bluuebunny/crossref_metadata_2025 huggingface dataset:
Optogenetic control of epileptiform activity
Authors: Tønnesen Jan , Sørensen Andreas T. , Deisseroth Karl , Lundberg Cecilia , Kokaia Merab
Published: July 2009

The optogenetic approach to gain control over neuronal excitability both in vitro and in vivo has emerged as a fascinating scientific tool to explore neuronal networks, but it also opens possibilities for developing novel treatment strategies for neurologic conditions. We have explored whether such an optogenetic approach using the light-driven halorhodopsin chloride pump from
Natronomonas pharaonis
(NpHR), modified for mammalian CNS expression to hyperpolarize central neurons, may inhibit excessive hyperexcitability and epileptiform activity. We show that a lentiviral vector containing the NpHR gene under the calcium/calmodulin-dependent protein kinase IIα promoter transduces principal cells of the hippocampus and cortex and hyperpolarizes these cells, preventing generation of action potentials and epileptiform activity during optical stimulation. This study proves a principle, that selective hyperpolarization of principal cortical neurons by NpHR is sufficient to curtail paroxysmal activity in transduced neurons and can inhibit stimulation train-induced bursting in hippocampal organotypic slice cultures, which represents a model tissue of pharmacoresistant epilepsy. This study demonstrates that the optogenetic approach may prove useful for controlling epileptiform activity and opens a future perspective to develop it into a strategy to treat epilepsy.
A single modular serine protease integrates signals from pattern-recognition receptors upstream of the
Drosophila
Toll pathway
Authors: Buchon Nicolas , Poidevin Mickael , Kwon Hyun-Mi , Guillou Aurélien , Sottas Valentin , Lee Bok-Luel , Lemaitre Bruno
Published: July 2009

The
Drosophila
Toll receptor does not interact directly with microbial determinants, but is instead activated by a cleaved form of the cytokine-like molecule Spätzle. During the immune response, Spätzle is processed by complex cascades of serine proteases, which are activated by secreted pattern-recognition receptors. Here, we demonstrate the essential role of ModSP, a modular serine protease, in the activation of the Toll pathway by Gram-positive bacteria and fungi. Our analysis shows that ModSP integrates signals originating from the circulating recognition molecules GNBP3 and PGRP-SA and connects them to the Grass-SPE-Spätzle extracellular pathway upstream of the Toll receptor. It also reveals the conserved role of modular serine proteases in the activation of insect immune reactions.
Global discovery of primate-specific genes in the human genome
Authors: Tay Sen-Kwan , Blythe Jason , Lipovich Leonard
Published: July 2009

The genomic basis of primate phenotypic uniqueness remains obscure, despite increasing genome and transcriptome sequence data availability. Although factors such as segmental duplications and positive selection have received much attention as potential drivers of primate phenotypes, single-copy primate-specific genes are poorly characterized. To discover such genes genomewide, we screened a catalog of 38,037 human transcriptional units (TUs), compiled from EST and cDNA sequences in conjunction with the FANTOM3 transcriptome project. We identified 131 TUs from transcribed sequences residing within primate-specific insertions in 9-species sequence alignments and outside of segmental duplications. Exons of 120 (92%) of the TUs contained interspersed repeats, indicating that repeat insertions may have contributed to primate-specific gene genesis. Fifty-nine (46%) primate-specific TUs may encode proteins. Although primate-specific TU transcript lengths were comparable to known human gene mRNA lengths overall, 92 (70%) primate-specific TUs were single-exon. Thirty-two (24%) primate-specific TUs were localized to subtelomeric and pericentromeric regions. Forty (31%) of the TUs were nested in introns of known genes, indicating that primate-specific TUs may arise within older, protein-coding regions. Primate-specific TUs were preferentially expressed in reproductive organs and tissues (
P
< 0.011), consistent with the expectation that emergence of new, lineage-specific genes may accompany speciation or reproduction. Of the 33 primate-specific TUs with human Affymetrix microarray probe support, 21 were differentially expressed in human teratozoospermia. In addition to elucidating the likely functional relevance of primate-specific TUs to reproduction, we present a set of primate-specific genes for future functional studies, and we implicate nonduplicated pericentromeric and subtelomeric regions in gene genesis.
Expression of immediate-early genes reveals functional compartments within ocular dominance columns after brief monocular inactivation
Authors: Takahata Toru , Higo Noriyuki , Kaas Jon H. , Yamamori Tetsuo
Published: July 2009

Visual inputs from the 2 eyes in most primates activate alternating bands of cortex in layer 4C of primary visual cortex, thereby forming the well-studied ocular dominance columns (ODCs). In addition, the enzymatic reactivity of cytochrome oxidase (CO) reveals “blob” structures within the supragranular layers of ODCs. Here, we present evidence for compartments within ODCs that have not been clearly defined previously. These compartments are revealed by the activity-dependent mRNA expression of immediate-early genes (IEGs),
zif268
and
c-fos
, after brief periods of monocular inactivation (MI). After a 1–3-h period of MI produced by an injection of tetrodotoxin, IEGs were expressed in a patchy pattern that included infragranular layers, as well as supragranular layers, where they corresponded to the CO blobs. In addition, the expressions of IEGs in layer 4C were especially high in narrow zones along boundaries of ODCs, referred to here as the “border strips” of the ODCs. After longer periods of MI (>5 h), the border strips were no longer apparent. When either eyelid was sutured, changes in IEG expression were not evident in layer 4C; however, the patchy pattern of the expression in the infragranular and supragranular layers remained. These changes of IEG expression after MI indicate that cortical circuits involving the CO blobs of the supragranular layers include aligned groups of neurons in the infragranular layers and that the border strip neurons of layer 4C are highly active for a 3-h period after MI.
Substance Misuse and Early Psychosis
Authors: Tucker Peter
Published: July 2009

Objective: This paper sought to review current knowledge about the relationship between substance misuse and early psychosis.

