bluuebunny/crossref_metadata_2025 huggingface dataset:
Why 0.02%? A review of the basis for current practice in aerial 1080 baiting for rabbits in New Zealand
Authors: Nugent Graham , Twigg Laurie E. , Warburton Bruce , McGlinchy Aaron , Fisher Penny , Gormley Andrew M. , Parkes John P.
Published: April 2012

Sodium fluoroacetate (1080) has been used as an aerially distributed toxin against mammalian pests in New Zealand since the 1950s. Although its use for rabbit (Oryctolagus cuniculus) control ceased temporarily after the illegal release of rabbit haemorrhagic disease virus (RHDV) in 1997, there has been a recent resurgence in the use of aerial baiting with 1080 to control rabbits as the efficacy of RHDV has fallen. Current practices for rabbit control using 1080 have changed little since the 1980s, with high sowing rates and low toxin loadings commonplace. The lack of ongoing development in baiting practices for rabbit control contrasts sharply with continued improvements in the aerial 1080 baiting practices for brushtail possums (Trichosurus vulpecula) in New Zealand, such as a shift to a comparatively high 1080 loading and using much lower application rates of prefeed and toxic bait. These modifications have resulted in an overall reduction in the amount of toxin used for possum control. The disparity in these two approaches prompted a formal review of the rationale on which the current 1080 baiting practices for rabbits are based. Two issues emerged strongly. First, the current low toxin loading used (0.02–0.04% 1080 in bait) is not based on experimental optimisation in New Zealand but, rather, on research conducted several decades earlier in Australia. Second, despite long-standing concerns about the quality of carrot bait used in New Zealand, current bait manufacturing and distribution practices still produce large numbers of small sublethal fragments. Thus, the current New Zealand practice of multiple prefeeds and very high sowing rates of bait with a low 1080 loading used against rabbits seems to have resulted from the need to compensate for the low toxic loading and poor quality control of the bait (carrots). We, therefore, suggest that there is considerable potential to improve current aerial 1080 baiting practices for controlling rabbits in New Zealand. More generally, these findings also help illustrate that ‘best’ pest-management practice may sometimes be based on pragmatic solutions aimed at overcoming unrecognised internal constraints that are in fact avoidable. Refining and modernising vertebrate pest-control programs, so that they better meet efficacy requirements and contemporary public expectations, therefore requires understanding not just that a solution works, but also how it works.
Microfluidic integration for automated targeted proteomic assays
Authors: Hughes Alex J. , Lin Robert K. C. , Peehl Donna M. , Herr Amy E.
Published: April 2012

A dearth of protein isoform-based clinical diagnostics currently hinders advances in personalized medicine. A well-organized protein biomarker validation process that includes facile measurement of protein isoforms would accelerate development of effective protein-based diagnostics. Toward scalable protein isoform analysis, we introduce a microfluidic “single-channel, multistage” immunoblotting strategy. The multistep assay performs all immunoblotting steps: separation, immobilization of resolved proteins, antibody probing of immobilized proteins, and all interim wash steps. Programmable, low-dispersion electrophoretic transport obviates the need for pumps and valves. A three-dimensional bulk photoreactive hydrogel eliminates manual blotting. In addition to simplified operation and interfacing, directed electrophoretic transport through our 3D nanoporous reactive hydrogel yields superior performance over the state-of-the-art in enhanced capture efficiency (on par with membrane electroblotting) and sparing consumption of reagents (
ca.
1 ng antibody), as supported by empirical and by scaling analyses. We apply our fully integrated microfluidic assay to protein measurements of endogenous prostate specific antigen isoforms in (
i
) minimally processed human prostate cancer cell lysate (1.1 pg limit of detection) and (
ii
) crude sera from metastatic prostate cancer patients. The single-instrument functionality establishes a scalable microfluidic framework for high-throughput targeted proteomics, as is relevant to personalized medicine through robust protein biomarker verification, systematic characterization of new antibody probes for functional proteomics, and, more broadly, to characterization of human biospecimen repositories.
In vivo imaging of lymphatic vessels in development, wound healing, inflammation, and tumor metastasis
Authors: Martínez-Corral Inés , Olmeda David , Diéguez-Hurtado Rodrigo , Tammela Tuomas , Alitalo Kari , Ortega Sagrario
Published: April 2012

Lymphatic vessel growth or lymphangiogenesis occurs during embryonic development and wound healing and plays an important role in tumor metastasis and inflammatory diseases. However, the possibility of noninvasive detection and quantification of lymphangiogenesis has been lacking. Here, we present theVegfr3EGFPLucmouse model, where an EGFP-luciferase fusion protein, expressed under the endogenous transcriptional control of theVegfr3gene, allows the monitoring of physiological and pathological lymphangiogenesis in vivo. We show tracking of lymphatic vessel development during embryogenesis as well as lymphangiogenesis induced by specific growth factors, during wound healing and in contact hypersensitivity (CHS) - induced inflammation where we also monitor down-regulation of lymphangiogenesis by the glucocorticoid dexamethasone. Importantly, theVegfr3-reporter allowed us to tracking tumor-induced lymphangiogenesis at the tumor periphery and in lymph nodes in association with the metastatic process. This is the first reporter mouse model for luminescence imaging of lymphangiogenesis. It should provide an important tool for studying the involvement of lymphangiogenesis in pathological processes.
Induction of lignocellulose-degrading enzymes in
Neurospora crassa
by cellodextrins
Authors: Znameroski Elizabeth A. , Coradetti Samuel T. , Roche Christine M. , Tsai Jordan C. , Iavarone Anthony T. , Cate Jamie H. D. , Glass N. Louise
Published: April 2012

