bluuebunny/crossref_metadata_2025 huggingface dataset:
Multifunctional factor ENY2 is associated with the THO complex and promotes its recruitment onto nascent mRNA
Authors: Kopytova Daria V. , Orlova Anastasija V. , Krasnov Alexey N. , Gurskiy Dmitriy Ya. , Nikolenko Julia V. , Nabirochkina Elena N. , Shidlovskii Yulii V. , Georgieva Sofia G.
Published: January 2010

Metazoan E(y)2/ENY2 is a multifunctional protein important for transcription activation and mRNA export, being a component of SAGA/TFTC and the mRNA export complex AMEX. Here, we show that ENY2 in Drosophila is also stably associated with THO, the complex involved in mRNP biogenesis. The ENY2–THO complex is required for normal Drosophila development, functioning independently on SAGA and AMEX. ENY2 and THO arrive on the transcribed region of the hsp70 gene after its activation, and ENY2 plays an important role in THO recruitment. ENY2 and THO show no direct association with elongating RNA polymerase II. Recruitment of ENY2 and THO occurs by their loading onto nascent mRNA, apparently immediately after its synthesis, while the AMEX component Xmas-2 is loaded onto mRNA at a later stage. Knockdown of either ENY2 or THO, but not SAGA or AMEX, affects the processing of the transcript's 3′ end. Thus, ENY2, as a shared subunit of several protein complexes governing the sequential steps of gene expression, plays an important role in the coordination of these steps.
C/EBPβΔuORF mice—a genetic model for uORF-mediated translational control in mammals
Authors: Wethmar Klaus , Bégay Valérie , Smink Jeske J. , Zaragoza Katrin , Wiesenthal Volker , Dörken Bernd , Calkhoven Cornelis F. , Leutz Achim
Published: January 2010

Upstream ORFs (uORFs) are translational control elements found predominantly in transcripts of key regulatory genes. No mammalian genetic model exists to experimentally validate the physiological relevance of uORF-regulated translation initiation. We report that mice deficient for the CCAAT/enhancer-binding protein β (C/EBPβ) uORF initiation codon fail to initiate translation of the autoantagonistic LIP (liver inhibitory protein) C/EBPβ isoform. C/EBPβΔuORF mice show hyperactivation of acute-phase response genes, persistent repression of E2F-regulated genes, delayed and blunted S-phase entry of hepatocytes after partial hepatectomy, and impaired osteoclast differentiation. These data and the widespread prevalence of uORFs in mammalian transcriptomes suggest a comprehensive role of uORF-regulated translation in (patho)physiology.
The Rabbit(Oryctolagus cuniculus):A Model for Mammalian Reproduction and Early Embryology
Authors: Püschel Bernd , Daniel Nathalie , Bitzer Eva , Blum Martin , Renard Jean-Paul , Viebahn Christoph
Published: January 2010

INTRODUCTION

The rabbit is a valuable animal model for a variety of biomedical research areas including in vitro fertilization, early embryology and organogenesis, neurophysiology, ophthalmology, and cardiovascular research. The rabbit is also used as a model for toxicology studies and analyses of drug effects on embryo and fetal development, as well as for research involving the immune system (e.g., host/pathogen interactions, asthma, arthritis, systemic lupus erythematosus). The rabbit’s most prominent use as a laboratory animal is antibody production; most biomedical research institutions thus provide the infrastructure and veterinary expertise for keeping rabbits. This article discusses current advances in applying modern life science research methods to basic and applied embryological research using the rabbit as a model for early intrauterine development in mammals, including state-of-the-art genomic tools and the sequencing data available (and planned) for the rabbit.
Phenotypic Analysis of Arabidopsis Mutants: Electrolyte Leakage after Freezing Stress
Authors: Lee Byeong-ha , Zhu Jian-Kang
Published: January 2010

INTRODUCTION

Abiotic stress such as high salt or low temperature adversely affects plant growth and development. Salt stress inhibits seed germination, retards plant growth, and accelerates senescence. Freezing or drought stress can cause cell damage and plant death. The following parameters can be used to evaluate plant tolerance to salt, drought, or freezing stress: root elongation, fresh weight gain, seed germination, electrolyte leakage (described here), or water-loss measurement. Several stress mutants have been characterized using these tests, including hos1 and hos2, which show higher expression of some stress-regulated genes when exposed to low-temperature stress; hos5, which shows higher expression of some stress-regulated genes under abscisic acid (ABA) and salt treatments; sfr mutants, which are deficient in freezing tolerance; and eskimo1, which is constitutively freezing tolerant. This protocol describes an electrolyte leakage assay that can be used to measure the degree of cell damage after freezing stress in Arabidopsis. The levels of stress suggested in this protocol may need to be adjusted, depending on the ecotype and growth conditions used.
Dissection and Fixation of Rabbit Embryos
Authors: Püschel Bernd , Viebahn Christoph
Published: January 2010

INTRODUCTION

This protocol describes general methods for dissection and fixation of rabbit embryos. Depending on the investigation, embryos can be dissected either alive or after fixation. Some embryonic stages are surrounded by an envelope, the zona pellucida, which might have to be removed for some applications (e.g., immunohistochemistry). Fixation methods vary according to different experimental needs, such as structure preservation (morphology), antigen preservation (immunoreactivity), or mRNA accessibility (in situ hybridization).
Whole-Mount In Situ Hybridization of Early Rabbit Embryos
Authors: Püschel Bernd , Blum Martin , Viebahn Christoph
Published: January 2010