Methods: A literature search was conducted using Medline and restricting the search to articles after 1996. Additional articles were sourced from reference lists of relevant articles.

Results: There is a high prevalence of substance misuse among persons with early psychosis, with cannabis and alcohol featuring prominently. Substance misuse is associated with earlier onset and possibly more positive symptoms, although apparently not with greater cognitive impairment. Cannabis appears to confer an increased likelihood of developing schizophrenia in biologically vulnerable individuals. Amphetamines also cause psychosis which may become chronic, although specific vulnerability to this effect is less well established. Many cases of so-called ‘drug-induced psychosis’ become diagnosed as schizophrenia in later years. Specific intervention programs report positive outcomes with regard to substance misuse and the course of psychosis.

Conclusions: Substance misuse should always be assessed in this patient group, bearing in mind the potential interactive causes of psychopathology. Intervention is of value in improving outcomes.
Patients who do Well and who do Less Well in an Inpatient Adolescent Unit
Authors: Mathai John , Bourne Angela
Published: July 2009

Objective: The primary aim of the study was to investigate whether patient characteristics such as age, sex, length of inpatient stay and reason for admission were related to positive or negative treatment outcome at an adolescent inpatient unit.

Method: This study employed a prospective cohort design in the form of a clinical audit. Health of the Nation Outcome Scales for Children and Adolescents (HoNOSCA) pre- and post-admission scores were compared for the whole sample and across diagnoses.

Results: While there was a significant reduction overall in symptoms following an inpatient stay, patient characteristics and reason for admission were not related to outcomes at a statistically significant level.

Conclusions: This inpatient unit generally functions as a short stay unit and as a result it is difficult to draw any meaningful conclusions as to what sort of patient would benefit most from an admission to the unit.
Soaring on the Wings of the Wind: Freud, Jews and Judaism
Authors: Kaplan Robert
Published: July 2009

Objectives: This paper looks at Freud's Jewish identity in the context of the Jewish experience in Eastern and Central Europe after 1800, using his family history and significant figures in his life as illustration. Sigmund Freud's life as a Jew is deeply paradoxical, if not enigmatic. He mixed almost exclusively with Jews while living all his life in an anti-Semitic environment. Yet he eschewed Jewish ritual, referred to himself as a godless Jew and sought to make his movement acceptable to gentiles. At the end of his life, dismayed by the rising forces of nationalism, he accepted that he was in his heart a Jew “in spite of all efforts to be unprejudiced and impartial”. The 18th century Haskalla (Jewish Enlightenment) was a form of rebellion against conformity and a means of escape from shtetl life. In this intense, entirely inward means of intellectual escape and revolt against authority, strongly tinged with sexual morality, we see the same tensions that were to manifest in the publication by a middle-aged Viennese neurologist of a truly revolutionary book to herald the new 20th century: The Interpretation of Dreams. Freud's life and work needs to be understood in the context of fin-de-siécle Vienna. Mitteleuropa, the cultural renaissance of Central Europe, resulted from the emancipation and urbanization of the burgeoning Jewish middle class, who adopted to the cosmopolitan environment more successfully than any other group. In this there is an extreme paradox: the Jewish success in Vienna was a tragedy of success.

Conclusions: Freud, despite a deliberate attempt to play down his Jewish origins to deflect anti-Semitic attacks, is the most representative Jew of his time and his thinking and work represents the finest manifestation of the Litvak mentality.
NF-κB activity marks cells engaged in receptor editing
Authors: Cadera Emily J. , Wan Fengyi , Amin Rupesh H. , Nolla Hector , Lenardo Michael J. , Schlissel Mark S.
Published: July 2009

Because of the extreme diversity in immunoglobulin genes, tolerance mechanisms are necessary to ensure that B cells do not respond to self-antigens. One such tolerance mechanism is called receptor editing. If the B cell receptor (BCR) on an immature B cell recognizes self-antigen, it is down-regulated from the cell surface, and light chain gene rearrangement continues in an attempt to edit the autoreactive specificity. Analysis of a heterozygous mutant mouse in which the NF-κB–dependent IκBα gene was replaced with a lacZ (β-gal) reporter complementary DNA (cDNA; IκBα+/lacZ) suggests a potential role for NF-κB in receptor editing. Sorted β-gal+ pre–B cells showed increased levels of various markers of receptor editing. In IκBα+/lacZ reporter mice expressing either innocuous or self-specific knocked in BCRs, β-gal was preferentially expressed in pre–B cells from the mice with self-specific BCRs. Retroviral-mediated expression of a cDNA encoding an IκBα superrepressor in primary bone marrow cultures resulted in diminished germline κ and rearranged λ transcripts but similar levels of RAG expression as compared with controls. We found that IRF4 transcripts were up-regulated in β-gal+ pre–B cells. Because IRF4 is a target of NF-κB and is required for receptor editing, we suggest that NF-κB could be acting through IRF4 to regulate receptor editing.
Mixture Model to Assess the Extent of Cross-Transmission of Multidrug-Resistant Pathogens in Hospitals
Authors: Mikolajczyk Rafael T. , Kauermann Göran , Sagel Ulrich , Kretzschmar Mirjam
Published: July 2009

Objective.

