bluuebunny/crossref_metadata_2025 huggingface dataset:
Commensal-dependent expression of IL-25 regulates the IL-23–IL-17 axis in the intestine
Authors: Zaph Colby , Du Yurong , Saenz Steven A. , Nair Meera G. , Perrigoue Jacqueline G. , Taylor Betsy C. , Troy Amy E. , Kobuley Dmytro E. , Kastelein Robert A. , Cua Daniel J. , Yu Yimin , Artis David
Published: September 2008

Alterations in the composition of intestinal commensal bacteria are associated with enhanced susceptibility to multiple inflammatory diseases, including those conditions associated with interleukin (IL)-17–producing CD4+ T helper (Th17) cells. However, the relationship between commensal bacteria and the expression of proinflammatory cytokines remains unclear. Using germ-free mice, we show that the frequency of Th17 cells in the large intestine is significantly elevated in the absence of commensal bacteria. Commensal-dependent expression of the IL-17 family member IL-25 (IL-17E) by intestinal epithelial cells limits the expansion of Th17 cells in the intestine by inhibiting expression of macrophage-derived IL-23. We propose that acquisition of, or alterations in, commensal bacteria influences intestinal immune homeostasis via direct regulation of the IL-25–IL-23–IL-17 axis.
Effects of magnetic field and target plasma on the penetration behaviour of compact toroid plasma by heat load measurements in CPD
Authors: Bhattacharyay R. , Zushi H. , Fukumoto N. , Nagata M. , Nishino N. , Honma H. , Kawakami K. , Yoshida N. , Kawasaki S. , Yoshinaga T. , Sasaki K. , Hasegawa M. , Sato K.N. , Sakamoto M. , Nakamura K. , Idei H. , Nakashima H. , Higashijima A.
Published: August 2008

Compact toroid (CT) injection experiments have recently been carried out in a compact plasma wall interaction experimental device (CPD) in the presence of toroidal as well as poloidal magnetic field and low density RF target plasma. Spectroscopic studies have been carried out using an IR camera, an IR spectrometer and an Hα monitor to estimate the heat load deposition on the target and also to study the magnetic field as well as RF target plasma effects on CT plasma penetration behaviour. Using IR camera imaging of the target plate, heat load deposition on it has been estimated from the observed temperature rise of the plate and has been compared with the energy content of CT plasma. The estimation suggests that nearly 700 J of energy is uniformly deposited on the target plate under vacuum condition without any magnetic field. On the other hand, post exposure analysis of target plate using scanning electron microscopy has indicated local melting of the target plate. It has been observed that radial penetration of CT towards the target plate is reduced with the increase in the external magnetic field, both toroidal and poloidal. Moreover, the presence of RF target plasma in external magnetic fields seems to change the CT penetration behaviour significantly.
Hurricane Katrina Deaths, Louisiana, 2005
Authors: Brunkard Joan , Namulanda Gonza , Ratard Raoult
Published: August 2008

Objective: Hurricane Katrina struck the US Gulf Coast on August 29, 2005, causing unprecedented damage to numerous communities in Louisiana and Mississippi. Our objectives were to verify, document, and characterize Katrina-related mortality in Louisiana and help identify strategies to reduce mortality in future disasters.

Methods: We assessed Hurricane Katrina mortality data sources received in 2007, including Louisiana and out-of-state death certificates for deaths occurring from August 27 to October 31, 2005, and the Disaster Mortuary Operational Response Team's confirmed victims' database. We calculated age-, race-, and sex-specific mortality rates for Orleans, St Bernard, and Jefferson Parishes, where 95% of Katrina victims resided and conducted stratified analyses by parish of residence to compare differences between observed proportions of victim demographic characteristics and expected values based on 2000 US Census data, using Pearson chi square and Fisher exact tests.

Results: We identified 971 Katrina-related deaths in Louisiana and 15 deaths among Katrina evacuees in other states. Drowning (40%), injury and trauma (25%), and heart conditions (11%) were the major causes of death among Louisiana victims. Forty-nine percent of victims were people 75 years old and older. Fifty-three percent of victims were men; 51% were black; and 42% were white. In Orleans Parish, the mortality rate among blacks was 1.7 to 4 times higher than that among whites for all people 18 years old and older. People 75 years old and older were significantly more likely to be storm victims (P < .0001).

Conclusions: Hurricane Katrina was the deadliest hurricane to strike the US Gulf Coast since 1928. Drowning was the major cause of death and people 75 years old and older were the most affected population cohort. Future disaster preparedness efforts must focus on evacuating and caring for vulnerable populations, including those in hospitals, long-term care facilities, and personal residences. Improving mortality reporting timeliness will enable response teams to provide appropriate interventions to these populations and to prepare and implement preventive measures before the next disaster. (Disaster Med Public Health Preparedness. 2008;2:215–223)
Regulation of muscle development by DPF3, a novel histone acetylation and methylation reader of the BAF chromatin remodeling complex
Authors: Lange Martin , Kaynak Bogac , Forster Ulrike B. , Tönjes Martje , Fischer Jenny J. , Grimm Christina , Schlesinger Jenny , Just Steffen , Dunkel Ilona , Krueger Tammo , Mebus Siegrun , Lehrach Hans , Lurz Rudi , Gobom Johan , Rottbauer Wolfgang , Abdelilah-Seyfried Salim , Sperling Silke
Published: September 2008