INTRODUCTION

This protocol describes whole-mount in situ hybridization of early rabbit embryos. The method allows specific mRNA expression to be localized with respect to position and intensity in predefined embryonic areas. Optimal resolution of mRNA distribution can be achieved by subsequently embedding in situ hybridized embryos for histological sectioning.
Artificial Fertilization in Rabbits by Intracytoplasmic Sperm Injection (ICSI): Figure 1.
Authors: Daniel Nathalie , Renard Jean-Paul
Published: January 2010

INTRODUCTION

Intracytoplasmic sperm injection (ICSI) consists of the injection, under microscopic observation, of a single spermatozoon directly into the cytoplasm of an oocyte using an injection capillary. This technique is used commonly in human medicine in cases of infertility. The advantage of ICSI is that the technique requires only one spermatozoon per oocyte for fertilization. Spermatozoa used for ICSI do not require high motility; ICSI works as long as the sperm nucleus is genetically intact. This protocol describes methods for performing ICSI in rabbits using either fresh or frozen/thawed sperm.
Cryopreservation of Rabbit Embryos
Authors: Daniel Nathalie , Renard Jean-Paul
Published: January 2010

INTRODUCTION

In the rabbit, the technique most frequently used for embryo transfer involves cryopreservation by slow freezing of morulae, followed by surgical implantation of thawed embryos. This method has the advantage of being well controlled and validated: It helps to maintain mitochondrial genomes, preserves the genetic integrity of biological material under sanitary conditions, and allows for such material to be passed along beyond the normal reproductive lifespan of a given animal, which can be advantageous in the case of rare or valuable strains. Furthermore, because litters of pups can be obtained 1 mo after embryo transfer without the need for additional backcrossings, genotyping is rapid and simple. However, the technique is not without its drawbacks: It requires expensive equipment (e.g., specialized programmable freezers), and producing large numbers of stored embryos requires the sacrifice of many donor females. Although alternative methods of embryo recovery (e.g., surgical or vaginal collection) are possible, such procedures produce fewer embryos.
Maintaining Clonal Paramecium tetraurelia Cell Lines of Controlled Age through Daily Reisolation
Authors: Beisson Janine , Bétermier Mireille , Bré Marie-Hélène , Cohen Jean , Duharcourt Sandra , Duret Laurent , Kung Ching , Malinsky Sophie , Meyer Eric , Preer John R. , Sperling Linda
Published: January 2010

INTRODUCTION

The sexual cycle of Paramecium tetraurelia can be managed by controlling food uptake, allowing the study of developmentally regulated differentiation in synchronous cultures. Clonal cell lines can be established easily by single cell isolation and can be grown in bacterized grass infusion medium for a large number of vegetative divisions before entering senescence. Sexual reproduction is triggered by starvation and therefore can be induced or prevented as needed by controlling the availability of food. Given a constant supply of food, P. tetraurelia cells will remain in the vegetative phase of their life cycle and, at 27°C, will divide by binary fission every 6 h. The daily reisolation procedure described here results in populations of P. tetraurelia of known age and produced under reproducible physiological conditions, which can help to standardize experiments. Paramecia produced under these conditions are suitable for immunocytochemical studies or use in genetic cross experiments.
Fluorescence In Situ Hybridization for the Identification of Environmental Microbes
Authors: Wendeberg Annelie
Published: January 2010

INTRODUCTION

This protocol presents methods for the phylogenetic identification of microorganisms in environmental samples (e.g., water and sediments) by means of fluorescence in situ hybridization (FISH) with rRNA-targeted oligonucleotide probes, followed by signal amplification with catalyzed reporter deposition (CARD). In this procedure, FISH probes are conjugated with the enzyme horseradish peroxidase (HRP). After hybridization, the subsequent deposition of fluorescently labeled tyramides results in substantially higher signal intensities on target cells than after FISH with probes labeled directly with fluorochromes. This protocol describes the custom labeling of tyramides with different fluorochromes, as well as sample preparation and cell permeabilization strategies for various microbial cell wall types. A protocol for sequential multicolor CARD-FISH for the simultaneous detection of different phylogenetic groups is also presented.
Cryosectioning and Immunocytochemistry of Adult Drosophila Eye Sections
Authors: Wolff Tanya
Published: January 2010

INTRODUCTION

This protocol is recommended for antibody staining of frozen adult Drosophila eye sections. It describes the tissue preparation and staining with primary and secondary antibodies.
Meta‐analysis: targeting the intestinal microbiota in prophylaxis for post‐operative Crohn’s disease
Authors: DOHERTY G. A. , BENNETT G. C. , CHEIFETZ A. S. , MOSS A. C.
Published: January 2010

Aliment Pharmacol Ther 31, 802–809

Background  Enteric bacteria play an important early role in the pathogenesis of Crohn’s disease.

Aim  To perform a meta‐analysis of trials testing antibiotics or probiotics for prevention of post‐operative recurrence of Crohn’s disease.

Methods  Review of all randomized controlled trials comparing antibiotics or probiotics with placebo in prevention of endoscopic or clinical recurrence of Crohn’s disease after surgical resection. Fixed‐effect meta‐analysis was performed with dichotomous data summarized using relative risk with 95% confidence intervals, where appropriate.