Creation of a mixture model based on Poisson processes for assessment of the extent of cross-transmission of multidrug-resistant pathogens in the hospital.

Methods.

We propose a 2-component mixture of Poisson processes to describe the time series of detected cases of colonization. The first component describes the admission process of patients with colonization, and the second describes the cross-transmission. The data set used to illustrate the method consists of the routinely collected records for methicillin-resistantStaphylococcus aureus(MRSA), imipenem-resistantPseudomonas aeruginosa, and multidrug-resistantAcinetobacter baumanniiover a period of 3 years in a German tertiary care hospital.

Results.

For MRSA and multidrug-resistantA. baumannii, cross-transmission was estimated to be responsible for more than 80% of cases; for imipenem-resistantP. aeruginosa, cross-transmission was estimated to be responsible for 59% of cases. For new cases observed within a window of less than 28 days for MRSA and multidrug-resistantA. baumanniior 40 days for imipenem-resistantP. aeruginosa, there was a 50% or greater probability that the cause was cross-transmission.

Conclusions.

The proposed method offers a solution to assessing of the extent of cross-transmission, which can be of clinical use. The method can be applied using freely available software (the package FlexMix in R) and it requires relatively little data.
Two-dimensional full particle simulation of the flow patterns in the scrape-off-layer plasma for upper- and lower-null point divertor configurations in tokamaks
Authors: Takizuka T. , Shimizu K. , Hayashi N. , Hosokawa M. , Yagi M.
Published: July 2009

The plasma flow in the scrape-off-layer (SOL) plays an important role in particle control in magnetic fusion reactors. The flow is expected to expel helium ashes and to retain impurities in the divertor region, if it is directed towards the divertor plate. It has been experimentally observed, however, that the flow direction is sometimes opposite; from the outer plate side to the SOL middle side in the outer SOL region of tokamaks. In order to study these SOL flow patterns by fully taking account of the kinetic effects, a full particle code, PARASOL, is applied to a tokamak plasma with the upper-null point (UN) or lower-null point (LN) divertor configuration for the downward ion ∇B drift. PARASOL simulations for the medium aspect ratio (A = 5.5) reveal the variation of the flow pattern. For the UN case with the ion ∇B drift away from the null point, the flow velocity V
∥ parallel to the magnetic field is formed almost in–out symmetrically. In the inner SOL region V
∥ is directed to the inner divertor plate and in the outer SOL V
∥ is directed to the outer plate. The stagnation point (V
∥ = 0) is located symmetrically at the bottom. On the other hand for the LN case with the ion ∇B drift towards the null point, V
∥ in the outer SOL region has a backward flow pattern. The stagnation point moves below the mid-plane of the outer SOL and V
∥ in the mid-plane outer SOL is directed to the inner plate. These simulation results are very similar to the experimental results. Simulations are carried out by changing the aspect ratio and by artificially cutting the electric field. It is found that the banana motion of trapped ions is very important for the formation of the flow pattern in addition to the self-consistent electric field. The trapped-ion effects can be stronger than the electric-field effects for the standard tokamaks with A < 5.
Dengue virus infection promotes translocation of high mobility group box 1 protein from the nucleus to the cytosol in dendritic cells, upregulates cytokine production and modulates virus replication
Authors: Kamau Edwin , Takhampunya Ratree , Li Tao , Kelly Eileen , Peachman Kristina K. , Lynch Julia A. , Sun Peifang , Palmer Dupeh R.
Published: July 2009

High mobility group box 1 (HMGB1) protein functions in regulation of transcription, cellular activation and pro-inflammatory responses. However, the potential role of HMGB1 during viral infection has not been investigated. This study attempted to elucidate whether the HMGB1-mediated inflammatory response contributes to the pathogenesis of dengue virus (DENV) infection. Our data showed that HMGB1 was released at low DENV infection levels (m.o.i. of 1) under non-necrotic conditions by human dendritic cells (DCs). When DENV-infected DCs were co-cultured with autologous T cells, there was increased production of HMGB1 by both cell types. HMGB1 regulated tumour necrosis factor alpha, interleukin (IL)-6, IL-8 and alpha interferon secretion in DENV-infected DCs. Additionally, increased HMGB1 production was associated with reduced DENV replication titres in DCs. These results suggest that HMGB1 production influences DENV infection in susceptible hosts.
Complete genomic sequences for hepatitis C virus subtypes 4b, 4c, 4d, 4g, 4k, 4l, 4m, 4n, 4o, 4p, 4q, 4r and 4t
Authors: Li Chunhua , Lu Ling , Wu Xianghong , Wang Chuanxi , Bennett Phil , Lu Teng , Murphy Donald
Published: July 2009

In this study, we characterized the full-length genomic sequences of 13 distinct hepatitis C virus (HCV) genotype 4 isolates/subtypes: QC264/4b, QC381/4c, QC382/4d, QC193/4g, QC383/4k, QC274/4l, QC249/4m, QC97/4n, QC93/4o, QC139/4p, QC262/4q, QC384/4r and QC155/4t. These were amplified, using RT-PCR, from the sera of patients now residing in Canada, 11 of which were African immigrants. The resulting genomes varied between 9421 and 9475 nt in length and each contains a single ORF of 9018–9069 nt. The sequences showed nucleotide similarities of 77.3–84.3 % in comparison with subtypes 4a (GenBank accession no. Y11604) and 4f (EF589160) and 70.6–72.8 % in comparison with genotype 1 (M62321/1a, M58335/1b, D14853/1c, and 1?/AJ851228) reference sequences. These similarities were often higher than those currently defined by HCV classification criteria for subtype (75.0–80.0 %) and genotype (67.0–70.0 %) division, respectively. Further analyses of the complete and partial E1 and partial NS5B sequences confirmed these 13 ‘provisionally assigned subtypes’.
Targeted delivery of hepatitis C virus-specific short hairpin RNA in mouse liver using Sendai virosomes
Authors: Subramanian Nithya , Mani Prashant , Roy Swagata , Gnanasundram Sivakumar Vadivel , Sarkar Debi P. , Das Saumitra
Published: July 2009

