bluuebunny/crossref_metadata_2025 huggingface dataset:
Phosphoinositide [PI(3,5)P2] lipid-dependent regulation of the general transcriptional regulator Tup1
Authors: Han Bong-Kwan , Emr Scott D.
Published: May 2011

Transcriptional activity of a gene is governed by transcriptional regulatory complexes that assemble/disassemble on the gene and control the chromatin architecture. How cytoplasmic components influence the assembly/disassembly of transcriptional regulatory complexes is poorly understood. Here we report that the budding yeast Saccharomyces cerevisiae has a chromatin architecture-modulating mechanism that is dependent on the endosomal lipid PI(3,5)P2. We identified Tup1 and Cti6 as new, highly specific PI(3,5)P2 interactors. Tup1—which associates with multiple transcriptional regulators, including the HDAC (histone deacetylase) and SAGA complexes—plays a crucial role in determining an activated or repressed chromatin state of numerous genes, including GAL1. We show that, in the context that the Gal4 activation pathway is compromised, PI(3,5)P2 plays an essential role in converting the Tup1-driven repressed chromatin structure into a SAGA-containing activated chromatin structure at the GAL1 promoter. Biochemical and cell biological experiments suggest that PI(3,5)P2 recruits Cti6 and the Cyc8–Tup1 corepressor complex to the late endosomal/vacuolar membrane and mediates the assembly of a Cti6–Cyc8–Tup1 coactivator complex that functions to recruit the SAGA complex to the GAL1 promoter. Our findings provide important insights toward understanding how the chromatin architecture and epigenetic status of a gene are regulated by cytoplasmic components.
MDC1 directs chromosome-wide silencing of the sex chromosomes in male germ cells
Authors: Ichijima Yosuke , Ichijima Misako , Lou Zhenkun , Nussenzweig André , Camerini-Otero R. Daniel , Chen Junjie , Andreassen Paul R. , Namekawa Satoshi H.
Published: May 2011

Chromosome-wide inactivation is an epigenetic signature of sex chromosomes. The mechanism by which the chromosome-wide domain is recognized and gene silencing is induced remains unclear. Here we identify an essential mechanism underlying the recognition of the chromosome-wide domain in the male germline. We show that mediator of DNA damage checkpoint 1 (MDC1), a binding partner of phosphorylated histone H2AX (γH2AX), defines the chromosome-wide domain, initiates meiotic sex chromosome inactivation (MSCI), and leads to XY body formation. Importantly, MSCI consists of two genetically separable steps. The first step is the MDC1-independent recognition of the unsynapsed axis by DNA damage response (DDR) factors such as ataxia telangiectasia and Rad3-related (ATR), TOPBP1, and γH2AX. The second step is the MDC1-dependent chromosome-wide spreading of DDR factors to the entire chromatin. Furthermore, we demonstrate that, in somatic cells, MDC1-dependent amplification of the γH2AX signal occurs following replicative stress and is associated with transcriptional silencing. We propose that a common DDR pathway underlies both MSCI and the response of somatic cells to replicative stress. These results establish that the DDR pathway centered on MDC1 triggers epigenetic silencing of sex chromosomes in germ cells.
The RNA polymerase II C-terminal domain promotes splicing activation through recruitment of a U2AF65–Prp19 complex
Authors: David Charles J. , Boyne Alex R. , Millhouse Scott R. , Manley James L.
Published: May 2011

Pre-mRNA splicing is frequently coupled to transcription by RNA polymerase II (RNAPII). This coupling requires the C-terminal domain of the RNAPII largest subunit (CTD), although the underlying mechanism is poorly understood. Using a biochemical complementation assay, we previously identified an activity that stimulates CTD-dependent splicing in vitro. We purified this activity and found that it consists of a complex of two well-known splicing factors: U2AF65 and the Prp19 complex (PRP19C). We provide evidence that both U2AF65 and PRP19C are required for CTD-dependent splicing activation, that U2AF65 and PRP19C interact both in vitro and in vivo, and that this interaction is required for activation of splicing. Providing the link to the CTD, we show that U2AF65 binds directly to the phosphorylated CTD, and that this interaction results in increased recruitment of U2AF65 and PRP19C to the pre-mRNA. Our results not only provide a mechanism by which the CTD enhances splicing, but also describe unexpected interactions important for splicing and its coupling to transcription.
Sweet DREAMs for Hippo
Authors: Dick Frederick A. , Mymryk Joe S.
Published: May 2011