Results  Seven studies were identified as suitable for inclusion (two comparing antibiotics with placebo, five comparing probiotics with placebo). The use of nitroimidazole antibiotics (metronidazole, ornidazole) reduced the risk of clinical (RR 0.23; 95% CI 0.09–0.57, NNT = 4) and endoscopic (RR 0.44; 95% CI 0.26–0.74, NNT = 4) recurrence relative to placebo. However, these agents were associated with higher risk of adverse events (RR 2.39, 95% CI 1.5–3.7) and patient withdrawal. Probiotic administration was not associated with any significant difference in risk of recurrence compared with placebo.

Conclusions  Nitroimidazole antibiotics are effective in the prevention of post‐operative Crohn’s disease recurrence, but their side‐effects limit acceptability. Probiotics have failed to show efficacy for post‐operative prophylaxis, but may merit further study.
The pattern of linkage disequilibrium in German Holstein cattle
Authors: Qanbari S. , Pimentel E. C. G. , Tetens J. , Thaller G. , Lichtner P. , Sharifi A. R. , Simianer H.
Published: January 2010

This study presents a second generation of linkage disequilibrium (LD) map statistics for the whole genome of the Holstein–Friesian population, which has a four times higher resolution compared with that of the maps available so far. We used DNA samples of 810 German Holstein–Friesian cattle genotyped by the Illumina Bovine SNP50K BeadChip to analyse LD structure. A panel of 40 854 (75.6%) markers was included in the final analysis. The pairwise r2 statistic of SNPs up to 5 Mb apart across the genome was estimated. A mean value of r2 = 0.30 ± 0.32 was observed in pairwise distances of <25 kb and it dropped to 0.20 ± 0.24 at 50–75 kb, which is nearly the average inter‐marker space in this study. The proportion of SNPs in useful LD (r2 ≥ 0.25) was 26% for the distance of 50 and 75 kb between SNPs. We found a lower level of LD for SNP pairs at the distance ≤100 kb than previously thought. Analysis revealed 712 haplo‐blocks spanning 4.7% of the genome and containing 8.0% of all SNPs. Mean and median block length were estimated as 164 ± 117 kb and 144 kb respectively. Allele frequencies of the SNPs have a considerable and systematic impact on the estimate of r2. It is shown that minimizing the allele frequency difference between SNPs reduces the influence of frequency on r2 estimates. Analysis of past effective population size based on the direct estimates of recombination rates from SNP data showed a decline in effective population size to Ne = 103 up to ∼4 generations ago. Systematic effects of marker density and effective population size on observed LD and haplotype structure are discussed.
Intranidal mating, gyne polymorphism, polygyny, and supercoloniality as factors for sympatric and parapatric speciation in ants
Authors: SEIFERT BERNHARD
Published: January 2010

1. The syndrome of intranidal mating, morpho‐ethological gyne polymorphism, and polygyny is found in less than 5% of European ant species, but is considered as an important precondition for sympatric and parapatric speciation.

2. This idea is discussed in three cases that represent example of evolution below the species level: (1) the polygynous—polydomous form of Lasius turcicus as a model of how speciation of the supercolonial Lasius neglectus could have taken place; (2) the microgyne form of Myrmica rubra as a model for sympatric speciation of socially parasitic species; and (3) hybridisation and rapid genotype selection in supercolonial wood ants of the Formica rufa group as a model for hybrid speciation.

3. The first step of the evolutionary scenario is transition to polygyny followed by development of morpho‐ethological gyne polymorphism: smaller gynes with weaker wing muscles and body reserves shift to intranidal mating and reduce dispersal flight, while larger gynes with bigger wing muscles and body reserves retain nuptial and dispersal flight. This scenario leads to mating place separation and facilitates further genotype divergence.

4. There is much less gene flow between conspecific supercolonies of Formica rufa group ants with dominance of intranidal or intracolonial mating than between conspecific monogynous colonies performing a nuptial flight. If two supercolonial species occasionally hybridise and if landscape structure provides some isolation of the resulting hybrid supercolony, very strong selection for genotypes takes place within this cohesive system of thousands of reproductive females eventually leading to novel genotypes and new species.
Shift happens! Shifting balance and the evolution of diversity in warning colour and mimicry
Authors: MALLET JAMES
Published: January 2010

1. At first sight, it seems most unlikely thatHeliconiuswarning colour races have evolved by means of stochastic peak shift or shifting balance.

2. Phase I, random local processes (including genetic drift and idiosyncratic selection), and phase III, interdemic selection are the most controversial phases of the shifting balance. Phase II consists of ordinary natural selection to a new adaptive peak within populations, and is uncontroversial.

3.Heliconiushave bold patterns of iridescent blue, black, yellow, white, and red. These are clearly warning patterns, and near‐perfect Müllerian mimicry has evolved among species, suggesting tight control by natural selection. Field experiments have also demonstrated strong selection (oftens >0.1 on single colour pattern loci), and the population structure ofHeliconiusis typically not conducive to phase I. Yet the colour patterns are clearly somewhat independent and incompatible signals of unpalatability.

4. There is empirical evidence inHeliconiusfor both controversial phases. For phase I, occasional and local polymorphisms of colour pattern in a number of species go against the generally expected (and generally observed) monomorphism for Müllerian mimics.