Neurospora crassa
colonizes burnt grasslands in the wild and metabolizes both cellulose and hemicellulose from plant cell walls. When switched from a favored carbon source such as sucrose to cellulose,
N. crassa
dramatically upregulates expression and secretion of a wide variety of genes encoding lignocellulolytic enzymes. However, the means by which
N. crassa
and other filamentous fungi sense the presence of cellulose in the environment remains unclear. Here, we show that an
N. crassa
mutant carrying deletions of two genes encoding extracellular β-glucosidase enzymes and one intracellular β-glucosidase lacks β-glucosidase activity, but efficiently induces cellulase gene expression in the presence of cellobiose, cellotriose, or cellotetraose as a sole carbon source. These data indicate that cellobiose, or a modified version of cellobiose, functions as an inducer of lignocellulolytic gene expression in
N. crassa
. Furthermore, the inclusion of a deletion of the catabolite repressor gene,
cre-1
, in the triple β-glucosidase mutant resulted in a strain that produces higher concentrations of secreted active cellulases on cellobiose. Thus, the ability to induce cellulase gene expression using a common and soluble carbon source simplifies enzyme production and characterization, which could be applied to other cellulolytic filamentous fungi.
Small molecule inhibitors of influenza A and B viruses that act by disrupting subunit interactions of the viral polymerase
Authors: Muratore Giulia , Goracci Laura , Mercorelli Beatrice , Foeglein Ágnes , Digard Paul , Cruciani Gabriele , Palù Giorgio , Loregian Arianna
Published: April 2012

Influenza viruses are the cause of yearly epidemics and occasional pandemics that represent a significant challenge to public health. Current control strategies are imperfect and there is an unmet need for new antiviral therapies. Here, we report the identification of small molecule compounds able to effectively and specifically inhibit growth of influenza A and B viruses in cultured cells through targeting an assembly interface of the viral RNA-dependent RNA polymerase. Using an existing crystal structure of the primary protein–protein interface between the PB1 and PA subunits of the influenza A virus polymerase, we conducted an in silico screen to identify potential small molecule inhibitors. Selected compounds were then screened for their ability to inhibit the interaction between PB1 and PA in vitro using an ELISA-based assay and in cells, to inhibit nuclear import of a binary PB1–PA complex as well as transcription by the full viral ribonucleoprotein complex. Two compounds emerged as effective inhibitors with IC
50
values in the low micromolar range and negligible cytotoxicity. Of these, one compound also acted as a potent replication inhibitor of a variety of influenza A virus strains in Madin-Darby canine kidney (MDCK) cells, including H3N2 and H1N1 seasonal and 2009 pandemic strains. Importantly, this included an oseltamivir-resistant isolate. Furthermore, potent inhibition of influenza B viruses but not other RNA or DNA viruses was seen. Overall, these compounds provide a foundation for the development of a new generation of therapeutic agents exhibiting high specificity to influenza A and B viruses.
Fukushima-derived radionuclides in the ocean and biota off Japan
Authors: Buesseler Ken O. , Jayne Steven R. , Fisher Nicholas S. , Rypina Irina I. , Baumann Hannes , Baumann Zofia , Breier Crystaline F. , Douglass Elizabeth M. , George Jennifer , Macdonald Alison M. , Miyamoto Hiroomi , Nishikawa Jun , Pike Steven M. , Yoshida Sashiko
Published: April 2012

The Tōhoku earthquake and tsunami of March 11, 2011, resulted in unprecedented radioactivity releases from the Fukushima Dai-ichi nuclear power plants to the Northwest Pacific Ocean. Results are presented here from an international study of radionuclide contaminants in surface and subsurface waters, as well as in zooplankton and fish, off Japan in June 2011. A major finding is detection of Fukushima-derived
134
Cs and
137
Cs throughout waters 30–600 km offshore, with the highest activities associated with near-shore eddies and the Kuroshio Current acting as a southern boundary for transport. Fukushima-derived Cs isotopes were also detected in zooplankton and mesopelagic fish, and unique to this study we also find
110m
Ag in zooplankton. Vertical profiles are used to calculate a total inventory of ∼2 PBq
137
Cs in an ocean area of 150,000 km
2
. Our results can only be understood in the context of our drifter data and an oceanographic model that shows rapid advection of contaminants further out in the Pacific. Importantly, our data are consistent with higher estimates of the magnitude of Fukushima fallout and direct releases [Stohl et al. (2011)
Atmos Chem Phys Discuss
11:28319–28394; Bailly du Bois et al. (2011)
J Environ Radioact
, 10.1016/j.jenvrad.2011.11.015]. We address risks to public health and marine biota by showing that though Cs isotopes are elevated 10–1,000× over prior levels in waters off Japan, radiation risks due to these radionuclides are below those generally considered harmful to marine animals and human consumers, and even below those from naturally occurring radionuclides.
Biotinylation of lysine method identifies acetylated histone H3 lysine 79 in
Saccharomyces cerevisiae
as a substrate for Sir2
Authors: Bheda Poonam , Swatkoski Stephen , Fiedler Katherine L. , Boeke Jef D. , Cotter Robert J. , Wolberger Cynthia
Published: April 2012