Internal ribosome entry site (IRES)-mediated translation of input viral RNA is the initial required step for the replication of the positive-stranded genome of hepatitis C virus (HCV). We have shown previously the importance of the GCAC sequence near the initiator AUG within the stem and loop IV (SLIV) region in mediating ribosome assembly on HCV RNA. Here, we demonstrate selective inhibition of HCV-IRES-mediated translation using short hairpin (sh)RNA targeting the same site within the HCV IRES. sh-SLIV showed significant inhibition of viral RNA replication in a human hepatocellular carcinoma (Huh7) cell line harbouring a HCV monocistronic replicon. More importantly, co-transfection of infectious HCV–H77s RNA and sh-SLIV in Huh7.5 cells successfully demonstrated a significant decrease in viral RNA in HCV cell culture. Additionally, we report, for the first time, the targeted delivery of sh-SLIV RNA into mice liver using Sendai virosomes and demonstrate selective inhibition of HCV-IRES-mediated translation. Results provide the proof of concept that Sendai virosomes could be used for the efficient delivery of shRNAs into liver tissue to block HCV replication.
Characterization of seven novel human papillomavirus types isolated from cutaneous tissue, but also present in mucosal lesions
Authors: de Villiers Ethel-Michele , Gunst Karin
Published: July 2009

Seven novel human papillomavirus (HPV) types were isolated and characterized. HPV 94 is related most closely to HPV 10 and belongs to the genus Alphapapillomavirus, whereas HPV 98, HPV 99, HPV 100, HPV 104, HPV 105 and HPV 113 all belong to the genus Betapapillomavirus. These HPV types were isolated from and demonstrated in cutaneous tissue, but HPV 98, HPV 100, HPV 104 and HPV 113 were also detected in malignant oesophageal and oral lesions. The general prevalence of these HPV types in lesions is infrequent.
Bahiaxenidae, a “living fossil” and a new family of Strepsiptera (Hexapoda) discovered in Brazil
Authors: Bravo Freddy , Pohl Hans , Silva‐Neto Alberto , Beutel Rolf G.
Published: July 2009

An adult male of a newly discovered strepsipteran species from Brazil—Bahiaxenos relictus—is described. A new family Bahiaxenidae is suggested based on cladistic analyses of comprehensive morphological data sets with a broad taxon sampling including the stem group. It is unambiguously placed as the sister group of all other extant families of Strepsiptera. Bahiaxenos relictus is the only species of basal, i.e. non‐stylopidian, Strepsiptera occurring in the New World. It appears to be a relict taxon that has survived in the fossil sand dunes of the São Francisco River (Bahia State). The loss of the 8th antennomere and the greatly reduced labrum are autapomorphies of Strepsiptera s.s. excluding Bahiaxenidae. The sister group relationship between†Protoxenidae and the remaining Strepsiptera, and between †Cretostylops and a clade comprising †Mengea and Strepsiptera s.s., is confirmed, as is the monophyly of Stylopidia and Stylopiformia.
Accumulation of transgene‐derived siRNAs is not sufficient for RNAi‐mediated protection against Citrus tristeza virus in transgenic Mexican lime
Authors: LÓPEZ CARMELO , CERVERA MAGDALENA , FAGOAGA CARMEN , MORENO PEDRO , NAVARRO LUIS , FLORES RICARDO , PEÑA LEANDRO
Published: July 2009

Mexican lime plants transformed with the 3′‐terminal 549 nucleotides of the Citrus tristeza virus (CTV) genome in sense, antisense and intron‐hairpin formats were analysed for transgene‐derived transcript and short interfering RNA (siRNA) accumulation, and for CTV resistance. Propagations from all sense, antisense and empty‐vector transgenic lines were susceptible to CTV, except for a single sense‐line plant with a complex transgene integration pattern that showed transgene‐derived siRNAs in association with low levels of the transgene‐derived transcript. In contrast, nine of 30 intron‐hairpin lines showed CTV resistance, with 9%–56% of bud‐propagated plants, depending on the line, remaining uninfected on graft inoculation, and the others being susceptible. Although resistance was always associated with the presence of transgene‐derived siRNAs, their level in different sense and intron‐hairpin transformants was variable irrespective of the response to CTV infection. In intron‐hairpin lines with single transgene integration, CTV resistance was correlated with low accumulation of the transgene‐derived transcript rather than with high accumulation of transgene‐derived siRNAs.
A comparison of two formulations of intradermal capsaicin as models of neuropathic pain in healthy volunteers
Authors: Gustafsson Helena , Åkesson Johanna , Lau Chai Li , Williams Desmond , Miller Lisa , Yap Sharon , Rolan Paul
Published: July 2009

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Intradermal capsaicin has some utility as a model of neuropathic pain

• Dose–response relationships have not been well documented

• Two formulations have previously been used but their performance has not been compared

WHAT THIS STUDY ADDS

• Dose–response relationships for both formulations

• The formulations are not equivalent at all doses

• The hydroxypropyl‐β‐cyclodextrin formulation is preferable

AIMS

To compare the dose–response relationships of two formulations [Tween‐ or hydroxypropyl‐β‐cyclodextrin (HP‐β‐CD)‐based] of intradermal capsaicin in healthy volunteers and to assess the effect of potential covariates of response. One, 10, 30 and 100 µg in 10 ml were compared for the outcomes of flare, spontaneous pain, mechanical allodynia and hyperalgesia in eight healthy men and eight healthy women.

