6 months) and old (24 to 28 months) rats and placed in primary culture using collagen (Type I)‐coated plates as the extracellular matrix. A supplemented William's E Medium is used as the medium. This culture system maintains hepatocyte viability from both young and old rats for ∼60 hr, as measured by lactate dehydrogenase activity, while also maintaining their respective phenotypes relative to Phase II detoxification. We thus conclude that a collagen‐based cell culture system is suitable to study age‐associated deficits in Nrf2/ARE‐mediated Phase II gene regulation provided that experiments can be conducted within 60 hr after cell isolation. Curr. Protoc. Toxicol. 37:14.7.1‐14.7.10. © 2008 by John Wiley & Sons, Inc.
Determination of Membrane Protein Molecular Weight Using Sedimentation Equilibrium Analytical Ultracentrifugation
Authors: Fleming Karen G.
Published: August 2008

Both the stoichiometry and stability of native membrane protein complexes pose challenges to understanding the biology of these proteins. Sedimentation equilibrium analytical ultracentrifugation is well recognized as a thermodynamically rigorous technique for determining these quantities for macromolecules. This unit describes the experimental strategies that can be used to extract this information for membrane proteins reconstituted in vitro in detergent micelle or detergent/lipid mixed micelle solutions. Curr. Protoc. Protein Sci. 53:7.12.1‐7.12.13. © 2008 by John Wiley & Sons, Inc.
Production of Polyclonal Antisera
Authors: Cooper Helen M. , Patterson Yvonne
Published: August 2008

Much of modern biology and biochemistry relies on the availability of highly specific antibodies for use in such ubiquitous techniques as immunohistochemistry, ELISAs, immunoprecipitation, and immunoblotting. Thus, the generation of large quantities of specific antibodies directed to proteins or peptides of interest is essential to the success of both basic and applied research programs. In addition, with the advent of antibody‐based proteomic strategies for profiling protein expression and post‐translational modification, a requirement for timely production of specific antibodies has emerged. Polyclonal antibodies derived from animals immunized with purified proteins or peptides are particularly valuable for use in the laboratory. This unit provides protocols for the production of polyclonal antisera specific for protein antigens in rabbits, rats, mice, and hamsters. Curr. Protoc. Immunol. 82:2.4.1‐2.4.10. © 2008 by John Wiley & Sons, Inc.
Measurement of Proliferative Responses of Cultured Lymphocytes
Authors: Muul Linda Mesler , Silvin Christopher , James Stephen P. , Candotti Fabio
Published: August 2008

Measurement of proliferative responses of human lymphocytes is a fundamental technique for the assessment of their biological responses to various stimuli. Most simply, this involves measurement of the number of cells present in a culture before and after the addition of a stimulating agent. This unit contains several different prototype protocols to measure the proliferative response of lymphocytes following exposure to mitogens, antigens, allogeneic or autologous cells, or soluble factors. Each of these protocols can be used in conjunction with an accompanying support protocol which contains methods for pulsing cultures with [3H]thymidine and determining incorporation of [3H]thymidine into DNA or assessing cell proliferation by nonradioactive methods, e.g., reduction of tetrazolium salts (MTT). The protocols described here provide an estimate of DNA synthesis and cell proliferation in an entire cell population, but do not provide information on the proliferation of individual cells. A protocol for CFSE labeling allows specific subpopulations of cells to be separated viably for further analysis. Curr. Protoc. Immunol. 82:7.10.1‐7.10.24. © 2008 by John Wiley & Sons, Inc.
Isolation and Visualization of Mouse Placental Hematopoietic Stem Cells
Authors: Gekas Christos , Rhodes Katrin E. , K.A. Mikkola Hanna
Published: August 2008

This unit describes the isolation of hematopoietic stem cells (HSCs) from the mouse placenta. The placenta was recently identified as an important hematopoietic site that generates HSCs de novo and provides a transitory niche for a large pool of HSCs during midgestation. This protocol includes a dissection technique for murine placenta, the mechanical and enzymatic steps of placental tissue dissociation, and phenotypical identification and isolation of HSCs. It also contains a method for immunohistochemical analysis of placenta tissue sections to visualize developing HSCs in the placenta. Curr. Protoc. Stem Cell Biol. 6:2A.1.1‐2A.1.14. © 2008 by John Wiley & Sons, Inc.
Generation of Cultured Oligodendrocyte Progenitor Cells from Rat Neonatal Brains
Authors: Larsen Eric C. , Kondo Yoichi , Fahrenholtz Cale D. , Duncan Ian D.
Published: August 2008