Although the biological roles of many members of the sirtuin family of lysine deacetylases have been well characterized, a broader understanding of their role in biology is limited by the challenges in identifying new substrates. We present here an in vitro method that combines biotinylation and mass spectrometry (MS) to identify substrates deacetylated by sirtuins. The method permits labeling of deacetylated residues with amine-reactive biotin on the ϵ-nitrogen of lysine. The biotin can be utilized to purify the substrate and identify the deacetylated lysine by MS. The biotinyl-lysine method was used to compare deacetylation of chemically acetylated histones by the yeast sirtuins, Sir2 and Hst2. Intriguingly, Sir2 preferentially deacetylates histone H3 lysine 79 as compared to Hst2. Although acetylation of K79 was not previously reported in
Saccharomyces cerevisiae
, we demonstrate that a minor population of this residue is indeed acetylated in vivo and show that Sir2, and not Hst2, regulates the acetylation state of H3 lysine 79. The in vitro biotinyl-lysine method combined with chemical acetylation made it possible to identify this previously unknown, low-abundance histone acetyl modification in vivo. This method has further potential to identify novel sirtuin deacetylation substrates in whole cell extracts, enabling large-scale screens for new deacetylase substrates.
Biosynthetic pathway toward carbohydrate-like moieties of alnumycins contains unusual steps for C-C bond formation and cleavage
Authors: Oja Terhi , Klika Karel D. , Appassamy Laura , Sinkkonen Jari , Mäntsälä Pekka , Niemi Jarmo , Metsä-Ketelä Mikko
Published: April 2012

Carbohydrate moieties are important components of natural products, which are often imperative for the solubility and biological activity of the compounds. The aromatic polyketide alnumycin A contains an extraordinary sugar-like 4′-hydroxy-5′-hydroxymethyl-2′,7′-dioxane moiety attached via a carbon-carbon bond to the aglycone. Here we have extensively investigated the biosynthesis of the dioxane unit through
13
C labeling studies, gene inactivation experiments and enzymatic synthesis. We show that AlnA and AlnB, members of the pseudouridine glycosidase and haloacid dehalogenase enzyme families, respectively, catalyze C-ribosylation conceivably through Michael-type addition of
d
-ribose-5-phosphate and dephosphorylation. The ribose moiety may be attached both in furanose (alnumycin C) and pyranose (alnumycin D) forms. The C

1
′

-C

2
′

bond of alnumycin C is subsequently cleaved and the ribose unit is rearranged into an unprecedented dioxolane (
cis
-bicyclo[3.3.0]-2′,4′,6′-trioxaoctan-3′β-ol) structure present in alnumycin B. The reaction is catalyzed by Aln6, which belongs to a previously uncharacterized enzyme family. The conversion was accompanied with consumption of O
2
and formation of H
2
O
2
, which allowed us to propose that the reaction may proceed via hydroxylation of C1′ followed by retro-aldol cleavage and acetal formation. Interestingly, no cofactors could be detected and the reaction was also conducted in the presence of metal chelating agents. The last step is the conversion of alnumycin B into the final end-product alnumycin A catalyzed by Aln4, an NADPH-dependent aldo-keto reductase. This characterization of the dioxane biosynthetic pathway sets the basis for the utilization of C-C bound ribose, dioxolane and dioxane moieties in the generation of improved biologically active compounds.
Murine gammaherpesvirus 68 infection protects lupus-prone mice from the development of autoimmunity
Authors: Larson Jennifer D. , Thurman Joshua M. , Rubtsov Anatoly V. , Claypool David , Marrack Philippa , van Dyk Linda F. , Torres Raul M. , Pelanda Roberta
Published: April 2012

Gammaherpesvirus infections, such as those caused by EBV, have been suggested to promote the development of autoimmunity. To test this idea, we infected healthy WT and lupus-prone B6.Sle123 mice with an EBV-related and rodent-specific gammaherpesvirus, γHV68. Although acute γHV68 infection increased autoantibody levels for 4 to 6 wk, latent infection inhibited these responses for 1 y. The inhibition of autoantibody expression was only observed in B6.Sle123 females and not in males, which already displayed lower autoantibody titers. Contrary to the initial hypothesis, infection of young B6.Sle123 mice, both male and female, resulted in suppression of lymphoid activation and expansion and of glomerular inflammation and sclerosis, preserving kidney function. Moreover, γHV68 infection led to reduced autoantibody titers, lymphoid activation, and glomerular inflammation whether lupus-prone females were infected before or during disease manifestation. Finally, γHV68 infection also inhibited autoantibody production in the genetically distinct MRL/
lpr
lupus-prone mice. Our findings indicate that γHV68 infection strongly inhibits the development and progression of lupus-like disease in mice that spontaneously develop this condition mediating its beneficial effects at the humoral, cellular, and organ levels. The mechanisms by which the virus exerts this down-modulatory action are not yet clear, but appear to operate via reduced activation of dendritic cells, T cells, and B cells. Gammaherpesviruses coevolved with the vertebrate immune systems, establishing lifelong infections in humans and other mammals. Our findings that γHV68 infection prevents rather than exacerbates autoimmunity in mice suggest that infection with gammaherpesviruses may be protective rather than pathological in most individuals.
Leptin Is an Endogenous Protective Protein against the Toxicity Exerted by Tumor Necrosis Factor
Authors: Takahashi Nozomi , Waelput Wim , Guisez Yves
Published: April 2012