5. For phase III, one of the few colour pattern clines mapped in detail has been observed to move rapidly over a period of 20 years. There are also a number of curious ‘leapfrog’ geographical disjunctions in colour pattern races. Disjunctions are expected if successful races have spread from the centre of the range (e.g. the Amazonian rayed races) via phase III, in competition with earlier races that are now distributed in scattered places along the periphery of the range.

6. Evidence from the genomes ofHeliconiusmay in the near future aid in understanding colour pattern ‘supergenes’ and to help test for origin and spread via shifting balance.
The roles and interactions of reproductive isolation mechanisms in fall armyworm (Lepidoptera: Noctuidae) host strains
Authors: GROOT ASTRID T. , MARR MELANIE , HECKEL DAVID G. , SCHÖFL GERHARD
Published: January 2010

1. The moth Spodoptera frugiperda presents an interesting opportunity to study the evolution of reproductive isolation, because it consists of two host races that may be in the process of speciation.

2. The two races exhibit habitat isolation through host‐plant preference, and two types of behavioural isolation, i.e. differences in sex pheromone composition and timing of mating activity at night.

3. In this paper, we review the selection pressures acting upon these three barriers as well as their genetic bases, to address the question of how divergence of the two strains may have evolved.

4. We also address possible interactions between the three barriers, whether and how they may have evolved in concert, and we view the evolution of these three prezygotic isolation barriers in the light of postzygotic isolation.
Setting the scene… meeting up with Darwin and Wallace
Authors: LOXDALE HUGH D.
Published: January 2010

1. In 1858 the theory of evolution by means of natural selection was born. By the time of the publication of the two papers by Charles Robert Darwin (1809–1882) and Alfred Russel Wallace (1823–1913), Darwin was already a well established and respected scientist who had been thinking about evolution for a long time, at least since his world voyage in HMS Beagle from 1831 to 1836. In contrast, as far as science and the general public was then concerned, Wallace was comparatively unknown and largely an outsider. Yet he too independently developed a theory of evolution involving the mechanism of natural selection, following his travels to the Far East in the years 1854–1862 to collect and observe animal and, to a lesser extent, plant life. It could thus be argued that this was a theory whose time had come, and perhaps others would have developed such a theory shortly afterwards, certainly by the end of the 19th century. However, Darwin and Wallace got there first, especially Darwin who in the following year, 1859, published his magnum opus–On the Origin of Species. Their convincing and courageous theoretical explanation of how nature adaptively radiates and evolves under the influence of natural selection is now generally accepted by most biologists and many others worldwide. As such, it provides a logical construct of how organisms develop, compete, survive within established ecological niches, and adapt to new ones… or not as the case may be, whereupon individuals, populations, and indeed whole taxonomic groups, become extinct over time.
Rapid genetic changes in natural insect populations
Authors: LOXDALE HUGH D.
Published: January 2010

1. The insects represent around 75% of the world's fauna and as such provide especially good examples of the evolutionary process in action, aided by their often rapid generation time and high rate of reproduction.

2. Here, I review some of the main mechanisms of mutational, ecological, and evolutionary change in insects. All those described, whether allo‐ para‐ or sympatric, involve changes in the genome or in behaviour that may ultimately isolate newly changed individuals from the parental population/s. They include: loss of sexuality by various means, including (potentially) mutation of the gene/s controlling sexuality; karyotypic changes, both in terms of the number of chromosomes, translocation, polyploidy, and hybridisation; host shifts as pre‐ and post‐zygotic isolating mechanisms, and in asexuals such as aphids, ‘divergence hitchhiking’ around key quantitative trait loci (QTL), and in moths, selection acting at a few linkage groups; enzyme‐based adaptive changes; sex and contact pheromone‐based isolating mechanisms; phenotypic plastic changes; and epigenetic changes. These last such changes are brought about by stress inducers, and may be transgenerational in their effects.

3. Of the above mechanisms, probably chromosomal changes and host shifts represent the commonest mechanisms.
Reinforcement in hybrids among once isolated semispecies ofDanaus chrysippus(L.) and evidence for sex chromosome evolution
Authors: SMITH DAVID A. S. , GORDON IAN J. , ALLEN JOHN A.
Published: January 2010

1. The aim of this paper is to investigate mechanisms of reinforcement between two semi‐isolated semispecies of the African savannah butterflyDanaus chrysippus. The biogeography of colour genes suggests that four semispecies evolved in once isolated refugia. They expanded their ranges in response to Holocene climatic changes to form a hybrid zone in central‐east Africa.

2.Danaus chrysippusis a superspecies within which cycles of alternating cladogenesis and reticulation among semispecies have probably operated over some 4 Myr. Semispecies are inter‐fertile but show Haldane rule effects in crosses; gene flow is massive but subject to isolation by distance.

3. One semispecies shows linkage disequilibrium, vis‐ à‐vis others, for haplotype, karyotype (W‐linkage of colour genes which function as reproductive isolating barriers) and all‐female broods caused by a male‐killer endosymbiont. Introgression of colour genes betweenD. c. dorippusandD. c. chrysippusis constrained by sex linkage and male killing.