The Hippo pathway coordinates organ size and cell proliferation. The retinoblastoma family of proteins regulates progression through the G0/G1 phase of the cell cycle. Disruption of either pathway contributes to cancer formation. Three recent studies in Genes & Development reveal how cellular proliferation is coordinated between these pathways. Here we discuss the implications of these studies and the new questions that they raise.
Of blood, bones, and ribosomes: is Swachman-Diamond syndrome a ribosomopathy?
Authors: Johnson Arlen W. , Ellis Steve R.
Published: May 2011

Mutations in the human SBDS (Shwachman-Bodian-Diamond syndrome) gene are the most common cause of Shwachman-Diamond syndrome, an inherited bone marrow failure syndrome. In this issue of Genes & Development, Finch and colleagues (pp. 917–929) establish that SBDS functions in ribosome synthesis by promoting the recycling of eukaryotic initiation factor 6 (eIF6) in a GTP-dependent manner. This work supports the idea that a ribosomopathy may underlie this syndrome.
Addicted to Myc—but why?: Figure 1.
Authors: von Eyss Björn , Eilers Martin
Published: May 2011

Many tumors continuously depend on the initiating oncogenes, but whether this extends to their downstream targets is unclear. In this issue of Genes & Development, Sodir and colleagues (pp. 907–916) demonstrate an essential role for endogenous Myc proteins in maintaining the tumor microenvironment, providing an unexpected molecular explanation for addiction to Myc.
Nuclear actin polymerization is required for transcriptional reprogramming of Oct4 by oocytes
Authors: Miyamoto Kei , Pasque Vincent , Jullien Jerome , Gurdon John B.
Published: May 2011

Amphibian oocytes can rapidly and efficiently reprogram the transcription of transplanted somatic nuclei. To explore the factors and mechanisms involved, we focused on nuclear actin, an especially abundant component of the oocyte's nucleus (the germinal vesicle). The existence and significance of nuclear actin has long been debated. Here, we found that nuclear actin polymerization plays an essential part in the transcriptional reactivation of the pluripotency gene Oct4 (also known as Pou5f1). We also found that an actin signaling protein, Toca-1, enhances Oct4 reactivation by regulating nuclear actin polymerization. Toca-1 overexpression has an effect on the chromatin state of transplanted nuclei, including the enhanced binding of nuclear actin to gene regulatory regions. This is the first report showing that naturally stored actin in an oocyte nucleus helps transcriptional reprogramming in a polymerization-dependent manner.
A novel function of the proneural factor Ascl1 in progenitor proliferation identified by genome-wide characterization of its targets
Authors: Castro Diogo S. , Martynoga Ben , Parras Carlos , Ramesh Vidya , Pacary Emilie , Johnston Caroline , Drechsel Daniela , Lebel-Potter Mélanie , Garcia Laura Galinanes , Hunt Charles , Dolle Dirk , Bithell Angela , Ettwiller Laurence , Buckley Noel , Guillemot François
Published: May 2011