Tumor necrosis factor (TNF) is a central mediator of a number of important pathologies such as the systemic inflammatory response syndrome. Administration of high TNF doses induces acute anorexia, metabolic derangement, inflammation, and eventually shock and death. The in vivo effects of TNF are largely mediated by a complex network of TNF-induced cytokines and hormones acting together or antagonistically. Since TNF also induces leptin, a hormone secreted by adipocytes that modulates food intake and metabolism, we questioned the role of leptin in TNF-induced pathology. To address this question, we tested mouse strains that were defective either in leptin gene (ob/ob) or in functional leptin receptor gene (db/db), and made use of a receptor antagonist of leptin. Ob/ob and db/db mice, as well as normal mice treated with antagonist, exhibited increased sensitivity to the lethal effect of TNF. Exogenous leptin afforded protection to TNF in ob/ob mice, but failed to enhance the protective effect of endogenous leptin in normal mice. We conclude that leptin is involved in the protective mechanisms that allow an organism to cope with the potentially autoaggressive effects of its immune system.
Postcolonial Perspectives on Early Modern Canada: Champlain's
Voyages de la Nouvelle France
(1632)
Authors: Haskins Gonthier Ursula
Published: April 2012

The early modern dimension of francophone postcolonialism has yet to be fully explored. This paper adds historical depth to postcolonial studies by applying postcolonial theory to an early seventeenth-century French colonial text: Samuel de Champlain's
Voyages de la Nouvelle France
(1632). A postcolonial reading of Champlain's
Voyages
reveals the ambivalence latent in a text traditionally seen as a straightforward narrative of the coming of French civilization to the Americas. After evaluating the applicability of postcolonial theory to works such as Champlain's
Voyages
, discussion focuses on an episode from the text in which, on the eve of a battle against the Iroquois, Champlain has a dream predicting the defeat of the enemy by his Huron allies. As the author had previously dismissed, with sceptical superiority, the Amerindian faith in portentous dreams, this apparent shift in cultural allegiance raises interesting questions. Postcolonialism's insights into colonial power relations can shed new light on Champlain's complex relationship with his Amerindian allies and on the identity issues faced by colonizers in ‘white settler’ colonies. This paper traces Champlain's transformation from Frenchman in Canada to incipient French-Canadian, and ultimately tests the limits of taking a postcolonial perspective on the early modern.
A Novel Lactone-Forming Carboxylesterase: Molecular Identification of a Tuliposide A-Converting Enzyme in Tulip
Authors: Nomura Taiji , Ogita Shinjiro , Kato Yasuo
Published: April 2012

Tuliposides, the glucose esters of 4-hydroxy-2-methylenebutanoate and 3,4-dihydroxy-2-methylenebutanoate, are major secondary metabolites in tulip (Tulipa gesneriana). Their lactonized aglycons, tulipalins, function as defensive chemicals due to their biological activities. We recently found that tuliposide-converting enzyme (TCE) purified from tulip bulbs catalyzed the conversion of tuliposides to tulipalins, but the possibility of the presence of several TCE isozymes was raised: TCE in tissues other than bulbs is different from bulb TCE. Here, to prove this hypothesis, TCE was purified from petals, which have the second highest TCE activity after bulbs. The purified enzyme, like the bulb enzyme, preferentially accepted tuliposides as substrates, with 6-tuliposide A the best substrate, which allowed naming the enzyme tuliposide A-converting enzyme (TCEA), but specific activity and molecular mass differed between the petal and bulb enzymes. After peptide sequencing, a novel cDNA (TgTCEA) encoding petal TCEA was isolated, and the functional characterization of the recombinant enzyme verified that TgTCEA catalyzes the conversion of 6-tuliposide A to tulipalin A. TgTCEA was transcribed in all tulip tissues but not in bulbs, indicating the presence of a bulb-specific TgTCEA, as suggested by the distinct enzymatic characters between the petal and bulb enzymes. Plastidial localization of TgTCEA enzyme was revealed, which allowed proposing a cytological mechanism of TgTCE-mediated tulipalin formation in the tulip defensive strategy. Site-directed mutagenesis of TgTCEA suggested that the oxyanion hole and catalytic triad characteristic of typical carboxylesterases are essential for the catalytic process of TgTCEA enzyme. To our knowledge, TgTCEA is the first identified member of the lactone-forming carboxylesterases, specifically catalyzing intramolecular transesterification.
Potential antitumor applications of a monoclonal antibody specifically targeting human papilloma virus 16 E749–57 peptide
Authors: Lei Jianqiang , Zhang GaoXia
Published: April 2012

Our study aims to evaluate whether the approach of TCRm mAb has therapeutic potential against HPV‐induced tumors. In the present study, we generated a murine IgG2a mAb 6C10 specifically recognizing HPV‐16‐E749–57 epitope (RAHYNIVTF) in the polypeptides and in complex with a MHC class I molecule. Analysis of the primary structure shows that the 6C10 Ab displays a novel sequence in the CDR of the heavy chain, compared to the sequences in the Kabat database, which suggests the Ab has completed its affinity maturation. The 6C10 Ab can specifically recognize E7 and Trx‐E730–67 protein in ELISA, and can also specifically bind to T2 cell carrying HPV‐16‐E749–57 peptide. In the TC‐1 cell tumor‐bearing mouse model, 6C10 exhibits tumor suppression activity when compared to the isotype control Ab. 6C10 Ab has showed tumor‐inhibition potency in a mouse model and this Ab may have the prospect of cancer therapy.
Notch1 activation promotes renal cell carcinoma growth via PI3K/Akt signaling
Authors: Xu Le , Zhu Yu , Xu Jiejie , Wu Kerong , Li Jinfeng , Xu Wenping , Liu Haiou , Wang Shanshan , Yin Hankun , Chen Lin , Wang Guomin , Lin Zongming
Published: April 2012