4. Reinforcement in hybrid zones comprises allochronic migration, assortative mating, (assumed) sex chromosome incompatibility and sex‐ratio distortion. Gene introgression fromD. c. dorippusto other semispecies is maintained by a high frequency of backcrossing between hybrid males and females of the latter.
Sequential speciation and the diversity of parasitic insects
Authors: FEDER JEFFREY L. , FORBES ANDREW A.
Published: January 2010

A major issue in biology is what generates biodiversity. An important part of the answer may be biodiversity itself. As new species form they may often create new niches, providing an impetus for other organisms to adapt and speciate. This hypothesis of ‘cascading’ or ‘sequential’ speciation has been argued to be particularly relevant for host‐plant‐specific phytophagous insects and their associated parasites. Here, we discuss the empirical and theoretical bases for sequential speciation and its potential significance for understanding the genesis of new life forms. We examine the application of the concept that biodiversity begets biodiversity by palaeontologists, ecologists, and evolutionary biologists to explain current and past distributions of organisms. We look at factors that may promote and restrict sequential speciation and review evidence from phytophagous insects for the process in nature. In particular, we discuss a case study involving the apple maggot fly, Rhagoletis pomonella (Diptera: Tephritidae) and its wasp parasitoid Diachasma alloeum (Hymenoptera: Braconidae). We conclude by outlining a research agenda for testing for sequential speciation in the field.
Micro‐evolutionary change in relation to insecticide resistance in the peach–potato aphid,Myzus persicae
Authors: FENTON BRIAN , MARGARITOPOULOS JOHN T. , MALLOCH GAYNOR L. , FOSTER STEPHEN P.
Published: January 2010

1. Phenotypic diversity is the fuel that powers evolution.

2. Asexual organisms rely on mutation whereas sexual organisms combine mutation with recombination.

3. Few organisms provide examples of species that are both sexual and asexual, but aphids do.

4. To examine evolution on perceptible timescales requires strong evolutionary forces and, as Darwin noted, agricultural practices provide strong selection. In the case of aphids, insecticides provide a considerable force in the elimination of genotypes.

5. Insecticide resistance inMyzus persicae(Sulzer) has arisen independently through point mutation and gene amplification on a number of occasions and at different times. Resistance to organophosphates, pyrethroids, and pirimicarb (a dimethyl carbamate) is now widespread.

6. In this paper, we examine these three elements: sexual recombination, clonal expansion, and insecticide selection in the peach–potato aphidM. persicaein relation to the evolution of insecticide resistance and survival of the fittest clone.
Longer‐acting factor VIII to overcome limitations in haemophilia management: the PEGylated liposomes formulation issue
Authors: DI MINNO G. , CERBONE A. M. , COPPOLA A. , CIMINO E. , DI CAPUA M. , PAMPARANA F. , TUFANO A. , DI MINNO M. N. D.
Published: January 2010

Summary.  Injected factor VIII (FVIII), the current treatment for haemophilia A, leads to major improvements in the quality of life and life expectancy of individuals with this disorder. However, because injected FVIII has a short half‐life in vivo, this strategy has major limitations for highly demanding regimens (e.g. prophylaxis, immune tolerance induction, surgery). Newer formulations of longer‐acting FVIII are presently under investigation. The use of low molecular weight polyethylene glycol (PEG)‐containing liposomes as carriers for recombinant FVIII (rFVIII) results in the prolongation of haemostatic efficacy. Data from preclinical experiments in mice, early clinical evaluations, and pharmacokinetics and pharmacodynamics results indicate that an rFVIII pegylated liposomal formulation may provide potential clinical benefit to patients with severe haemophilia A by prolonging the protection from bleeding. In light of this potential clinical benefit, a multicentre, randomized, active‐controlled, non‐inferiority phase II trial with two parallel treatment arms and equal randomization after stratification for the presence or absence of target joints in patients and for ages ≥18 years vs. <18 years is currently being conducted. The study will test the hypothesis that rFVIII‐Lip once‐weekly prophylaxis is not inferior to rFVIII‐water for injection thrice‐weekly prophylaxis. A total of 250 patients will be enrolled with severe haemophilia A (<1% FVIII) on on‐demand or secondary prophylaxis treatment and with documented bleeds or injections during the 6 months before study entry. Sixty‐four centres in 14 different countries are involved in the study; recruitment is underway. In Italy, six centres have already included 15 patients (no screening failure). Eight of these patients have completed the run‐in phase and have begun the home treatment. No unexpected serious adverse events have been reported thus far. Data emerging from this phase II study will help collect relevant data to overcome current limitations in haemophilia management by employing treatment with longer‐acting rFVIII.
The use of central venous catheters in haemophilia patients
Authors: IZZI G. , FRANCHINI M. , BONETTI L. , TAGLIAFERRI A.
Published: January 2010

Summary.  Central venous access devices (CVADs) play an important role in the management of haemophilia patients requiring repeated and/or urgent administration of coagulation factor concentrates. In this article, we summarize current knowledge regarding the use of central venous catheters in these patients, indicating advantages and disadvantages of both fully implantable and external tunnelled CVADs. Finally, we describe our personal experience on the use of the external tunnelled catheter Broviac.
Understanding inhibitor development in haemophilia A: towards clinical prediction and prevention strategies
Authors: COPPOLA A. , SANTORO C. , TAGLIAFERRI A. , FRANCHINI M. , DI MINNO G.
Published: January 2010