Chromatin remodeling and histone modifications facilitate access of transcription factors to DNA by promoting the unwinding and destabilization of histone–DNA interactions. We present DPF3, a new epigenetic key factor for heart and muscle development characterized by a double PHD finger. DPF3 is associated with the BAF chromatin remodeling complex and binds methylated and acetylated lysine residues of histone 3 and 4. Thus, DPF3 may represent the first plant homeodomains that bind acetylated lysines, a feature previously only shown for the bromodomain. During development Dpf3 is expressed in the heart and somites of mouse, chicken, and zebrafish. Morpholino knockdown of dpf3 in zebrafish leads to incomplete cardiac looping and severely reduced ventricular contractility, with disassembled muscular fibers caused by transcriptional deregulation of structural and regulatory proteins. Promoter analysis identified Dpf3 as a novel downstream target of Mef2a. Taken together, DPF3 adds a further layer of complexity to the BAF complex by representing a tissue-specific anchor between histone acetylations as well as methylations and chromatin remodeling. Furthermore, this shows that plant homeodomain proteins play a yet unexplored role in recruiting chromatin remodeling complexes to acetylated histones.
Shugoshin-2 is essential for the completion of meiosis but not for mitotic cell division in mice
Authors: Llano Elena , Gómez Rocío , Gutiérrez-Caballero Cristina , Herrán Yurema , Sánchez-Martín Manuel , Vázquez-Quiñones Luis , Hernández Teresa , de Álava Enrique , Cuadrado Ana , Barbero José Luis , Suja José A. , Pendás Alberto M.
Published: September 2008

Shugoshin-2 (SGOL2) is one of the two mammalian orthologs of the Shugoshin/Mei-S322 family of proteins that regulate sister chromatid cohesion by protecting the integrity of the multiprotein cohesin complexes. This protective system is essential for faithful chromosome segregation during mitosis and meiosis, which is the physical basis of Mendelian inheritance. Regardless of its evolutionary conservation from yeast to mammals, little is known about the in vivo relevance and specific role that SGOL2 plays in mammals. Here we show that disruption of the gene encoding mouse SGOL2 does not cause any alteration in sister chromatid cohesion in embryonic cultured fibroblasts and adult somatic tissues. Moreover, mutant mice develop normally and survive to adulthood without any apparent alteration. However, both male and female Sgol2-deficient mice are infertile. We demonstrate that SGOL2 is necessary for protecting centromeric cohesion during mammalian meiosis I. In vivo, the loss of SGOL2 promotes a premature release of the meiosis-specific REC8 cohesin complexes from anaphase I centromeres. This molecular alteration is manifested cytologically by the complete loss of centromere cohesion at metaphase II leading to single chromatids and physiologically with the formation of aneuploid gametes that give rise to infertility.
The RNA-binding protein XSeb4R: a positive regulator of VegT mRNA stability and translation that is required for germ layer formation in Xenopus
Authors: Souopgui Jacob , Rust, Barbara , Vanhomwegen Jessica , Heasman Janet , Henningfeld Kristine A. , Bellefroid Eric , Pieler Tomas
Published: September 2008

VegT represents a localized maternal determinant essentially required for endoderm formation in Xenopus. Here, we report on the identification of the RNA-binding protein XSeb4R as a positive regulator of VegT. XSeb4R interacts directly with the 3′-untranslated region of VegT mRNA, stabilizes it, and stimulates translation. Ablation of XSeb4R activity results in impairment of endoderm and mesoderm formation, while ectopic expression of XSeb4R in ectodermal cells induces endodermal and mesodermal gene expression. These observations unravel a novel mode of VegT regulation at the post-transcriptional level that is essential for germ layer formation in Xenopus.
eIF3a cooperates with sequences 5′ of uORF1 to promote resumption of scanning by post-termination ribosomes for reinitiation on GCN4 mRNA
Authors: Szamecz Béla , Rutkai Edit , Cuchalová Lucie , Munzarová Vanda , Herrmannová Anna , Nielsen Klaus H. , Burela Laxminarayana , Hinnebusch Alan G. , Valášek Leoš
Published: September 2008

Yeast initiation factor eIF3 (eukaryotic initiation factor 3) has been implicated in multiple steps of translation initiation. Previously, we showed that the N-terminal domain (NTD) of eIF3a interacts with the small ribosomal protein RPS0A located near the mRNA exit channel, where eIF3 is proposed to reside. Here, we demonstrate that a partial deletion of the RPS0A-binding domain of eIF3a impairs translation initiation and reduces binding of eIF3 and associated eIFs to native preinitiation complexes in vivo. Strikingly, it also severely blocks the induction of GCN4 translation that occurs via reinitiation. Detailed examination unveiled a novel reinitiation defect resulting from an inability of 40S ribosomes to resume scanning after terminating at the first upstream ORF (uORF1). Genetic analysis reveals a functional interaction between the eIF3a-NTD and sequences 5′ of uORF1 that is critically required to enhance reinitiation. We further demonstrate that these stimulatory sequences must be positioned precisely relative to the uORF1 stop codon and that reinitiation efficiency after uORF1 declines with its increasing length. Together, our results suggest that eIF3 is retained on ribosomes throughout uORF1 translation and, upon termination, interacts with its 5′ enhancer at the mRNA exit channel to stabilize mRNA association with post-termination 40S subunits and enable resumption of scanning for reinitiation downstream.
Preparation of Baskets for Drosophila Egg Collections, Treatments, and Incubations
Authors: Shermoen Antony W.
Published: September 2008

INTRODUCTION

The key consideration in preparation of baskets for Drosophila embryo collection is basket size, since the collected embryos must be in approximately one monolayer (or more). This article describes two baskets (homemade and commercial) used by the author for large numbers of embryos.
Subbing Slides
Authors: Paul Carol Ann , Beltz Barbara , Berger-Sweeney Joanne
Published: September 2008