Both the Notch1 and PI3K/Akt pathways are aberrantly activated in clear cell renal cell carcinoma (CCRCC) and involved in the tumorigenesis. The aim of this study was to test our hypothesis that elevated Notch1 signaling activity exerts its growth‐promoting effects via the PI3K/Akt pathway in CCRCC. To investigate the relationship between the two pathways, we enhanced and suppressed the Notch1 activity respectively in a CCRCC cell line through diverse means, and then evaluated ensuing phosphorylated Akt (pAkt) levels. To further study their collaboration in promoting tumor growth, cell proliferation assay, colony formation assay and cell cycle analysis were conducted under several different conditions. Immunostaining of the tissue microarrays was used to determine whether the phenomena we observed also existed in vivo. The results showed that Notch1 signaling was activated in CCRCC tissue samples and cell lines. Notch1 activation increased CCRCC cell proliferation, enhanced anchorage‐independent growth, and accelerated G1/S cell cycle progression. Such effects of the Notch1 signaling were, at least in part, mediated by the PI3K/Akt pathway. Correlations between Notch1, pAkt and Ki‐67 protein levels in tissue microarrays reinforced our in vitro findings. Taken together, the current study established a functional link between the Notch1 and PI3K/Akt pathways in CCRCC. (Cancer Sci 2012; 103: 1253–1258)
Is Helicobacter pylori antibiotic resistance surveillance needed and how can it be delivered?
Authors: McNulty C. A. M. , Lasseter G. , Shaw I. , Nichols T. , D'Arcy S. , Lawson A. J. , Glocker E.
Published: April 2012

Background

Most patients are prescribed Helicobacter pylori treatment without culture and antibiotic susceptibility testing, as current guidance recommends that patients with recurrent dyspepsia should be tested for H. pylori using a non‐invasive breath or faecal antigen test.

Aims

To determine the prevalence of H. pylori antibiotic resistance in patients attending endoscopy in England and Wales, and the feasibility of an antibiotic resistance surveillance programme testing.

Methods

We tested the antibiotic susceptibility of H. pylori isolates from biopsy specimens from 2063 of 7791 (26%) patients attending for endoscopy in Gloucester and Bangor, and 339 biopsy specimens sent to the Helicobacter Reference Unit (HRU) in London. Culture and susceptibility testing was undertaken in line with National and European methods.

Results

Helicobacter pylori were cultured in 6.4% of 2063 patients attending Gloucester and Bangor hospitals. Resistance to amoxicillin, tetracycline and rifampicin/rifabutin was below 3% at all centres. Clarithromycin, metronidazole and quinolone resistance was significantly higher in HRU (68%, 88%, 17%) and Bangor isolates (18%, 43%, 13%) than Gloucester (3%, 22%, 1%). Each previous course of these antibiotics is associated with an increase in the risk of antibiotic resistance to that agent [clarithromycin: RR = 1.5 (P = 0.12); metronidazole RR = 1.6 (P = 0.002); quinolone RR = 1.8 (P = 0.01)].

Conclusions

Helicobacter pylori infection is now uncommon in dyspeptic patients at endoscopy. A surveillance system is feasible and necessary to inform dyspepsia management guidance. Clinicians should take a thorough antibiotic history before prescribing metronidazole, clarithromycin or levofloxacin for H. pylori.
Genetic lesions of the TRAF3 and MAP3K14 genes in classical Hodgkin lymphoma
Authors: Otto Claudia , Giefing Maciej , Massow Anne , Vater Inga , Gesk Stefan , Schlesner Matthias , Richter Julia , Klapper Wolfram , Hansmann Martin‐Leo , Siebert Reiner , Küppers Ralf
Published: April 2012

Hodgkin and Reed/Sternberg (HRS) cells in classical Hodgkin lymphoma (cHL) show constitutive activation of nuclear factor (NF)‐κB. Several genetic lesions contribute to this deregulated NF‐κB activity. Here, we analysed two further NF‐κB regulators for genetic lesions, the inhibitory factor TRAF3 and the key signalling component of the alternative NF‐κB pathway, MAP3K14 (NIK). Single nucleotide polymorphism (SNP) array analysis of cHL cell lines revealed a uniparental disomy of the long arm of chromosome 14 associated with a biallelic deletion of TRAF3 located on this chromosome in cell line U‐HO1. Cloning of the deletion breakpoint showed a 123 371 bp deletion. No inactivating mutations of TRAF3 were found in six other cHL cell lines or in microdissected HRS cells from seven cHL. However, in primary cHL samples interphase cytogenetic analyses revealed signal patterns indicating monoallelic deletion of TRAF3 in 3/20 other cases. SNP array analysis revealed a gain of copy number for MAP3K14 in three cHL cell lines. Gains of MAP3K14 were detected in 5/16 cases of primary cHL. In conclusion, in rare instances, HRS cells harbour inactivating mutations of the TRAF3 gene and recurrently show gains of MAP3K14, indicating that more components of NF‐κB signalling show genetic lesions in HRS cells than previously known.
Sox2 targets cyclinE, p27 and survivin to regulate androgen‐independent human prostate cancer cell proliferation and apoptosis
Authors: Lin F. , Lin P. , Zhao D. , Chen Y. , Xiao L. , Qin W. , Li D. , Chen H. , Zhao B. , Zou H. , Zheng X. , Yu X.
Published: April 2012

Objectives

Sox2 is a major transcription factor and the transforming growth factor‐α (TGF‐α)/EGFR autocrine loop is a hallmark of prostate cancer progression. In this study, we have evaluated the effects and potential mechanisms of Sox2 on cell proliferation and apoptosis, and investigated effects of TGF‐α on expression of Sox2 on androgen‐independent human prostate cancer cells.