Summary.  Inhibitor development, because of its impact on patients’ morbidity and quality of life, is presently the most serious complication of haemophilia A treatment. The identification of several genetic and non‐genetic risk factors may be used for the stratification of inhibitor risk and the definition of prevention strategies, particularly for patients with a high‐risk genetic profile. The most extensively studied genetic factor is the type of F8 mutation, i.e. large deletions, nonsense mutations and inversions, which are associated with a higher risk of inhibitor development. This is the basis for the increased risk in patients with inhibitor family history; however, concordance family studies showed that factors other than F8 mutations are involved. An emerging role is investigated for polymorphisms of immune‐regulatory genes that may increase (IL‐10 and TNF‐α) or reduce (CTLA‐4) inhibitor risk and whose heterogeneous ethnic distribution may correlate to the higher inhibitor risk in non‐caucasian patients. A role for FVIII haplotypes, particularly in black haemophiliacs, has been recently proposed. Recent studies report an increased inhibitor risk for initial intensive treatments (surgery or severe bleeds requiring high‐dose and/or prolonged treatment, presence of danger signals), whereas regular prophylaxis (absence of danger signals) exerts a protective effect. A clinical score including the type of F8 mutation, family history of inhibitors and intensive treatment has been recently validated for predicting inhibitor risk. Because of the lack of useful data regarding the role of different types of FVIII concentrates, the stratification of risk in patients starting replacement treatment together with the careful evaluation of indications, doses and duration of treatment at first exposures and further efforts for overcoming barriers to early implementation of prophylaxis are encouraged, particularly for patients with a predictable high inhibitor risk.
In vivo transfer of plasmid pRAS1 between Aeromonas salmonicida and Aeromonas hydrophila in artificially infected Cyprinus carpio L.
Authors: Bello‐López J M , Fernández‐Rendón E , Curiel‐Quesada E
Published: January 2010

This study investigated the possible in vivo transfer of plasmid pRAS1 between Aeromonas salmonicida and A. hydrophila inhabiting two different organs of Cyprinus carpio L. To distinguish transconjugants from naturally occurring antibiotic resistant bacteria, twelve luminescent transposon‐tagged A. hydrophila strains using miniTn5luxCDABEKm2 transposon were generated. In conjugal transfer experiments, fish were conditioned with the donor bacteria and subsequently immersed in water containing the recipient strain. Bacteria were recovered from gills and intestines and isolated by growth on selective plates. Transconjugants were identified by their resistance to the pRAS1 encoded antimicrobials and by light emission. In vivo transfer frequencies ranged between 10−3 and 10−6 and were somewhat lower in intestines, compared to gills. Transfer frequencies were also smaller relative to those obtained in vitro. The minimal amount of donor and recipient bacteria needed to yield detectable transconjugants in vivo was 1 × 104 CFU mL−1. Implications of this plasmid transfer in natural settings and its possible consequences to human health are discussed.
A study of gross, histological and blood biochemical changes in rainbow trout, Oncorhynchus mykiss (Walbaum), with rainbow trout gastroenteritis (RTGE)
Authors: Del‐Pozo J , Turnbull J F , Crumlish M , Ferguson H W
Published: January 2010

The mechanisms behind the pathogenesis of rainbow trout gastroenteritis (RTGE) are still unknown. This study examined the macroscopic and microscopic changes in trout with RTGE (RTGE+), as well as the blood chemistry. A total of 464 rainbow trout were sampled from 11 sites in the UK, comprising 152 RTGE+ fish and 330 random, apparently healthy fish. A case definition for RTGE was assessed by the analysis of its agreement with three laboratory tests: histopathology, packed cell volume and kidney bacteriology. Cluster analysis indicated the presence of three distinct presentations within the population of RTGE+ fish. Cluster A included gross signs associated with moribund RTGE+ fish, and clusters B and C identified gross signs consistent with concurrent diseases, notably furunculosis, enteric redmouth and proliferative kidney disease. The information gained was used to select RTGE+ fish without concurrent disease for the analysis of RTGE pathogenesis with blood biochemistry. This analysis revealed a severe osmotic imbalance and a reduced albumin/globulin ratio as indicatives of selective loss of albumin. These findings are compatible with a protein losing enteropathy.
Detection of infectious pancreatic necrosis virus (IPNV) from asymptomatic redbanded seabream, Pagrus auriga Valenciennes, and common seabream, Pagrus pagrus (L.), using a non‐destructive procedure
Authors: Lopez‐Jimena B , Garcia‐Rosado E , Infante C , Cano I , Manchado M , Castro D , Borrego J J , Alonso M C
Published: January 2010

A non‐destructive procedure based on nested RT‐PCR and dot‐blot hybridization has been developed for the detection of asymptomatic IPNV‐carrier fish. The pair of primers designed for RT‐PCR amplified a 599‐bp fragment of the pVP2 region within the polyprotein gene, resulting in the detection of IPNV genotype III.1. The use of a nested RT‐PCR allowed the amplification of IPNV genotypes III.1 and I.2. In addition, a 191‐bp probe was designed for hybridization studies used in combination with the nested RT‐PCR. The application of the nested RT‐PCR to analyse blood samples from asymptomatic redbanded seabream, Pagrus auriga, and common seabream, P. pagrus, specimens showed a 53.1% and 77.8% prevalence of IPNV‐carriers, respectively. The combination of nested RT‐PCR and dot‐blot hybridization increased the detection rates up to 100% for redbanded seabream and 94.4% for common seabream. Therefore, the protocol described in this study is highly sensitive and specific for the detection of IPNV in asymptomatic carrier fish, and, in addition, the results demonstrate the carrier state in two newly cultured sparid species in southern Spain.
Regularized phase tomography enables study of mineralized and unmineralized tissue in porous bone scaffold
Authors: LANGER M. , LIU Y. , TORTELLI F. , CLOETENS P. , CANCEDDA R. , PEYRIN F.
Published: January 2010