INTRODUCTION

When mounting sections on a slide, it is essential that the slide be ultraclean. To ensure that the section sticks to the slide, slides can be covered with a thin layer of gelatin. The process of cleaning and gelatin-coating is called “subbing.” Several boxes of slides should be subbed at a time and stored, covered, in the refrigerator until they are needed. Subbed slides can also be purchased commercially.
Analyzing Glucose Phosphate Isomerase Isozymes in Chimeric Mouse Tissues by Electrophoresis
Authors: Nagy Andras , Gertsenstein Marina , Vintersten Kristina , Behringer Richard
Published: September 2008

INTRODUCTION

Mouse strains carry different alleles at the ubiquitously expressedGpi1(glucose phosphate isomerase) locus (Gpi1a,Gpi1b,Gpi1c), and this is the basis for a widely used method for determining the genotypic composition of different tissues in mouse chimeras. To determine chimerism by this method, it is necessary to separate the differently charged isozymes from tissue homogenates electrophoretically and to visualize them using a color reaction. Because GPI is a dimer, tissues that normally form by cell fusion (e.g., skeletal muscle) have a heterodimeric form of GPI in a chimera.
Phenotypic Analysis of Arabidopsis Mutants: Heavy Metal Stress
Authors: Lee David , Schroeder Julian
Published: September 2008

INTRODUCTION

Arsenic (As) is a major environmental contaminant, arising from both industrial and agricultural use, and is one of the metals given top priority for remediation by the U.S. Department of Energy on the basis of its toxicity and widespread contamination. However, many of the molecular and genetic mechanisms contributing to arsenic toxicity and detoxification remain unknown in plants. The toxicity of arsenic depends on the arsenic oxidation state. The main arsenic oxidation states are As(III) and As(V), both of which are highly toxic to plants and animals. In plants, As(III) disrupts the sulfhydryl groups of proteins and induces the production of hydrogen peroxide, which leads to membrane damage, whereas As(V) uncouples phosphorylation in mitochondria and at high levels can inhibit seed germination. As described in this protocol, resistance or sensitivity to arsenic can be easily scored by measuring germination and root bending in exposed seedlings.
Noninvasive Human Nuclear Transfer with Embryonic Stem Cells: Figure 1.
Authors: McElroy Sohyun L. , Reijo Pera Renee A.
Published: September 2008

INTRODUCTION

In somatic cell nuclear transfer (SCNT), the nucleus of a somatic cell is transferred to an enucleated oocyte for reprogramming to an embryonic cell state through the use of the endogenous machinery. SCNT technology has been used to produce offspring, establish embryonic stem cells, and study epigenetic reprogramming, as mediated by oocytes, in several animal species. In humans, there are ethical and practical issues that limit availability of oocytes donated by women of reproductive age specifically for research. Thus, there is a need to more exhaustively explore alternatives, including oocyte sources and different SCNT protocols. Nuclear transfer (NT) techniques are important factors that impact development of NT embryos. The procedures of enucleation of oocyte genetic material and introduction of the donor nucleus vary depending on species and laboratories. Hoechst staining has been used successfully for invasive enucleation in many animal studies, though it is known that Hoechst staining and ultraviolet (UV) light can damage oocyte mitochondrial DNA. More recently, noninvasive NT techniques that rely on polarized microscopic imaging systems have been used to visualize the meiotic spindle without DNA staining and UV illumination. This protocol describes a method for noninvasive human nuclear transfer by visualizing the oocyte spindle without DNA staining.
Preparation of Mouse Embryonic Fibroblast Feeder Cells for Human Embryonic Stem Cell Culture
Authors: McElroy Sohyun L. , Reijo Pera Renee A.
Published: September 2008

INTRODUCTION

Embryonic stem cells (ESCs) are derived from the inner cell mass of day 5-6 blastocysts. ESCs are pluripotent, meaning that they are able to differentiate into all derivatives of the three primary germ layers (ectoderm, endoderm, and mesoderm). In order to maintain the undifferentiated status of human ESCs (hESCs), feeder cells are used to provide both a suitable attachment substrate and critical soluble factors. Since the first hESC lines were established on mouse embryonic fibroblasts (MEFs), mitotically inactivated MEFs have commonly been used for supporting the culture of undifferentiated hESCs. Some previous studies suggest that MEFs may support hESC growth better than the human feeder cells typically isolated from post-natal tissues. This protocol describes a method for isolation and irradiation of MEFs for use in hESC culture.
Method for Single-Cell Sorting and Expansion of Genetically Modified Human Embryonic Stem Cells
Authors: Nicholas Cory R. , Reijo Pera Renee A.
Published: September 2008

INTRODUCTION

The potential for human embryonic stem cells (hESCs) to differentiate into all three embryonic germ layers, and the germline, can be used as a tool to understand the mechanisms of cell lineage-specific differentiation and development in vitro and in vivo. Therefore, it is important to develop technologies for the manipulation of hESCs and their effective utilization in research. One such manipulation is genetic modification. Reliable methods for genetic modification of hESCs with ubiquitous and tissue-specific reporters will assist in the identification and isolation of specific cell types following hESC differentiation. This protocol describes a method for hESC transduction with a ubiquitous eGFP reporter construct using self-inactivating lentivirus, followed by single-cell isolation and expansion of homogenous genetically modified hESCs using fluorescence-activated cell sorting (FACS).
DNA Methylation Analysis of Human Imprinted Loci by Bisulfite Genomic Sequencing
Authors: Angeles Vanessa T. , Reijo Pera Renee A.
Published: September 2008