RESULTS

The formulations produced comparable responses at doses 1, 10 and 30 µg, but in all parameters the response was less at 100 µg with the Tween formulation. Mean area for hyperalgesia was 9 cm2[95% confidence interval (CI) 5, 13] higher with the HP‐β‐CD formulation. Flare area was 5 cm2 (95% CI 8, 13) greater with the HP‐β‐CD formulation. There was a significant difference between pain responses from the injection site on the upper forearm compared with the lower forearm on all four pain assessments. In contrast, significant differences were seen in pain response between nondominant and dominant arm for flare, allodynia and hyperalgesia but not for spontaneous pain. A significant difference in sex was seen only for hyperalgesia. The nominal 100‐µg dose of the Tween formulation contained only 39% of label strength in the aqueous phase, which may explain the lower pharmacodynamic response.

CONCLUSION

The formulations are comparable over the dose range 1–30 µg. The significantly lower pain response at the 100 µg dose in the Tween compared with the HP‐β‐CD formulation is likely to be due to limitations in solubility at the 100 µg level. Given the greater ease of formulation and the superior dose–response relationship, the HP‐β‐CD formulation is preferable for use in the model in future studies.
Transglutaminase down‐regulates the dimerization of epidermal growth factor receptor in rat perivenous and periportal hepatocytes
Authors: Maruko A. , Ohtake Y. , Katoh S. , Ohkubo Y.
Published: July 2009

Objective:  Recently, we found that transglutaminase 2 (TG2) might be involved in the difference in proliferative capacities between periportal hepatocytes (PPH) and perivenous hepatocytes (PVH) through down‐regulation of high‐affinity epidermal growth factor receptor (EGFR). However, it is uncertain whether this high‐affinity EGFR contributes to the hepatocyte growth signalling pathway. Here, we have investigated the influence of TG2 on EGF‐induced EGFR dimerization and its phosphorylation, which are important steps in the hepatocyte proliferative/growth signalling pathway, in PPH and PVH.

Materials and methods:  PPH and PVH were isolated using the digitonin/collagenase perfusion technique. Amounts of TG2, EGFR dimerization and its phosphorylation were determined by Western blot analysis.

Results:  Pretreatment with monodansylcadaverine, an inhibitor of TG2, greatly increased EGF‐induced EGFR dimerization and its phosphorylation in PVH compared with PPH. Conversely, treatment with retinoic acid, an inducer of TG2, significantly decreased EGF‐induced EGFR dimerization and its phosphorylation with a significant increase in TG2 expression and its catalysed products, isopeptide bonds, in both subpopulations. It was found that EGFR served as a substrate for TG2.

Conclusion:  The present data showed good correlation with our previous data on EGF‐induced DNA synthesis and EGFR‐binding affinity to EGF. These results suggest that zonal difference in cell growth between PPH and PVH may be caused by down‐regulation of EGFR dimerization and subsequent autophosphorylation through TG2‐mediated cross‐linking of EGFR.
Co‐treatment with deoxycholic acid and azoxymethane accelerates secretion of HMGB1 in IEC6 intestinal epithelial cells
Authors: Fujii K. , Luo Y. , Sasahira T. , Denda A. , Ohmori H. , Kuniyasu H.
Published: July 2009

Objectives:  High‐mobility group box 1 (HMGB1) is a nuclear protein that acts as a ligand of the receptor for advanced glycation end products (RAGE) and its expression enhances progression of cancer. However, the mechanism underlying HMGB1 secretion is still unclear. In this study, we examined the effect of deoxycholic acid (DCA), a promoter of colon carcinogenesis, on HMGB1 secretion.

Materials and Methods:  We used an in vitro transformation model comprised of IEC6 intestinal epithelial cells treated with azoxymethane (AOM) and/or DCA. HMGB1 expression and secretion were examined by Western and Northern blot analyses, and ELISA. Intracellular translocation of HMGB1 was examined by protein fractionation.

Results:  AOM + DCA‐treated IEC6 cells showed upregulation of HMGB1 mRNA expression and increased level of HMGB1 protein in culture medium, but decreased level of HMGB1 protein in the nucleus. AOM + DCA treatment increased level of histone H4 acetylation, which induced translocation of HMGB1 from the nucleus to the cytoplasm and increased HMGB1 secretion. Leptomycin B inhibited extranuclear translocation and secretion of the HMGB1 protein.