Materials and methods

Expression of Sox2 has been determined by RT‐PCR, western blot analysis and immunocytochemistry, using RNAi and over‐expression strategy to study functions of Sox2 in DU145 and PC‐3 cells. Changes in level of proliferation, cell cycle and apoptosis profiles were measured by MTT, colony‐forming, bromodeoxyuridine incorporation assays, cell cycle and annexin V analysis.

Results

Sox2 was expressed in six human prostate cancer cell lines, and its inhibition reduced cell proliferation and induced apoptosis in DU145 cells. We have shown that knock‐down of Sox2 inhibited G1 to S phase transition concomitantly with down‐regulation of cyclin E and up‐regulation of p27 proteins. Conversely, over‐expression of Sox2 led to the opposite effect in PC‐3 cells but its inhibition induced apoptosis by down‐regulation of survivin in DU145 cells. We also found that TGF‐α up‐regulated Sox2 and survivin protein expression via the EGFR/PI3K/AKT pathway.

Conclusions

Sox2 expression is necessary for cell proliferation and evasion of apoptosis in prostate cancer cells and TGF‐α could regulate Sox2 and survivin expression by activating the EGFR/PI3K/AKT pathway.
Inhibition of phosphodiestrase 9 induces cGMP accumulation and apoptosis in human breast cancer cell lines, MCF‐7 and MDA‐MB‐468
Authors: Saravani R. , Karami‐Tehrani F. , Hashemi M. , Aghaei M. , Edalat R.
Published: April 2012

Objectives

Phosphodiesterase 9 (PDE9) is a major isoform of phosphodiesterase hydrolysing cGMP and plays a key role in proliferation of cells, their differentiation and apoptosis, via intracellular cGMP signalling. The study described here was designed to investigate expression, activity and apoptotic effect of PDE9 on human breast cancer cell lines, MCF‐7 and MDA‐MB‐468.

Materials and methods

Activity and expression of PDE9 were examined using colorimetric cyclic nucleotide phosphodiesterase assay and real‐time RT‐PCR methods respectively; cGMP concentration was also measured. MTT viability test, annexin V‐FITC staining, Hoechst 33258 staining and caspase3 activity assay were used to detect apoptosis.

Results

Treatment of both cell lines with BAY 73‐6691 lead to reduction in PDE9 mRNA expression, PDE9 cGMP‐hydrolytic activity and elevation of the intracellular cGMP response. BAY 73‐6691 significantly reduced cell proliferation in a dose‐ and time‐dependent manner and caused marked increase in apoptosis through caspase3 activation.

Conclusion

Our results revealed that BAY 73‐6691 induced apoptosis in these breast cancer cell lines through the cGMP pathway. These data suggest that BAY 73‐6691 could be utilized as an agent in treatment of breast cancer.
A relationship‐based approach to engaging involuntary clients: the contribution of recognition theory
Authors: Turney Danielle
Published: April 2012

This paper focuses on the process of engaging with families where a child is at risk of harm, and considers a relationship‐based approach to work with ‘involuntary clients’ of child protection services. Contextualizing the discussion within a broader understanding of the role and importance of relationship‐based practice, a conceptual and ethical framework is outlined that can, it is suggested, support effective relationship‐based work and moral decision‐making with involuntary clients. Making use of psychoanalytic and more broadly psychodynamic insights, along with perspectives drawn from recognition theory, it is argued that relationship‐based practice offers the potential for recognition, respect and reciprocity, and that these three aspects of relationship provide a foundation for ethical engagement with involuntary clients. However, such an approach is not without tensions, so the latter part of the paper considers some of the challenges and dilemmas that accompany the process of trying to engage parents who do not want to be ‘worked with’.
‘I'm just a mother. I'm nothing special, they're all professionals’: parental advocacy as an aid to parental engagement
Authors: Featherstone Brid , Fraser Claire
Published: April 2012

This paper discusses findings from an evaluation of an advocacy scheme for parents whose children were subject to child protection proceedings including a pilot project where co‐operation between parents and professionals was an issue. Encouraging findings of parents' experiences of being heard, being able to listen, and, in some cases, to act on concerns about their children's welfare are outlined. Three case studies offer accounts of practices that are gendered and occur against a backdrop of ethnic disadvantage. The paper suggests that current debates about reform need to recognize the need to support parents to manage intimidating systems.
Evaluation of residential care from the perspective of older adolescents in care. The need for a new construct: optimum professional proximity
Authors: Soldevila Anna , Peregrino Antonio , Oriol Xavier , Filella Gemma
Published: April 2012

This paper analyses the evaluation of residential care from the perspective of the adolescents in care. Twelve discussion groups were carried out in 12 different centres in Catalonia (Spain). A total of 66 adolescents aged between 16 and 18 years of age participated. The data contributed by the discussion groups were analysed and categorized using descriptive analysis matrices. Most notable from the results is that the majority of those interviewed call for increased emotional involvement from the social workers, more contact with their families and re‐educational intervention within their families, which renders the return to these possible within a shorter period of time. Data provided by the adolescents lead us to reconsider the concept of optimum professional distance and to suggest replacing it with optimum professional proximity; this would be based on emotional involvement and not on emotional distance, although not on emotional dependence either.
Bronchial inflammation and hyperresponsiveness in well controlled asthma
Authors: Muñoz X. , Sanchez‐Vidaurre S. , Roca O. , Torres F. , Morell F. , Cruz M. J.
Published: April 2012

Background

Little research has been devoted to the characteristics of bronchial inflammation in patients with stable, well controlled asthma.