INTRODUCTION

The genomic imprinting mechanism regulates differential expression of paternally and maternally derived genes. During development, primordial germ cells (PGCs) have a unique methylation pattern which undergoes demethylation, both globally and at imprinted loci. Later, the methylation status of imprinted genes is re-established during gametogenesis by de novo methylation. Errors in the methylation patterns of imprinted genes have been associated with syndromes such as Beckwith-Wiedemann, Angelman, and Prader-Willi in humans. Human embryonic stem cells (hESCs) have unique methylation patterns compared to somatic stem cells and cancer cells. The status of allelic expression or methylation of human imprinted genes can be studied in detail in human embryos/blastocysts and hESCs to understand early development and differentiation. This protocol describes the bisulfite genomic sequencing method for analysis of methylation patterns of imprinted loci in human embryonic stem cells. Multiple imprinted loci can be analyzed by performing the following protocol with gene-specific primers.
Metaphase Spreads and Spectral Karyotyping of Human Embryonic Stem Cells
Authors: Nguyen Ha Nam , Reijo Pera Renee A.
Published: September 2008

INTRODUCTION

Spectral karyotyping is a powerful technique for enumeration of chromosomes in plant, animal, and human cells with high precision. It is also used as a tool for detecting and diagnosing abnormalities in individual chromosomes, such as deletions, duplications, and translocations. There are three basic steps to obtaining good spectral karyotypes: (1) making metaphase spreads, (2) hybridizing probes to the DNA, and (3) analyzing the spectral characteristics of the chromosomes. There are two techniques for spectral karyotyping that are widely used: spectral karyotype (SKY) and multicolor fluorescence in situ hybridization (M-FISH). Both techniques use multicolored fluorescent probes which emit at different wavelengths (200-700 nm) upon light excitation to hybridize to the target DNA sequences. The methods differ in the way in which light is captured and analyzed by the corresponding software. In this protocol, methods for making metaphase spreads from human embryonic stem cells (hESCs) for spectral karyotyping (SKY) using the spectral imaging system and software from Applied Spectral Imaging are described.
Distinct difference of flaA genotypes of Legionella pneumophila between isolates from bath water and cooling tower water
Authors: Amemura‐Maekawa Junko , Kura Fumiaki , Chang Bin , Suzuki‐Hashimoto Atsuko , Ichinose Masayuki , Endo Takuro , Watanabe Haruo
Published: September 2008

To investigate the genetic difference of Legionella pneumophila in human‐made environments, we collected isolates of L. pneumophila from bath water (n = 167) and cooling tower water (n = 128) primarily in the Kanto region in 2001 and 2005. The environmental isolates were serogrouped and sequenced for a target region of flaA. A total of 14 types of flaA genotypes were found: 10 from cooling tower water and nine from bath water. The flaA genotypes of isolates from cooling tower water were quite different from those of bath water.
THIS ARTICLE HAS BEEN RETRACTED Anti‐adult T‐cell leukemia effects of a novel synthetic retinoid, Am80 (Tamibarotene)
Authors: Nakazato Tetsuro , Okudaira Taeko , Ishikawa Chie , Nakama Shinji , Sawada Shigeki , Tomita Mariko , Uchihara Jun‐nosuke , Taira Naoya , Masuda Masato , Tanaka Yuetsu , Ohshiro Kazuiku , Takasu Nobuyuki , Mori Naoki
Published: September 2008

Clinical trials for treatment of adult T‐cell leukemia (ATL) caused by human T‐cell leukemia virus type I (HTLV‐I) using all‐trans‐retinoic acid (ATRA) have shown satisfactory therapeutic responses, although efficacies were limited. Recently, many synthetic retinoids have been developed and among them, a novel synthetic retinoid, Am80 (Tamibarotene) is an RARα‐ and RARβ‐specific retinoid expected to overcome ATRA resistance. The present study examined the inhibitory effects of Am80 on HTLV‐I‐infected T‐cell lines and ATL cells. Am80 had negligible growth inhibition of peripheral blood mononuclear cells but marked growth inhibition of both HTLV‐I‐infected T‐cell lines and ATL cells. Am80 arrested cells in the G1 phase of the cell cycle and induced apoptosis in HTLV‐I‐infected T‐cell lines. It inhibited also the phosphorylation of IκBα and NF‐κB‐DNA binding, in conjunction with reduction of expression of proteins involved in the G1/S cell cycle transition and apoptosis. Am80 also inhibited the expression of JunD, resulting in suppression of AP‐1‐DNA binding. Furthermore, severe combined immunodeficient mice with tumors induced by subcutaneous inoculation of HTLV‐I‐infected T cells, responded to Am80 treatment with partial regression of tumors and no side‐effects. These findings demonstrate that Am80 is a potential inhibitor of NF‐κB and AP‐1, and is a potentially useful therapeutic agent against ATL. (Cancer Sci 2008; 99: 2286–2294)
Overexpression of carboxylesterase‐2 results in enhanced efficacy of topoisomerase I inhibitor, irinotecan (CPT‐11), for multiple myeloma
Authors: Yano Hiroki , Kayukawa Satoshi , Iida Shinsuke , Nakagawa Chiharu , Oguri Tetsuya , Sanda Takaomi , Ding Jianming , Mori Fumiko , Ito Asahi , Ri Masaki , Inagaki Atsushi , Kusumoto Shigeru , Ishida Takashi , Komatsu Hirokazu , Inagaki Hiroshi , Suzuki Atsushi , Ueda Ryuzo
Published: September 2008