Conclusion:  These findings suggest that DCA affects intracellular localization and secretion of HMGB1.
How do placements in kinship care compare with those in non‐kin foster care: placement patterns, progress and outcomes?
Authors: Farmer Elaine
Published: July 2009

As the use of kinship care is set to rise in England, it is important that policy and practice developments are based on firm evidence about kin placement outcomes and how these compare with those in stranger foster care. The research reported in this paper was based on case file reviews of 270 children, half in kin placements and half in stranger foster care, and on interviews with a sub‐sample of 32 kin carers, social workers, children and parents. Kin carers were found to be significantly more disadvantaged than stranger foster carers: more kin carers were lone carers, with health problems, living in overcrowded conditions and had financial difficulties. The children, in contrast, were remarkably similar in the two kinds of placement. The main differences between the children in the two settings are examined in the paper. The children's progress and outcomes in terms of placement quality and disruption were very similar in the two settings, but kin placements lasted longer, mainly because fewer were planned as interim placements. However, because kin carers persisted with very challenging children and yet received fewer services than stranger foster carers, they were more often under strain. The implications for policy and practice are examined.
Child well‐being through different lenses: why concept matters
Authors: Axford Nick
Published: July 2009

‘Well‐being’ has entered policy rhetoric in children's services in the UK and other Western developed countries as a companion to other buzzwords of recent years. In order to improve children's well‐being, we need not only a better understanding of what it is and how services can improve it, but also the ability to measure child well‐being in order to evaluate success. This paper explores conceptually and empirically the ‘lenses’ through which child well‐being is often viewed. It considers the relationships between these perspectives, the added value that each one brings and the implications for services.
Effects of peripheral administration of synthetic human glucose‐dependent insulinotropic peptide (GIP) on energy expenditure and subjective appetite sensations in healthy normal weight subjects and obese patients with type 2 diabetes
Authors: Daousi C. , Wilding J. P. H. , Aditya S. , Durham B. H. , Cleator J. , Pinkney J. H. , Ranganath L. R.
Published: July 2009

Background  Apart from their role in insulin secretion and glucose homeostasis, the incretin hormones glucagon‐like peptide‐1 (GLP‐1) and glucose‐dependent insulinotropic peptide (GIP) exert a number of extra‐pancreatic effects which in the case of GIP remain largely unknown.

Design/patients  Six obese male patients with diet‐controlled type 2 diabetes (T2DM) and six healthy lean male subjects were studied. The protocol included four experiments for each participant that were carried out in randomized order and included: GLP‐1 infusion at a rate of 1 pmol/kg/min for 4 h, GIP at a rate of 2 pmol/kg/min, GLP‐1 (at 1 pmol/kg/min) with GIP (at 2 pmol/kg/min), and placebo infusion for 4 h. Energy expenditure (EE) was measured throughout with indirect calorimetry and subjects were given a series of visual analogue scales to rate hourly their hunger, fullness, urge to eat and prospective consumption of food. Immediately following termination of the infusions all subjects were offered a free choice buffet lunch and total calorie and macronutrient intake was calculated.

Results  During GIP infusion there was a trend for healthy subjects to report higher hunger scores and a reduction in EE only when compared with placebo. These parameters remained unchanged in patients with T2DM. Ad libitum energy intake after all four infusions was the same in both groups.

Conclusion  We report here for the first time that GIP infusion may impact on resting EE and subjective appetite sensations in normal weight healthy subjects and further studies with larger numbers of subjects are required to help define more conclusively the precise role of GIP in energy balance in humans.
Comparison of the molecular consequences of different mutations at residue 754 and 690 of the androgen receptor (AR) and androgen insensitivity syndrome (AIS) phenotype
Authors: Tadokoro Rieko , Bunch Trevor , Schwabe John W. R. , Hughes Ieuan A. , Murphy Jane C.
Published: July 2009

Objective  Androgen insensitivity syndrome (AIS) is associated with mutations throughout the androgen receptor (AR) gene. Different mutations at the same codon have been identified in individuals with various phenotypes suggesting the nature of the codon substituted may influence the degree of AIS. We investigated if phenotype could be predicted by comparing the functionality of AR mutations with those at the same codon of known phenotype.

Patients  We identified patients from the Cambridge Disorders of Sex Development Database with the AR substitutions: Phe754Ser with microphallus without hypospadias and Asp690Val with complete AIS. Mutations Phe754Leu, Phe754Val and Asp690deletion (Asp690del) have previously been reported to be associated with different degrees of AIS.

Design  We characterized the functional properties of Phe754Ser, Phe754Leu, Phe754Val, Asp690Val and Asp690del receptor mutants in vitro and used the crystal structure of the AR ligand binding domain to model the mutations.

Results  The receptor mutants Phe754Ser, Phe754Leu and Phe754Val bound androgen with decreasing affinity, while Asp690Val showed reduced affinity compared to Asp690del. A similar pattern of reduced activation was seen on androgen responsive elements. We suggest how the mutations could affect AR structure, resulting in the observed phenotypes.

Conclusions  The relative functional properties of Phe754 and Asp690 mutant AR receptors correlate broadly with their specific phenotypes. Therefore, comparing the molecular consequences of novel mutations with others at the same codon may be a useful aid to AIS patient management, particularly for sex of rearing decisions when prediction of functionality is important.
Demographic, physical and lifestyle factors associated with androgen status: the Florey Adelaide Male Ageing Study (FAMAS)
Authors: Atlantis Evan , Martin Sean A. , Haren Matthew T. , O'Loughlin Peter D. , Taylor Anne W. , Anand‐Ivell Ravinder , Ivell Richard , Wittert Gary A. ,   Members of the Florey Adelaide Male Ageing Study
Published: July 2009

Objective  Plasma androgen levels are inversely associated with health in men, the age‐related decline of which may result from factors other than ageing per se. This study aimed to determine the effects of demographic, physical and lifestyle factors on age‐related androgen status in men.

Design  An observational survey of a regionally representative male population residing in the North West regions of Adelaide, Australia.

Participants  Study sample includes 1195 men aged 35–81 years with a response rate of 45·1%.