Proneural genes such as Ascl1 are known to promote cell cycle exit and neuronal differentiation when expressed in neural progenitor cells. The mechanisms by which proneural genes activate neurogenesis—and, in particular, the genes that they regulate—however, are mostly unknown. We performed a genome-wide characterization of the transcriptional targets of Ascl1 in the embryonic brain and in neural stem cell cultures by location analysis and expression profiling of embryos overexpressing or mutant for Ascl1. The wide range of molecular and cellular functions represented among these targets suggests that Ascl1 directly controls the specification of neural progenitors as well as the later steps of neuronal differentiation and neurite outgrowth. Surprisingly, Ascl1 also regulates the expression of a large number of genes involved in cell cycle progression, including canonical cell cycle regulators and oncogenic transcription factors. Mutational analysis in the embryonic brain and manipulation of Ascl1 activity in neural stem cell cultures revealed that Ascl1 is indeed required for normal proliferation of neural progenitors. This study identified a novel and unexpected activity of the proneural gene Ascl1, and revealed a direct molecular link between the phase of expansion of neural progenitors and the subsequent phases of cell cycle exit and neuronal differentiation.
Isolation of Human Capillary Endothelial Cells Using Paramagnetic Beads Conjugated to Anti-PECAM Antibodies
Authors: Springhorn Jeremy P.
Published: May 2011

INTRODUCTION

The protocol described below outlines a technique to isolate and maintain pure populations of human capillary endothelial cells using an active selection technique that targets the platelet endothelial cell adhesion molecule (PECAM or CD31). Human abdominal wall adipose tissue is used as the source for capillary endothelial cells, which are partially purified before selection.
Drosophila Appetitive Olfactory Conditioning: Figure 1.
Authors: Krashes Michael J. , Waddell Scott
Published: May 2011

INTRODUCTION

Olfactory memory paradigms have been extensively used to study memory in Drosophila since the late 1960s, and over the intervening years, investigators have made a series of worthwhile changes and “tweaks” to the procedures. This protocol provides the reader with a detailed description of the appetitive (odor–sugar) memory assay currently used in the Waddell laboratory. We also describe the essential control assays for sensory acuity and locomotor behavior. It should be emphasized that the assays we describe work adequately in our laboratory, but are also amenable to, and may be further improved by additional adjustments.
Conjugation of Magnetic Beads for Immunopurification of Protein Complexes
Authors: Cristea Ileana M. , Chait Brian T.
Published: May 2011

INTRODUCTION

Knowledge of the composition of protein complexes provides key insights into their functions. Immunoaffinity purification provides an effective means for isolating protein complexes and elucidating their composition. Immunoisolation is achieved with antibodies directed either specifically against the proteins of interest or against tags that are coupled to the proteins of interest. This approach uses immunoaffinity purification on magnetic beads coated with antibodies for the rapid and efficient purification of protein complexes from cells or tissues. This protocol describes conjugation of magnetic beads with antibodies in preparation for immunopreciptation of protein complexes.
Dissection of Embryonic Mouse Kidney, Culture In Vitro, and Imaging of the Developing Organ
Authors: Costantini Frank , Watanabe Tomoko , Lu Benson , Chi Xuan , Srinivas Shankar
Published: May 2011

INTRODUCTION

In this article, we outline procedures for the dissection of intact kidneys and the isolation of the ureteric bud (UB) and the metanephric mesenchyme (MM) from mouse embryos. The apparatus required for the culture of these tissues in vitro is described in detail as well as the equipment necessary for performing time-lapse imaging studies of the developing kidney.
Characterization of Protein–DNA Interactions Using Protein Microarrays
Authors: Hu Shaohui , Xie Zhi , Blackshaw Seth , Qian Jiang , Zhu Heng
Published: May 2011

INTRODUCTION

Protein–DNA interactions (PDIs) are critical for many cellular processes. We present here a protocol for the identification of PDIs in vitro using protein microarray technology. The procedure involves double-stranding synthesized DNA oligonucleotides with a fluorescent-labeled primer, binding the labeled double-stranded DNA directly to the protein microarray, and analyzing binding of the resulting PDIs. This approach provides simultaneous identification of PDIs for thousands of proteins, and multiple carefully designed DNA probes can be tested in parallel, which enables a rapid mapping of PDIs on a proteome-wide scale.
Generation of Replication-Competent and -Defective HSV Vectors
Authors: Goins William F. , Krisky David M. , Wechuck James B. , Wolfe Darren , Huang Shaohua , Glorioso Joseph C.
Published: May 2011