Multiple myeloma (MM) remains an incurable disease and further development of novel agents is needed. Because constitutive expression of topoisomerase I (TopoI) in MM cells and the efficacy of SN‐38, an active metabolite of irinotecan (CPT‐11), have been reported, we investigated the therapeutic potential of CPT‐11. Of the eight MM cell lines analyzed, four showed 50% inhibitory concentration values of less than 2 µg/mL for CPT‐11 and less than 2 ng/mL for SN‐38. This efficacy was partly explained by the high expression level of human carboxylesterase‐2 (hCE‐2) in MM cells. Interestingly, high expression of hCE‐2 represented the nature of normal plasma cells, suggesting that hCE‐2 could efficiently generate SN‐38 within the plasma cells. As expected, higher sensitivity to CPT‐11 was observed in hCE‐2‐overexpressing U266 cells than mock U266 cells. On the other hand, the expression levels of hCE‐1, TopoI, UGT1A and ABCG2 did not seem to be associated with the sensitivity of MM cells to CPT‐11. In a murine xenograft model inoculated s.c. with RPMI8226 cells, administration of CPT‐11 alone significantly reduced the tumor volume. When a combination of CPT‐11 and bortezomib was administered, the subcutaneous tumors completely disappeared. Thus, clinical trials on CPT‐11 in patients with relapsed or refractory MM are warranted. (Cancer Sci 2008; 99: 2309–2314)
The influence of labour on the pharmacokinetics of intravenously administered amoxicillin in pregnant women
Authors: Muller Anouk E. , Dörr P. Joep , Mouton Johan W. , De Jongh Joost , Oostvogel Paul M. , Steegers Eric A. P. , Voskuyl Rob A. , Danhof Meindert
Published: August 2008

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Examples exist that pharmacokinetics of drugs in pregnant women can differ from that in non‐pregnant individuals.

• In pregnant women before the onset of labour, the pharmacokinetics of amoxicillin is similar to that in non‐pregnant individuals, but for women during labour this is unknown.

WHAT THIS STUDY ADDS

• Labour influences the pharmacokinetics of amoxicillin.

• During labour and even more in the immediate postpartum period, the peripheral volume of distribution was decreased compared with pregnant women before the onset of labour.

• The volume of distribution increases with an increasing amount of oedema.

AIMS

Many physiological changes take place during pregnancy and labour. These might change the pharmacokinetics of amoxicillin, necessitating adjustment of the dose for prevention of neonatal infections. We investigated the influence of labour on the pharmacokinetics of amoxicillin.

METHODS

Pregnant women before and during labour were recruited and treated with amoxicillin intravenously. A postpartum dose was offered. Blood samples were obtained and amoxicillin concentrations were determined using high‐pressure liquid chromatography. The pharmacokinetics were characterized by nonlinear mixed‐effects modelling using NONMEM.

RESULTS

The pharmacokinetics of amoxicillin in 34 patients was best described by a three‐compartment model. Moderate interindividual variability was identified in CL, central and peripheral volumes of distribution. The volume of distribution (V) increased with an increasing amount of oedema. Labour influenced the parameter estimate of peripheral volume of distribution (V2). V2 was decreased during labour, and even more in the immediate postpartum period. For all patients the population estimates (mean ± SE) for CL and V were 21.1 ± 4.1 l h−1 (CL), 8.7 ± 6.6 l (V1), 11.8 ± 7.7 l (V2) and 20.5 ± 15.4 l (V3) respectively.

CONCLUSIONS

The peripheral distribution volume of amoxicillin in pregnant women during labour and immediately postpartum is decreased. However, these changes are not clinically relevant and do not warrant deviations from the recommended dosing regimen for amoxicillin during labour in healthy pregnant patients.
New treatments in type 2 diabetes: a focus on the incretin‐based therapies
Authors: Barnett Anthony H.
Published: September 2008

The demonstration that the incretin hormone glucagon‐like peptide 1 can improve glycaemic control in patients with type 2 diabetes has led to the rapid development during the last decade of promising new classes of agent for the management of type 2 diabetes. These agents possess a range of physiological effects that are associated with improved glycaemic control in diabetes including stimulation of glucose‐dependent insulin secretion, suppression of glucagon secretion, slowing of gastric emptying, and reduction of food intake. In addition, preclinical studies suggest that incretin‐based therapies may improve β‐cell function via enhancement of β‐cell mass and induction of genes important for differentiated β‐cell function. Exenatide, and the dipeptidyl peptidase‐4 inhibitors, sitagliptin and vildagliptin are already approved, and liraglutide is currently completing Phase 3 trials. As these agents and standard oral therapies for type 2 diabetes lower glucose levels through different, but potentially complementary mechanisms, their use in combination should provide effective, potentially additive, glycaemic control. The incretin‐based therapies also offer other advantages such as weight loss with exenatide and liraglutide, a reduced risk of hypoglycaemia, and as suggested by preclinical studies, a potential β‐cell preserving effect. Long‐term outcome and safety data are not available for these agents, but they appear generally well‐tolerated in comparison with existing therapies for type 2 diabetes. The multiple underlying glucose‐lowering actions of the incretin‐based therapies, as well as a lack of weight gain or even weight loss, make these important new additions to available antidiabetic agents expanding the treatment options available for patients.
Adjustment of L‐T4 substitutive therapy in pregnant women with subclinical, overt or post‐ablative hypothyroidism
Authors: Verga Uberta , Bergamaschi Silvia , Cortelazzi Donatella , Ronzoni Stefania , Marconi Anna Maria , Beck‐Peccoz Paolo
Published: September 2008

Objective  Maternal hyperthyrotropinaemia is associated with an increased risk of adverse maternal and neonatal outcomes. Physiological changes during pregnancy require an increased production of thyroid hormones (or an increase in daily substitutive doses of L‐T4 in hypothyroid patients) to meet the maternal and foetal needs. The aim of the study was to evaluate variations of substitutive L‐T4 doses that are able to maintain serum TSH between 0·5 and 2·5 mU/l in pregnant women with subclinical‐ (SH), overt‐ (OH) and post‐ablative (PH) hypothyroidism.