Measurements  Plasma levels of total testosterone (TT), bioavailable testosterone (BT), SHBG, insulin‐like peptide 3 (INSL3), and gonadotrophins were measured along with an extensive list of demographic, physical and lifestyle factors including body composition, muscle strength and biomarkers of chronic diseases, physical activity, nutrition and smoking behaviour.

Results  Low TT was mostly associated with high abdominal fat and triglycerides and low muscle strength rather than ageing per se. Low BT was associated with increased age followed by high whole body fat percentage. BT and TT levels were higher in unmarried men and smokers. SHBG levels increased with age, but were also inversely associated with insulin and triglycerides. The Leydig cell specific factor INSL3 was the strongest biomarker associated with both TT and BT.

Conclusions  Factors associated with low androgen status variably include high body fat percentage, low muscle strength and biomarkers of the metabolic syndrome. Reducing exposure to factors that adversely affect androgen status may improve the general health of ageing men by mechanisms yet to be defined.
Alkalinity generation and sediment CO2 uptake influence establishment of Sparganium angustifolium in softwater lakes
Authors: LUCASSEN ESTHER C. H. E. T. , SPIERENBURG P. , FRAAIJE R. G. A. , SMOLDERS A. J. P. , ROELOFS J. G. M.
Published: July 2009

1.  Softwater lakes are generally dominated by slow growing, small, isoetid plant species that are adapted to the carbon‐ and nutrient‐limited conditions in these lakes. We investigated the strategy of a fast growing species, Sparganium angustifolium, for occupying softwater lakes. A field survey was carried out in Norwegian carbon‐limited Isoëteto‐Lobelietum softwater lakes to compare abiotic conditions at locations with and without S. angustifolium. In addition, long term abiotic changes (1995–2008) related to the sudden establishment of the species on experimentally limed plots were studied. Based on the results, the carbon acquisition mechanism of S. angustifolium was tested in eco‐physiological laboratory experiments.

2. The redox potential was significantly lower at locations with S. angustifolium (220 ± 2.3) compared to locations without S. angustifolium (338.1 ± 13.9). The lower redox potential was accompanied by significantly higher concentrations of HCO3−, CO2 and Fe2+ in the sediment pore water, indicating in‐lake alkalinity generation due to higher iron reduction rates in the generally iron‐rich sediments. In addition, the lower redox potential was accompanied by a higher nutrient availability (NH4+ and PO43−) in the sediment pore water. Since there were no differences in water quality between the lakes, the ability of S. angustifolium to grow in softwater lakes very likely depends upon the higher dissolved inorganic carbon (DIC) and nutrient concentrations present in the sediment pore water.

3. Results from the liming experiment revealed that appearance of S. angustifolium on limed plots was related to the dissolution of Ca and Mg carbonates and development of a lower redox potential in the sediment. These processes were accompanied by a sustained increase in the availability of DIC in the sediment pore water.

4. The eco‐physiological experiments indicated that S. angustifolium can increase in biomass and produce floating leaves at a relatively high DIC availability in the root medium. In addition, it appeared that S. angustifolium can take up CO2 by the roots. As far as we know, the ability to use sediment CO2 has only been described as an adaptation typical for isoetid plant species. Use of the relatively large sediment CO2 pools present in these sediment types (>1000 μmol L−1) to enable development of long floating leaves for additional uptake of atmospheric CO2 is a very different strategy to colonise softwater lakes as compared to isoetid plant species.
Nitrogen deposition causes widespread loss of species richness in British habitats
Authors: MASKELL LINDSAY C. , SMART SIMON M. , BULLOCK JAMES M. , THOMPSON KEN , STEVENS CARLY J.
Published: July 2009

We use national scale data to test the hypothesis that nitrogen (N) deposition is strongly negatively correlated with plant species richness in a wide range of ecosystem types. Vegetation plots from a national ecological surveillance programme were drawn from heathland, acid, calcareous and mesotrophic grassland habitats. Mean species number and mean plant traits were calculated for each plot and related to atmospheric N deposition. There was a significant reduction in species richness with N deposition in acid grassland and heathland even after fitting covarying factors. In acid grassland and heathland, evidence from trait changes suggested that acidification rather than increased fertility was responsible for species loss. In contrast, calcareous grassland showed evidence of eutrophication in response to increasing N deposition. Loss of species richness from chronic N deposition is apparent in infertile grasslands and heathland. Mechanisms associated with loss of species richness differ between habitats so mitigation of N deposition should be targeted to habitat type.
The mutation spectrum associated with type 3 von Willebrand disease in a cohort of patients from the North West of England
Authors: SUTHERLAND M. S. , KEENEY S. , BOLTON‐MAGGS P. H. B. , HAY C. R. M. , WILL A. , CUMMING A. M.
Published: July 2009