INTRODUCTION

Engineering effective vectors has been crucial to the efficient delivery and expression of therapeutic gene products in vivo. Among these, HSV-1 represents an excellent candidate vector for delivery to the peripheral and central nervous systems. The natural biology of HSV-1 includes the establishment of a lifelong latent state in neurons in which the viral genome persists as an episomal molecule. Genomic HSV vectors can be produced that are completely replication-defective, nontoxic, and capable of long-term transgene expression. Herpes simplex virus (HSV) vectors are constructed by using a replication-deficient vector backbone (TOZ.1) for homologous recombination with a shuttle plasmid containing a cassette expressing the gene of interest inserted into the UL41 gene sequence. The TOZ.1 vector expresses a reporter gene (lacZ) in the UL41 locus, such that recombination of the transgenic cassette into the UL41 locus results in the loss of the reporter gene activity. The TOZ.1 vector also contains a unique PacI endonuclease site for digestion of parental viral DNA that substantially reduces the nonrecombinant background. Following homologous recombination of the shuttle plasmid into the PacI-digested TOZ.1 genome, the recombinants are identified as clear plaques. After three rounds of limiting dilution analysis, the structure of the recombinants can be confirmed by Southern blot or by polymerase chain reaction (PCR) analysis.
Two-Photon Uncaging Microscopy
Authors: Matsuzaki Masanori , Kasai Haruo
Published: May 2011

INTRODUCTION

Two-photon uncaging takes advantage of the inherent optical sectioning power of two-photon excitation to generate highly localized concentration increases of neurotransmitters such as glutamate. This can be used to activate isolated clusters of receptors and, thus, produce maps of receptor densities, or activate intracellular signal transduction under these receptors, in three dimensions and in complex structures such as hippocampal brain slices. Used in combination with two-photon imaging, two-photon uncaging provides a means to study the long-term structural and functional consequences of stimulation of structures such as dendritic spines. This protocol gives an overview of the procedures used for two-photon uncaging microscopy. It includes a detailed description of the development of a microscope that enables effective two-photon release of caged neurotransmitters and provides several examples of its use in cultured and acutely isolated hippocampal neurons.
Labeling Membrane Glycoproteins or Glycolipids with Fluorescent Wheat Germ Agglutinin
Authors: Chazotte Brad
Published: May 2011

INTRODUCTION

Glycoprotein and glycolipids are found as part of the outer leaflet of cellular plasma membranes. Those glycoproteins or glycolipids that contain sialic acid and N-acetylglucosamine residues can be labeled with wheat germ agglutinin (WGA), a plant lectin that exists as a dimer (mwt ∼36,000) and is normally cationic. Fluorescently labeled WGA is commercially available with fluorescein, Alexa Fluor, or rhodamine moieties. Fluorescent WGA can also be used to stain the Golgi (the trans-Golgi) in fixed cells. Fluorescently labeled concanavalin A (Con A), which selectively binds to α-mannopyranosyl and α-glucopyranosyl residues, can also be used to label plasma membranes. This protocol describes the labeling of membrane glycoproteins or glycolipids with fluorescent WGA.
Labeling Membranes with Fluorescent Cholesterols
Authors: Chazotte Brad
Published: May 2011

INTRODUCTION

Cholesterol, an essential component of most cellular membranes, is present in the plasma membrane and is enriched within lipid rafts. It is, however, virtually absent from the mitochondrial inner membrane, and there is little within the endoplasmic reticulum. Dehydroergosterol (DHE), a fluorescent cholesterol analog, has been found to effectively label membranes and has been used to monitor cholesterol distribution and trafficking within live cells. Other cholesterol-based probes, such as NBD (N-(7-nitrobenz-2-oxa-1,3-diazole-4-yl))-cholesterol, are commercially available (from Molecular Probes). However, cell labeling using the NBD-cholesterol analogs frequently does not show the same labeling pattern as for DHE and native cholesterol.
Fluorescent Labeling of Membrane Lipid Rafts
Authors: Chazotte Brad
Published: May 2011