Design  This was a retrospective study on hypothyroid pregnant women referred to the out‐patient department between January 2004 and December 2006.

Patients and measurements  A total of 185 pregnant women were studied during gestation; 155 patients (76 SH, 52 OH, 27 PH) were already on L‐T4 before conception and 30 (SH) started L‐T4 therapy during gestation. Thyroid function and body weight were evaluated every 4–6 weeks.

Results  In the group of patients already treated before conception, 134 (86·5%) increased L‐T4 doses during gestation one or more times, eight (6%) reached a definitive therapeutic dosage within the 12th week of pregnancy, 64 (47·8%) within the 20th week and 62 (46·2%) within the 31st week. This initial L‐T4 increase at the first evaluation during pregnancy was 22·9 ± 9·8 µg/day. The final L‐T4 doses were significantly different depending on the aetiology, being 101·0 ± 24·6 µg/day in SH, 136·8 ± 30·4 µg/day in OH and 159·0 ± 24·6 µg/day in PH. The per cent increase of L‐T4, expressed as Δ% of absolute dose, was +70% in SH, +45% in OH and +49% in PH as compared to baseline dose. In SH patients diagnosed during gestation, the starting L‐T4 dose was higher than L‐T4 dose before pregnancy of SH patients already treated (75·4 ± 14·5 and 63·2 ± 20·1 µg/day, respectively), whereas the final doses were similar. L‐T4 dose was increased one or more times in 24 patients (80%), 8 reached the definitive dosage within the second trimester (33·3%) and 16 within the third trimester (66·7%).

Conclusions  Serum TSH and FT4 measurements are mandatory in pregnant patients and the optimal timing for increasing L‐T4 is the first trimester of pregnancy, though many patients require adjustments also during the second and third trimester. The aetiology of hypothyroidism influences the adjustment of L‐T4 therapy and SH patients needed a larger increase than OH and PH. Close monitoring during pregnancy appears to be mandatory in hypothyroid women.
Fission yeast chromatin assembly factor 1 assists in the replication‐coupled maintenance of heterochromatin
Authors: Dohke Kohei , Miyazaki Shota , Tanaka Katsunori , Urano Takeshi , Grewal Shiv I. S. , Murakami Yota
Published: August 2008

Chromatin assembly factor‐1 (CAF1) is a well‐conserved histone chaperone that loads the histone H3‐H4 complex onto newly synthesized DNA in vitro through interaction with the replication factor PCNA. CAF1 is considered to be involved in heterochromatin maintenance in several organisms, but the evidence is circumstantial and functional details have not been established. We identified fission yeast CAF‐1 (spCAF1), which interacts with PCNA in S phase. Depletion of spCAF1 caused defects in silencing at centromeric and mating locus heterochromatin, accompanied with a decrease in Swi6, the fission yeast HP1 homologue. Loss of spCAF1 destabilized both the silent and active states of chromatin at the meta‐stable heterochromatic region, with a more pronounced effect on the silent state, indicating that spCAF1 is involved in the maintenance of heterochromatin. Swi6 dissociated from heterochromatin during G1/S phase appears to associate with spCAF1. In early S phase, spCAF1 localized to replicating heterochromatin as well as euchromatin and remained associated with Swi6, and Swi6 then bound to heterochromatin. Taken together, we propose that spCAF1 functions in heterochromatin maintenance by recruiting dislocated Swi6 during replication to replicated heterochromatin at the replication fork.
Haemate® P/Humate‐P® for the treatment of von Willebrand disease: considerations for use and clinical experience
Authors: AUERSWALD G. , KREUZ W.
Published: August 2008

Summary.  von Willebrand disease (VWD) is a heterogeneous bleeding disorder with symptoms in affected patients ranging from mild effects to potentially devastating haemorrhagic events. Desmopressin (DDAVP) and von Willebrand factor/factor VIII (VWF/FVIII) concentrates are the principal treatments. Haemate® P/Humate‐P® is an intermediate‐purity VWF/FVIII concentrate with extensive clinical experience in VWD. This concentrate has been shown to correct haemostatic defects of VWD, with efficacy ratings of good/excellent in nearly all patients treated for bleeding or surgical events. Haemate P/Humate‐P has a high content of the high molecular weight (HMW) VWF multimer fraction, which has been shown to be very effective in achieving haemostasis. The HMW VWF multimer pattern in Haemate P/Humate‐P is more similar to that of normal human plasma (94% for Haemate P/Humate‐P vs. 100% for normal human plasma) than that of other VWF/FVIII concentrates and correlates with functional VWF activities including ristocetin cofactor activity (VWF:RCo) and collagen‐binding activity. The recommended dosing of Haemate P/Humate‐P is based preferentially on VWF:RCo activity, which is approximately twice that of FVIII:C (2.4:1). Haemate P/Humate‐P has been shown to be safe; no serious adverse events or cases of thrombosis have been observed in clinical trials and no documented cases of viral transmission in nearly three decades of clinical use. While DDAVP is effective in a large proportion of VWD patients, it may not provide adequate haemostasis in all situations. In such cases, Haemate P/Humate‐P is an effective replacement concentrate for all types of VWD in both adult and paediatric patients.
Identification of the most active interleukin‐32 isoform
Authors: Choi Ji‐Da , Bae Su‐Young , Hong Jae‐Woo , Azam Tania , Dinarello Charles A. , Her Erk , Choi Whan‐Soo , Kim Bo‐Kyung , Lee Chang‐Kwon , Yoon Do‐Young , Kim Sun‐Jong , Kim Soo‐Hyun
Published: September 2008