Regularized phase tomography was used to image non‐calcified fibrous matrix in in vitro cell‐cultivated porous bone scaffold samples. 3D micro‐architecture of bone and bone scaffold has previously been studied by micro‐computed tomography, synchrotron radiation (SR) micro‐computed tomography and microdiffraction. However, neither of these techniques can resolve the low‐calcified immature pre‐bone fibrous structures. Skelite porous scaffold discs were seeded with osteoblasts, a combination of osteoblast and pre‐osteoclasts and, as controls, with pre‐osteoclasts only, and then cultivated for 8 weeks. They were subsequently imaged using SR propagation‐based phase contrast imaging. Reconstructions using a regularized holographic phase tomography approach were compared to standard (absorption) SR micro‐computed tomography, which show that quantitative analysis, such as volume and thickness measurements, of both the calcified fraction and the immature bone matrix in the reconstructed volumes is enabled. Indications of the effect of this type of culture on Skelite, such as change in mineralization and deposit of mature bone on the walls of the scaffold, are found. The results are verified with a histological study.
Phylogeny and taxonomy of the Prenolepis genus‐group of ants (Hymenoptera: Formicidae)
Authors: LaPOLLA JOHN S. , BRADY SEÁN G. , SHATTUCK STEVEN O.
Published: January 2010

Abstract. We investigated the phylogeny and taxonomy of the Prenolepis genus‐group, a clade of ants we define within the subfamily Formicinae comprising the genera Euprenolepis, Nylanderia, gen. rev., Paraparatrechina, gen. rev. & stat. nov., Paratrechina, Prenolepis and Pseudolasius. We inferred a phylogeny of the Prenolepis genus‐group using DNA sequence data from five genes (CAD, EF1αF1, EF1αF2, wingless and COI) sampled from 50 taxa. Based on the results of this phylogeny the taxonomy of the Prenolepis genus‐group was re‐examined. Paratrechina (broad sense) species segregated into three distinct, robust clades. Paratrechina longicornis represents a distinct lineage, a result consistent with morphological evidence; because this is the type species for the genus, Paratrechina is redefined as a monotypic genus. Two formerly synonymized subgenera, Nylanderia and Paraparatrechina, are raised to generic status in order to provide names for the other two clades. The majority of taxa formerly placed in Paratrechina, 133 species and subspecies, are transferred to Nylanderia, and 28 species and subspecies are transferred to Paraparatrechina. In addition, two species are transferred from Pseudolasius to Paraparatrechina and one species of Pseudolasius is transferred to Nylanderia. A morphological diagnosis for the worker caste of all six genera is provided, with a discussion of the morphological characters used to define each genus. Two genera, Prenolepis and Pseudolasius, were not recovered as monophyletic by the molecular data, and the implications of this result are discussed. A worker‐based key to the genera of the Prenolepis genus‐group is provided.
Phenotypic variability, neurological outcome and genetics background of 6‐pyruvoyl‐tetrahydropterin synthase deficiency
Authors: Leuzzi V , Carducci Ca , Carducci Cl , Pozzessere S , Burlina A , Cerone R , Concolino D , Donati MA , Fiori L , Meli C , Ponzone A , Porta F , Strisciuglio P , Antonozzi I , Blau N
Published: January 2010

Leuzzi V, Carducci Ca, Carducci Cl, Pozzessere S, Burlina A, Cerone R, Concolino D, Donati MA, Fiori L, Meli C, Ponzone A, Porta F, Strisciuglio P, Antonozzi I, Blau N. Phenotypic variability, neurological outcome and genetics background of 6‐pyruvoyl‐tetrahydropterin synthase deficiency.

This study aimed to investigate the clinical variability and factors implied in the outcome of 6‐pyruvoyl‐tetrahydropterin synthase deficiency (PTPSd). Biochemical and clinical phenotype, treatment variables, and 6‐pyruvoyl‐tetrahydropterin synthase (PTS) genotype, were explored retrospectively in 19 Italian patients (12 males and 7 females, aged 4 months to 33 years).

According to the level of biogenic amines in cerebrospinal fluid (CSF) at the diagnosis, the patients were classified as mild (6) (normal level) or severe (13) (abnormal low level) form (MF and SF, respectively). Blood Phe ranged from 151 to 1053 µmol/l in MF (mean ± SD: 698 ± 403) and 342–2120 µmol/l in SF (mean ± SD: 1175 ± 517) (p = 0.063). Patients with MF showed a normal neurological development (a transient dystonia was detected in one), while all SF patients except one presented with severe neurological impairment and only four had a normal neurological development. The outcome of the SF was influenced by the precocity of the treatment. Serial CSF examinations revealed a decline of 5‐hydroxyindolacetic acid in MFs and an incomplete restoration of neurotransmitters in SFs: neither obviously affected the prognosis. PTS gene analysis detected 17 different mutations (seven so far unreported) (only one affected allele was identified in three subjects). A good correlation was found between genotype and clinical and biochemical phenotype.