The oligodendrocyte progenitor cell (OPC) is one of the most studied progenitor cells of the body. It has been extensively researched in tissue culture and more recently in vivo using a wide range of markers that recognize transcription factors and cell surface markers and identify its earliest development from neural stem cells onward. Isolation of OPCs in large numbers and in purified preparations has been sought after as a source of cells for the repair of human myelin disorders. It has been proposed that such cells could be used as an exogenous source of cells for the treatment of lesions in multiple sclerosis and the less common genetic myelin disorders such as Pelizaeus‐Merzbacher disease. Prior to clinical trials, such approaches can be tested in animal models. Here, we describe the isolation of rat OPCs in culture conditions that provide large numbers of purified populations of cells. Curr. Protoc. Stem Cell Biol. 6:2D.1.1‐2D.1.13. © 2008 by John Wiley & Sons, Inc.
Preparing for Biofilm Studies in the Field
Authors: McLean Robert J.C. , Simpson Thomas R.
Published: August 2008

In their natural environments, microorganisms are under constant environmental selection to form biofilms. Using aquatic biofilms as an example, this unit illustrates general concepts in field biology and practical suggestions for designing, conducting, and analyzing biofilm experiments at varying distances from the laboratory. The unit also addresses an example of a special situation (space flight) where experimentation must be done by proxy through another individual or machine.
Laboratory Maintenance of Bartonella quintana
Authors: Battisti James M. , Minnick Michael F.
Published: August 2008

Trench fever is the common name for the acute febrile syndrome associated with a Bartonella quintana bacterial infection. The focus of this unit is to describe reliable methods for cultivation and cryopreservation of B. quintana and can be applied to cultivation and preservation of all Bartonella. Detailed recipes for preparation of three types of semisolid media are also included.
Growth and Laboratory Maintenance of Campylobacter jejuni
Authors: Davis Lindsay , DiRita Victor
Published: August 2008

Campylobacter jejuni is a fastidious organism, growing in microaerophilic conditions with a temperature range between 37° and 42°C. Multiple types of media can be used to cultivate it; however, Mueller Hinton broth and agar support the best C. jejuni growth. Optimum atmosphere for C. jejuni is 85% N2, 10% CO2, and 5% O2. Curr. Protoc. Microbiol. 10:8A.1.1‐8A.1.7. © 2008 by John Wiley & Sons, Inc.
Animal Models of Listeria Infection
Authors: Cabanes Didier , Lecuit Marc , Cossart Pascale
Published: August 2008

Listeria monocytogenes is an intracellular foodborne pathogen that causes listeriosis, an infection characterized by gastroenteritis, meningitis, encephalitis, and maternofetal infections in humans. L. monocytogenes enters the host via contaminated foods, invades the small intestine, translocates to mesenteric lymph nodes, and spreads to the liver, spleen, brain and, in pregnant women, the fetoplacental unit. Many pathogenicity tests for studying L. monocytogenes have been developed, including tests using laboratory animals. A number of small animal species can be experimentally infected with Listeria. Mice and guinea pigs can be infected either intragastrically or intravenously, and virulence evaluated either by enumerating bacteria within infected target organs or by evaluating the 50% lethal dose (LD50). Although mice and guinea pigs can be infected with Listeria by a variety of routes, the intragastric route is the most relevant to the human foodborne listeriosis.
Aphid Transmission of Plant Viruses
Authors: Gray Stewart M.
Published: August 2008

A majority of plant viruses are transmitted between hosts by insect vectors, and it is often important to use insect transmission in the laboratory to maintain virus isolates or to study virus‐vector‐plant interactions. Although many of these viruses can also be mechanically transmitted in the laboratory using infected sap, maintenance by mechanical transmission can often lead to changes in the virus, either minor changes in gene sequences or, in some cases, major deletions of genome sequences. These can affect both virus‐vector and virus‐host interactions. This unit describes some simple and practical methods for conducting virus transmission experiments using sap‐sucking insects. Curr. Protoc. Microbiol. 10:16B.1.1‐16B.1.10. © 2008 by John Wiley & Sons, Inc.