Summary.  Type 3 von Willebrand disease (VWD) is a severe autosomal recessive inherited bleeding disorder. In affected individuals the underlying von Willebrand factor gene (VWF) mutations frequently remain uncharacterized. The aim of this study was to investigate the molecular basis of type 3 VWD in patients (11 Caucasians and 9 of Asian origin) attending the haemophilia centres at Central Manchester NHS Trust. A combination of DNA sequencing of VWF genomic and complementary DNA was performed to identify mutations in the patient cohort. Fifteen different VWF mutations were identified at the genomic DNA level: two gene conversion events, three nonsense, three frameshift, one missense, two splice site, one insertion‐deletion and three deletion mutations. Homozygosity or compound heterozygosity for mutations was present in 15 of the 20 patients. In the remaining five individuals, heterozygosity for a single VWF mutation was identified in four cases and one patient had no detectable VWF mutation. Analysis of platelet‐derived VWF RNA from these five individuals revealed heterozygosity for a deletion of exons 4 and 5 in four cases. The remaining patient was heterozygous for a three base deletion which had already been identified at the DNA level. Overall the observed VWF genotype explained the phenotype in 18 of the 20 patients investigated. In genetic studies in type 3 VWD, if VWF mutations are not detected at the DNA level, RNA analysis should be performed to search for intronic mutations, heterozygous deletions or aberrant splicing/post‐transcriptional events. However, this may still not explain all cases of previously phenotypically diagnosed type 3 VWD.
Low frequency of MAP kinase pathway alterations in KIT and PDGFRA wild‐type GISTs
Authors: Martinho Olga , Gouveia António , Viana‐Pereira Marta , Silva Paula , Pimenta Amadeu , Reis Rui Manuel , Lopes José Manuel
Published: July 2009

Aims:  Gastrointestinal stromal tumours (GISTs) are commonly driven by oncogenic mutations in KIT and PDGFRA. However, 10–40% of these patients are wild‐type for these genes. The prognostic significance of wild‐type GISTs is controversial, and they rarely respond to imatinib. The aim of this study was to elucidate the molecular lesions underlying wild‐type GISTs tumorigenesis.

Methods and results:  Twenty‐nine KIT and PDGFRA wild‐type GISTs were re‐assessed for the presence of ‘cryptic’KIT exon 11 duplications. Using a specific polymerase chain reaction assay, three previously undetected mutations were identified. In the remaining 26 wild‐type GISTs, KIT, stem cell factor (SCF), phospho‐KIT and phospho‐ERK expression was evaluated by immunohistochemistry. Samples were screened for gain‐of‐function mutations in the mitogen‐activated protein kinase (MAPK) cascade. KIT and SCF co‐expression associated with KIT activation was observed in approximately 30% of cases. Furthermore, phospho‐ERK expression showed that MAPK is activated in approximately 30% of cases. None of RAS family (H‐, K‐ and N‐RAS) oncogenes exhibited activating mutations, whereas BRAF mutations were found in approximately 4% of cases.

Conclusions:  In the absence of RAS mutations, MAPK could be activated through SCF/KIT autocrine/paracrine mechanisms and/or mutated BRAF in a subset of KIT/PDGFRA wild‐type GISTs.
Caveolin‐1 expression in lung carcinoma varies according to tumour histotype and is acquired de novo in brain metastases
Authors: Cassoni Paola , Daniele Lorenzo , Maldi Elena , Righi Luisella , Tavaglione Veronica , Novello Silvia , Volante Marco , Scagliotti Giorgio Vittorio , Papotti Mauro
Published: July 2009

Aims:  To study caveolin‐1 (Cav‐1) expression in metastatic lung carcinomas.

Methods and results:  Cav‐1 expression was investigated in a series of 121 lung carcinomas and it was shown that 18/121 tumours (14.9%) were Cav‐1+. None of the pure bronchioloalveolar carcinomas proved to be positive, vs. 42.8% of the large cell carcinomas (neuroendocrine subtype excluded). Adenocarcinomas (8.5%), large cell neuroendocrine carcinomas (20%) and squamous cell carcinomas (29.6%) displayed an intermediate percentage of positive cases, suggesting a gradient of Cav‐1 expression according to tumour histotype‐related aggressiveness. Moreover, the percentage of Cav‐1+ tumours with distant metastases was almost double that of non‐metastatic tumours (17.8% vs. 8.1%), irrespective of the histotype. In 34 tumours metastatic to the brain, primary and secondary lesions were compared and 53% of brain metastases were Cav‐1+ vs. 20.6% of primaries, indicating a de novo acquisition of Cav‐1 expression. This pattern was exclusive to the brain, as it was not acquired in adrenal metastases. In our series, the presence of epidermal growth factor receptor amplification, determined by fluorescence in situ hybridization, was not related to Cav‐1 reactivity.

Conclusions:  Cav‐1 immunoreactivity in lung carcinoma is histotype‐dependent and acquired de novo in brain metastases, suggesting a site‐specific phenotypic shift in secondary lesions.
Australian endodontists’ perceptions of single and multiple visit root canal treatment
Authors: Sathorn C. , Parashos P. , Messer H.
Published: July 2009

Aim  To ascertain endodontist’s point of view (treatment philosophy, rationale and preference) regarding single‐ and multiple‐visit root canal treatment. To identify the basis on which the choice is made and how the information necessary for the choice is acquired.

Methodology  Endodontists registered with the dental practice board of every state in Australia were contacted, and if they agreed to participate, they were interviewed either face to face or by telephone. The following topics were addressed in an interview lasting 15 to 20 min: demographics, current clinical procedures, treatment rationales and preference. A hypothetical scenario was posed to investigate which treatment regimen they would prefer to deliver if biological concerns were eliminated from consideration.

Results  Fifty‐two endodontists (71% of all Australian endodontists) agreed to participate in the study. Almost all (51/52) participants had performed single‐visit root canal treatment, but very few routinely performed it. A majority of participants were willing to provide single‐visit treatment where patients had time constraints, and in vital cases (including elective endodontics). The most powerful factor of influencing practice change was interpersonal contact with colleagues. Publications in academic journals have a weak influence in practice change.

Conclusions  Australian endodontists strongly prefer multiple‐visit over single‐visit root canal treatment even in cases where biological concerns are not an issue. Operator preference rather than biological or patient considerations appear to be the primary determinant of treatment choice.