Objective

The aim of this study was to assess the degree and type of airway inflammation and to investigate the relationship between inflammation and bronchial hyperresponsiveness in patients with well controlled asthma.

Methods

A cross‐sectional study was conducted in 84 adult patients (43 men, mean age 43 years) with documented well controlled asthma. Induced sputum samples were obtained and cell types determined by differential cell count. Spirometry and methacholine challenge testing were performed. Asthma Control Questionnaire (ACQ) was used to assess symptoms. Patients were included if their ACQ score was < 0.75.

Results

A total of 59 patients had persistent bronchial inflammation: 28 cases were considered eosinophilic, 28 neutrophilic, and 3 mixed. Median (range) percentage of eosinophils was 4% (0–64) in patients testing positive to methacholine challenge (n = 66) and 1% (0–3) in those testing negative (n = 18) (P = 0.003). A positive correlation was found between eosinophil percentage and the methacholine dose/response ratio (r = 0.477, P = 0.0001). The geometric mean (95% CI) of the methacholine PC20 was 1.74 mg/mL (1.04–2.93) in patients with eosinophilic inflammation and 4.14 mg/mL (2.5–6.84) in those with neutrophilic inflammation (P = 0.03).

Conclusions

Inflammation and bronchial hyperresponsiveness persist in most patients with well controlled asthma.

Clinical Relevance

The study demonstrates that eosinophilic or neutrophilic inflammation persisted in most well controlled asthma patients despite the fact that their condition was controlled and therefore, measurement of bronchial inflammation seems essential to achieve proper asthma control.
Childhood wheezing phenotypes and FeNO in atopic children at age 8
Authors: van der Valk R. J. P. , Caudri D. , Savenije O. , Koppelman G. H. , Smit H. A. , Wijga A. H. , Postma D. S. , Kerkhof M. , Brunekreef B. , de Jongste J. C.
Published: April 2012

Background

Fractional exhaled Nitric Oxide (FeNO) is a surrogate biomarker of the degree of eosinophilic airway inflammation. Using longitudinal latent class analysis, five wheezing phenotypes have been identified, characterized by different ages of onset and prognosis.

Objectives

To assess FeNO measured at 4 and 8 years in children with different phenotypes of wheeze and atopy.

Methods

Children participated in the Prevention and Incidence of Asthma and Mite Allergy (PIAMA) study, a prospective birth cohort in the Netherlands. Respiratory health was assessed yearly by questionnaires until the age of 8 years; these data were used to identify five wheezing phenotypes. Associations between FeNO and wheezing phenotypes were investigated using weighted linear regression.

Results

Data on wheezing phenotypes and FeNO at 4 and 8 years were available in 588 and 973 children respectively. Compared with the phenotype of never and transient wheeze, FeNO at 4 years was higher in intermediate onset and persistent wheeze. FeNO at 8 years of age differed significantly between all phenotypes, with highest FeNO values for persistent, intermediate onset, and late onset wheeze. Rise in FeNO from 4 to 8 years in intermediate and late onset wheezers was significantly higher compared to FeNO rise in never and transient wheezers. Stratified analyses showed that the increase in FeNO in persistent, intermediate, and late onset wheeze was only present in children with allergic sensitization at 8 years.

Conclusions and Clinical Relevance

The FeNO measured at 8 years was associated with specific wheezing phenotypes, only among atopic children.
Thyrotrophin receptor antibody characteristics in a woman with long‐standing Hashimoto's who developed Graves' disease and pretibial myxoedema
Authors: Kamath C. , Young S. , Kabelis K. , Sanders J. , Adlan M. A. , Furmaniak J. , Rees Smith B. , Premawardhana L. D.
Published: April 2012

Context

Sequential conversion of Hashimoto's thyroiditis (HT) to Graves' disease (GD) is uncommon. Distinct immune paradigms, paucity of functioning tissue in long‐standing HT, and infrequent conversion of blocking (TBAb) to stimulating (TSAb) thyrotrophin receptor antibody (TRAb) may account for this. Molecular and crystal structure analysis helps delineate TSH receptor (TSHR)/TRAb interactions in detail. Such ‘fingerprinting’ helps determine the behaviour and characteristics of TRAb in longitudinal studies.

Patient

An 80‐year‐old woman taking thyroxine for long‐standing HT became hyperthyroid. This persisted despite thyroxine withdrawal – free T3 was 7·3 pmol/l (2·6–5·7) and TSH < 0·01 mU/l (0·2–4·5) and TRAb highly positive. She had a goitre (ultrasound – HT), pretibial myxoedema, with mild inactive Graves' orbitopathy. She had RAI treatment and is on thyroxine replacement.

Measurements and Results

Blood samples at presentation (A) and 1 year (B) showed high TSAb and TPOAb activity but no TBAb. Experiments involving TSHR mutations confirmed that (i) TRAb had stable characteristics over 1 year; (ii) TSHR mutation R255D caused complete inhibition and (iii) R109A caused marked reduction of cAMP production by M22 (TSHR‐stimulating human monoclonal antibody) and A and B; (iv) mutations R80A, E107A and K129A while affecting M22 had little effect on A and B.