The occurrence of brain neurotransmitter deficiency and its early correction (by the therapy) are the main prognostic factors in PTPSd.
Cardiac conduction improvement in two heterozygotes for primary carnitine deficiency on l‐carnitine supplementation
Authors: Sarafoglou K , Tridgell AHC , Bentler K , Redlinger‐Grosse K , Berry SA , Schimmenti LA
Published: January 2010

Sarafoglou K, Tridgell AHC, Bentler K, Redlinger‐Grosse K, Berry SA, Schimmenti LA. Cardiac conduction improvement in two heterozygotes for primary carnitine deficiency on l‐carnitine supplementation.

Expanded newborn screening (NBS) for free carnitine levels has led to the identification of a larger number of heterozygous infants of undiagnosed mothers affected with systemic primary carnitine deficiency (PCD), which in turn leads to the identification of other undiagnosed heterozygous family members. There is an increasing recognition that individuals heterozygous for mutations of genes involved in fatty acid oxidation (FAO) may become symptomatic under environmental stress (fasting, prolonged exercise and illness). Considering the importance of carnitine in FAO, its role in heart and bowel function and in lipid metabolism, what is still little known is the phenotypic variability, biochemical parameters and clinical course of PCD heterozygotes with consistently low‐to‐normal levels to low levels of carnitine over a lifetime. We report on three generations of a family—an asymptomatic PCD heterozygous infant identified through NBS that led to the diagnosis of her asymptomatic PCD‐affected mother and the heterozygous status of the maternal grandparents who report some cardiac symptoms that overlap with PCD that improved with l‐carnitine supplementation.
Pink‐creamy whole blood in a 3‐month‐old infant with a homozygous deletion in the lipoprotein lipase gene
Authors: Avis HJ , Scheffer HJ , Kastelein JJP , Dallinga‐Thie GM , Wijburg FA
Published: January 2010

Avis HJ, Scheffer HJ, Kastelein JJP, Dallinga‐Thie GM, Wijburg FA. Pink‐creamy whole blood in a 3‐month‐old infant with a homozygous deletion in the lipoprotein lipase gene.
Biological activity of soils under rubber trees (Hevea brasiliensis) affected by trunk phloem necrosis
Authors: Wongcharoen A. , Pando A. , Nandris D. , Hanboonsoon Y. , Hartmann C. , Rouland‐Lefèvre C.
Published: January 2010

Trunk phloem necrosis (TPN) is currently one of the most economically important diseases of the rubber tree (Hevea brasiliensis). Investigations of the aetiology of the disorder have failed to identify any biotic causal agents but a multidisciplinary study has recently suggested that the disease may be caused by a combination of exogenous and endogenous stresses. These stresses can affect plant physiology and may also have an impact on soil biology and soil biochemistry. In this study, macrofauna diversity and soil enzyme activities were compared for the soils under trees affected by TPN and under healthy trees. Principal component analysis revealed associations between TPN and macrofauna diversity and soil enzyme activities. Groups of ants and termites were associated with the soil under healthy and trees affected by TPN respectively. Polysaccharidase (cellulase, xylanase and, particularly, amylase) activities were lower in the soil under trees affected by TPN whereas N‐acetylglucosaminidase activity was higher. These findings suggested that termite density and N‐acetylglucosaminidase activity are useful indicators of TPN, while lower values of polysaccharidases activities are the consequence of this disease. The impact of TPN on soil ecosystem was discussed.
Chronic kidney disease as a risk factor for Clostridium difficile infection
Authors: EDDI RODNEY , MALIK MOHAMMED N. , SHAKOV RADA , BADDOURA WALID J. , CHANDRAN CHANDRA , DEBARI VINCENT A.
Published: January 2010

Introduction:  Clostridium difficile‐associated diarrhoea (CDAD) is the most common cause of nosocomial diarrhoea in the USA. In this study, we sought to determine the association between chronic kidney disease (CKD) and CDAD.

Methods:  A case–control study was designed to determine the association between CKD and CDAD in an urban hospital. Over a 2‐year period, all patients diagnosed with CDAD (n = 188) were included as cases and the prevalence of CKD was calculated. Age‐ and sex‐matched patients without CDAD were considered as controls with a ratio of 2:1 controls to cases. The prevalence of different stages of advanced CKD (stages 3–5) was determined and compared between groups. Also the calculated odds ratios (OR) were adjusted for multiple possible confounding variables using logistic regression analysis.

Results:  There was no significant difference in prevalence of advanced CKD between cases and controls (OR = 1.38, 95% confidence intervals (CI) = 0.90–2.12, P = 0.1365). The association between CKD and CDAD remained insignificant in subjects with CKD stages 3–5 who were not on dialysis (OR = 1.07, 95% CI = 0.65–1.77), P = 0.7970). However, the group with end‐stage renal disease on dialysis showed a significant association (OR = 2.60, 95% CI = 1.25–5.41, P = 0.0165). Controlling for antibiotics as a possible confounding variable, yielded an OR that was not statistically significant (OR = 2.05, 95% CI = 0.94–4.47, P = 0.07), but still showing a trend towards increased risk.

Conclusion:  End‐stage renal disease may increase the risk of acquiring CDAD through unknown mechanisms. This suggests implementing better surveillance strategies for these patients and eliminating the known risk factors for CDAD.