Cytokines are crucial in host defence against pathogens such as bacteria, viruses, fungi and parasites. A newly described cytokine, interleukin‐32 (IL‐32), induces various proinflammatory cytokines (tumour necrosis factor‐α, IL‐1β, IL‐6) and chemokines in both human and mouse cells through the nuclear factor‐κB and p38 mitogen‐activated protein kinase inflammatory signal pathway. The IL‐32 primarily acts on monocytic cells rather than T cells. In an attempt to isolate the IL‐32 soluble receptor, we used an IL‐32 ligand‐affinity column to purify neutrophil proteinase 3, which is a serine proteinase involved in many inflammatory diseases. IL‐32 has biological activity associated with Mycobacterium tuberculosis and chronic proinflammatory diseases such as rheumatoid arthritis. IL‐32 is transcribed as six alternative splice variants and the biological activity of each individual isoform remains unknown. Here, we cloned the complementary DNA of the four IL‐32 isoforms (α, β, γ and δ) that are the most representative IL‐32 transcripts. To produce recombinant protein with a high yield, the amino acids of two cysteine residues were mutated to serine residues, because serine residues are not conserved among different species. The multi‐step purified recombinant IL‐32 isoform proteins were assessed for their biological activities with different cytokine assays. The γ isoform of IL‐32 was the most active, although all isoforms were biologically active. The present study will provide a specific target to neutralize endogenous IL‐32, which may contribute to basic and clinical immunology.
Risk profiles for non‐adherence to antipsychotic medications
Authors: MCCANN T. V. , BOARDMAN G. , CLARK E. , LU S.
Published: September 2008

Poor adherence to medications is common in individuals with schizophrenia, and can lead to relapse and re‐hospitalization. This paper presents the findings of an Australian study of the factors affecting antipsychotic medication taking in individuals with schizophrenia. The Factors Influencing Neuroleptic Medication Taking Scale was used with a non‐probability sample of mental health service users. Ethics approval was obtained from a university and a hospital ethics committee. Data were analysed using spss version 15. Most participants had insight into their illness and were aware of the stigma of mental illness. Around 70% experienced annoying side effects, while nearly half admitted alcohol consumption. About one‐fifth admitted they had missed taking medications during the previous week. Significant others played a variable role in medication taking. Over 80% were satisfied with their relationships with health professionals, but were less satisfied with access to these professionals, especially psychiatrists. Logistic regression analysis showed that age, impact of medication side effects, and access to psychiatrists were independent predictors of medication omission. It is argued that medication taking is a complex issue, which needs to be taken into consideration in health professional training and measures to promote adherence.
User involvement in mental health care: the role of nurses. A literature review
Authors: STRINGER B. , VAN MEIJEL B. , DE VREE W. , VAN DER BIJL J.
Published: September 2008

This literature review was carried out to examine the effects of user involvement in shared decision‐making processes and the methods/tools available to psychiatric nurses to measure and encourage user involvement. A systematic literature review was then used in this study. Many studies indicate that an increased involvement of service users leads to better care, better treatment compliance, improved health outcomes and higher levels of patient satisfaction. The tools and methods described are designed to measure the ability to participate, the process of implementation and the evaluation of healthcare services. An adequate instrument to measure user involvement will be necessary to underpin the positive effects. Although care providers have a statutory duty to help shape user involvement, and the tools required are available, care providers are still insufficiently inclined to take up this duty.
The collaborative care practice model in the long‐term care of individuals with bipolar disorder: a case study
Authors: DAVIES M. A. , MCBRIDE L. , SAJATOVIC M.
Published: September 2008

Effective therapies for the long‐term care of individuals with bipolar disorder feature medication management and an interactional component between patients and care providers. In this paper, we discuss the effectiveness of applying a collaborative practice model to the long‐term care of individuals with bipolar disorder who receive outpatient treatment. The model emphasizes that patients are co‐managers of their illness and successful outcomes are enhanced within a supportive social environment. Key elements include joint patient–provider treatment planning and decision making. Several controlled trials have demonstrated that when the model is adopted, significant improvements in clinical and functional outcomes can be achieved. The paper concludes with a case study, which illustrates that the model can be successfully implemented in a clinical setting. Given that the clinical and research evidence demonstrates the effectiveness of the model, we suggest that advanced practice psychiatric nurses apply key elements of the collaborative practice model in their work with clients with bipolar disorder.
Evidence for multiple introductions of Centaurea stoebe micranthos (spotted knapweed, Asteraceae) to North America
Authors: MARRS R. A. , SFORZA R. , HUFBAUER R. A.
Published: September 2008