INTRODUCTION

Lipid rafts are structures within the plasma membrane that have a different lipid composition than the bulk membrane, specifically being enriched in certain lipids. Lipid rafts are believed to have physiologically important functions. Commercially available kits have been developed (Invitrogen/Molecular Probes) for labeling lipid rafts using a cholera toxin subunit that has been fluorescently tagged with one of several Alexa Fluor dyes. As described in this protocol, cells are labeled while in suspension, after which they can be mounted for imaging.
Pivotal role of epithelial cell adhesion molecule in the survival of lung cancer cells
Authors: Hase Tetsunari , Sato Mitsuo , Yoshida Kenya , Girard Luc , Takeyama Yoshihiro , Horio Mihoko , Elshazley Momen , Oguri Tomoyo , Sekido Yoshitaka , Shames David S. , Gazdar Adi F. , Minna John D. , Kondo Masashi , Hasegawa Yoshinori
Published: May 2011

Epithelial cell adhesion molecule (EpCAM) is overexpressed in a wide variety of human cancers including lung cancer, and its contribution to increased proliferation through upregulation of cell cycle accelerators such as cyclins A and E has been well established in breast and gastric cancers. Nevertheless, very little is known about its role in supporting the survival of cancer cells. In addition, the functional role of EpCAM in the pathogenesis of lung cancer remains to be explored. In this study, we show that RNAi‐mediated knockdown of EpCAM suppresses proliferation and clonogenic growth of three EpCAM‐expressing lung cancer cell lines (H3255, H358, and HCC827), but does not induce cell cycle arrest in any of these. In addition, EpCAM knockdown inhibits invasion in the highly invasive H358 but not in less invasive H3255 cells in a Transwell assay. Of note, the EpCAM knockdown induces massive apoptosis in the three cell lines as well as in another EpCAM‐expressing lung cancer cell line, HCC2279, but to a much lesser extent in a cdk4/hTERT immortalized normal human bronchial epithelial cell line, HBEC4, suggesting that EpCAM could be a therapeutic target for lung cancer. Finally, EpCAM knockdown partially restores contact inhibition in HCC827, in association with p27Kip1 upregulation. These results indicate that EpCAM could contribute substantially to the pathogenesis of lung cancer, especially cancer cell survival, and suggest that EpCAM targeted therapy for lung cancer may have potential. (Cancer Sci 2011; 102: 1493–1500)
Myeloperoxidase is associated with incident coronary heart disease independently of traditional risk factors: results from the MONICA/KORA Augsburg study
Authors: Karakas M. , Koenig W. , Zierer A. , Herder C. , Rottbauer W. , Baumert J. , Meisinger C. , Thorand B.
Published: May 2011

Abstract.  Karakas M, Koenig W, Zierer A, Herder C, Rottbauer W, Baumert J, Meisinger C, Thorand B (University of Ulm Medical Center, Ulm; German Research Center for Environmental Health, Institute of Epidemiology II, Neuherberg; and Institute for Clinical Diabetology, Leibniz Center for Diabetes Research at Heinrich Heine University, Düsseldorf, Germany). Myeloperoxidase is associated with incident coronary heart disease independently of traditional risk factors: results from the MONICA/KORA Augsburg study. J Intern Med 2012; 271: 43–50.

Aims.  Oxidative stress plays a critical role in the initiation and progression of atherosclerosis. Myeloperoxidase (MPO) is a marker of oxidative stress. We prospectively investigated whether an increased serum concentration of MPO is associated with an increased risk of incident coronary heart disease (CHD).