Conclusions

The reasons for an immunological paradigm shift in this elderly woman remain speculative. We believe that de‐novo TSAb synthesis occurred converting her long‐standing HT to GD although the mechanisms responsible remain unexplained. TRAb analysis confirmed stable autoantibody characteristics over 1 year and variable effects of TSHR mutations on TRAb and M22 function.
Climate change and health with an emphasis on interactions with ultraviolet radiation: a review
Authors: Thomas Peter , Swaminathan Ashwin , Lucas Robyn M.
Published: April 2012

Climate change is increasingly recognized as a major risk to human health, and health concerns are assuming more importance in international debates on mitigation and adaptation strategies. Health consequences of climate change will occur through direct and indirect routes, and as a result of interactions with other environmental exposures. Heatwaves will become more common and are associated with higher mortality particularly in the elderly and those with pre‐existing cardiovascular and respiratory illnesses. Warmer ambient temperatures will result in more dehydration episodes and increased risks of renal disease and, through effects on pollen seasons, there may be an increase in allergic disease such as asthma and hayfever. Other adverse effects including on air quality, food safety and security and an expanding distribution of some infectious diseases, including vector‐borne diseases, are postulated. A related but separate environmental exposure is that of ultraviolet radiation (UVR). Interactions between climate change and stratospheric ozone (and the causes of ozone depletion) will cause changes to levels of ambient UVR in the future and warmer temperatures are likely to change sun exposure behaviour. Co‐occurring effects on aquatic and terrestrial ecosystems have potential consequences for food safety, quality and supply. Climate change‐related exposures are likely to affect the incidence and distribution of diseases usually considered as caused by UVR exposure; and changes in UVR exposure will modulate the climate change effects on human health. For example, in some regions warmer temperatures due to climate change will encourage more outdoor behaviour, with likely consequences for increasing skin cancer incidence. Although many of the health outcomes of both climate change and the interaction of climate change and UVR exposure are somewhat speculative, there are risks to over‐ or under‐estimations of health risks if synergistic and antagonistic effects of co‐occurring environmental changes are not considered.
Satellite‐based evidence for shrub and graminoid tundra expansion in northern Quebec from 1986 to 2010
Authors: McManus kelly M. , Morton Douglas C. , Masek Jeffrey G. , Wang Dongdong , Sexton Joseph O. , Nagol Jyoteshwar R. , Ropars Pascale , Boudreau Stéphane
Published: April 2012

Global vegetation models predict rapid poleward migration of tundra and boreal forest vegetation in response to climate warming. Local plot and air‐photo studies have documented recent changes in high‐latitude vegetation composition and structure, consistent with warming trends. To bridge these two scales of inference, we analyzed a 24‐year (1986–2010) Landsat time series in a latitudinal transect across the boreal forest‐tundra biome boundary in northern Quebec province, Canada. This region has experienced rapid warming during both winter and summer months during the last 40 years. Using a per‐pixel (30 m) trend analysis, 30% of the observable (cloud‐free) land area experienced a significant (P < 0.05) positive trend in the Normalized Difference Vegetation Index (NDVI). However, greening trends were not evenly split among cover types. Low shrub and graminoid tundra contributed preferentially to the greening trend, while forested areas were less likely to show significant trends in NDVI. These trends reflect increasing leaf area, rather than an increase in growing season length, because Landsat data were restricted to peak‐summer conditions. The average NDVI trend (0.007 yr−1) corresponds to a leaf‐area index (LAI) increase of ~0.6 based on the regional relationship between LAI and NDVI from the Moderate Resolution Spectroradiometer. Across the entire transect, the area‐averaged LAI increase was ~0.2 during 1986–2010. A higher area‐averaged LAI change (~0.3) within the shrub‐tundra portion of the transect represents a 20–60% relative increase in LAI during the last two decades. Our Landsat‐based analysis subdivides the overall high‐latitude greening trend into changes in peak‐summer greenness by cover type. Different responses within and among shrub, graminoid, and tree‐dominated cover types in this study indicate important fine‐scale heterogeneity in vegetation growth. Although our findings are consistent with community shifts in low‐biomass vegetation types over multi‐decadal time scales, the response in tundra and forest ecosystems to recent warming was not uniform.
The senses in practice: enhancing the quality of care for residents with dementia in care homes
Authors: Brown Wilson Christine , Swarbrick Caroline , Pilling Mark , Keady John
Published: April 2012

Aim.  The study aimed to develop, deliver, and evaluate a training programme in care homes to enhance the quality of care for people living with dementia based on the principles of relationship‐centred care expressed through the Senses Framework.

Background.  There are increasing numbers of people living with dementia worldwide with a growing proportion requiring residential long‐term care. This makes the quest for enhancing the quality of care and quality of life for people with dementia ever more pressing.

Design.  A mixed‐methods design was used adopting a Practice Development approach. The findings from one care home in the North West of England are reported.

Methods.  Eight facilitated workshops based on the principles of relationship‐centred care were completed and evaluated in 2010, using pre‐ and postintervention design. A focus group was undertaken with staff on completion of the study to gain a more comprehensive understanding of the practice/training context, augmented by case examples of changes in practice identified from the study workshops.

Results.  Structured questionnaires were used to profile the care home before and after the training. Following the workshops, staff felt more able to collect and use biographical information. In particular, staff reported how this information supported them to initiate meaningful conversations with the person with dementia as part of everyday care routines, thus improving overall feelings of well‐being.

Conclusion.  Using a biographical approach to care planning structured through the Senses Framework helped staff to develop a greater understanding of the person with dementia.