Invasive species’ success may depend strongly on the genetic resources they maintain through the invasion process. We ask how many introductions have occurred in the North American weed Centaurea stoebe micranthos (Asteraceae), and explore whether genetic diversity and population structure have changed as a result of introduction. We surveyed individuals from 15 European native range sites and 11 North American introduced range sites at six polymorphic microsatellite loci. No significant difference existed in the total number of alleles or in the number of private alleles found in each range. Shannon–Weaver diversity of phenotype frequencies was also not significantly different between the ranges, while expected heterozygosity was significantly higher in the invasive range. Population structure was similar between the native range and the invasive range, and isolation by distance was not significant in either range. Traditional assignment methods did not allocate any North American individuals to the sampled European populations, while Bayesian assignment methods grouped individuals into nine genetic clusters, with three of them shared between North America and Europe. Invasive individuals tended to have genetically admixed profiles, while natives tended to assign more strongly to a single cluster. Many North American individuals share assignment with Romania and Bulgaria, suggesting two separate invasions that have undergone gene flow in North America. Samples from three other invasive range sites were genetically distinct, possibly representing three other unique introductions. Multiple introductions and the maintenance of high genetic diversity through the introduction process may be partially responsible for the invasive success of C. stoebe micranthos.
Microbial communities and interactions in the lone star tick, Amblyomma americanum
Authors: CLAY K. , KLYACHKO O. , GRINDLE N. , CIVITELLO D. , OLESKE D. , FUQUA C.
Published: September 2008

To quantify microbial composition and interactions, we identified prokaryotic communities in the lone star tick (Amblyomma americanum) based on 16S rRNA gene sequences and direct probing. The lone star tick is the vector of emerging diseases and host to additional symbionts of unknown activity, and is representative of other blood‐sucking arthropods. We evaluated the potential for vertical (transovarial) transmission by molecular analysis of microbial symbionts from egg and larval clutches. Direct probing of adults (N = 8 populations from the southeastern and midwestern USA, 900 ticks total) revealed three vertically transmitted symbionts: a Coxiella symbiont occurred at 100% frequency, Rickettsia species occurred in 45–61% of all ticks in every population and an Arsenophonus symbiont occurred in 0–90% of ticks per population. Arsenophonus and Rickettsia exhibited significant heterogeneity in frequency among populations. The human pathogens Ehrlichia chafeensis and Borrelia lonestari were rare in most populations. Additional microbes were detected sporadically. Most ticks (78%) were co‐infected by two or three microbes but statistical analysis indicated no significant deviation from random co‐occurrence. Our findings indicate that microbial communities within lone star ticks are diverse, and suggest that direct probing for a wider range of prokaryotes and application of quantitative polymerase chain reaction (PCR) may provide further insights into microbial interactions within disease vectors. Our results also emphasize the close phylogenetic relationship between tick symbionts and human pathogens, and consistent differences in their prevalence.
Overproduction of Escherichia coli DNA polymerase DinB (Pol IV) inhibits replication fork progression and is lethal
Authors: Uchida Kaori , Furukohri Asako , Shinozaki Yutaka , Mori Tetsuya , Ogawara Daichi , Kanaya Shigehiko , Nohmi Takehiko , Maki Hisaji , Akiyama Masahiro
Published: August 2008

Escherichia coli dinB encodes the specialized DNA polymerase DinB (Pol IV), which is induced as part of the SOS stress‐response system and functions in translesion synthesis (TLS) to relieve the replicative Pol III that is stalled at DNA lesions. As the number of DinB molecules, even in unstressed cells, is greater than that required to accomplish TLS, it is thought that dinB plays some additional physiological role. Here, we overexpressed dinB under the tightly regulable arabinose promoter and looked for a distinct phenotype. Upon induction of dinB expression, progression of the replication fork was immediately inhibited at random genomic positions, and the colony‐forming ability of the cells was reduced. Overexpression of mutated dinB alleles revealed that the structural requirements for these two inhibitory effects and for TLS were distinct. The extent of in vivo inhibition displayed by a mutant DinB matched the extent of its in vitro impedance, at near‐physiological concentration, of a moving Pol III. We suggest that DinB targets Pol III, thereby acting as a brake on replication fork progression. Because the brake operates when cells have excess DinB, as they do under stress conditions, it may serve as a checkpoint that modulates replication to safeguard genome stability.
Cytolocalization of the PhoP response regulator in Salmonella enterica: modulation by extracellular Mg2+ and by the SCV environment
Authors: Sciara Mariela I. , Spagnuolo Carla , Jares‐Erijman Elizabeth , García Véscovi Eleonora
Published: August 2008

The PhoP/PhoQ two‐component system plays an essential role regulating numerous virulence phenotypes in Salmonella enterica. Previous work showed that PhoQ, the sensor protein, switches between the kinase‐ and the phosphatase‐dominant state in response to environmental Mg2+ availability. This switch defines the PhoP phosphorylation status and, as a result, the transcriptional activity of this regulator. In this work, using the FlAsH labelling technique, we examine PhoP cytolocalization in response to extracellular Mg2+ limitation in vitro and to the Salmonella‐containing vacuole (SCV) environment in macrophage cells. We demonstrate that in these PhoP/PhoQ‐inducing environments PhoP displays preferential localization to one cell pole, while being homogeneously distributed in the bacterial cytoplasm in repressing conditions. Polar localization is lost in the absence of PhoQ or when a non‐phosphorylatable PhoPD52A mutant is expressed. However, when PhoP transcriptional activation is achieved in a Mg2+‐ and PhoQ‐independent manner, PhoP regains asymmetric polar localization. In addition, we show that, in the analysed conditions, PhoQ cellular distribution does not parallel PhoP location pattern. These findings reveal that PhoP cellular location is dynamic and conditioned by its environmentally defined transcriptional status, showing a new insight in the PhoP/PhoQ system mechanism.