Methods.  We conducted a population‐based case‐cohort study in middle‐aged, healthy men and women within the MONICA/KORA Augsburg studies. Serum levels of MPO were measured in 333 subjects with (cases) and 1727 without (noncases) incident CHD. Mean follow‐up time was 10.8 ± 4.6 years.

Results.  Baseline concentrations of MPO were higher in cases compared with noncases (P ≤ 0.001 in men; P = 0.131 in women). After adjustment for major cardiovascular risk factors, the hazard ratio (HR) with 95% confidence interval (CI) comparing the top with the two lower tertiles was 1.70 (95% CI, 1.25–2.30). After additional adjustment for markers of inflammation and endothelial dysfunction, the association was attenuated (HR 1.50; 95% CI, 1.08–2.09). There were no significant interactions of MPO with sex or increased weight on CHD risk.

Conclusions.  Elevated concentrations of the oxidative stress marker MPO were independently associated with increased risk of incident CHD. This finding deserves detailed evaluation in further studies.
Pepper asparagine synthetase 1 (CaAS1) is required for plant nitrogen assimilation and defense responses to microbial pathogens
Authors: Hwang In Sun , An Soo Hyun , Hwang Byung Kook
Published: May 2011

Asparagine synthetase is a key enzyme in the production of the nitrogen‐rich amino acid asparagine, which is crucial to primary nitrogen metabolism. Despite its importance physiologically, the roles that asparagine synthetase plays during plant defense responses remain unknown. Here, we determined that pepper (Capsicum annuum) asparagine synthetase 1 (CaAS1) is essential for plant defense to microbial pathogens. Infection with Xanthomonas campestris pv. vesicatoria (Xcv) induced early and strong CaAS1 expression in pepper leaves and silencing of this gene resulted in enhanced susceptibility to Xcv infection. Transgenic Arabidopsis (Arabidopsis thaliana) plants that overexpressed CaAS1 exhibited enhanced resistance to Pseudomonas syringae pv. tomato DC3000 and Hyaloperonospora arabidopsidis. Increased CaAS1 expression influenced early defense responses in diseased leaves, including increased electrolyte leakage, reactive oxygen species and nitric oxide bursts. In plants, increased conversion of aspartate to asparagine appears to be associated with enhanced resistance to bacterial and oomycete pathogens. In CaAS1‐silenced pepper and/or CaAS1‐overexpressing Arabidopsis, CaAS1‐dependent changes in asparagine levels correlated with increased susceptibility or defense responses to microbial pathogens, respectively. Linking transcriptional and targeted metabolite studies, our results suggest that CaAS1 is required for asparagine synthesis and disease resistance in plants.
Cutaneous epitheliotropic T‐cell lymphoma in the cat: a review of the literature and five new cases
Authors: Fontaine Jacques , Heimann Marianne , Day Michael J.
Published: May 2011

Cutaneous epitheliotropic T‐cell lymphoma (CETL) is characterized by cutaneous infiltration of neoplastic T lymphocytes with a specific tropism for the epidermis and adnexal epithelium. This disease is reported very rarely in the cat. Clinical data were collected from an informal discussion with veterinary dermatologists through the Vetdermlist (vetderm@lists.ncsu.edu). In parallel, case archives of two European diagnostic histopathology laboratories (Institut de Pathologie et Génétique/Bio.be Gosselies, Belgium and the School of Veterinary Sciences, University of Bristol, UK) were reviewed. Fifteen cases with a good clinical description were selected, and five sets of skin biopsies were available for review. Cutaneous epitheliotropic T‐cell lymphoma generally affects older cats with no apparent sex or breed predisposition. Solitary or multiple lesions were reported without predilection for any particular location. The lesions consisted of erythematous plaques or patches, scaly alopecic patches and nonhealing ulcers or nodules, which sometimes mimicked an eosinophilic plaque. Pruritus was rarely reported. No lesions affecting the oral mucosa were observed. Clinical diagnosis of CETL is more challenging in cats than in dogs. Final diagnosis must be based on histopathological examination of skin biopsy samples. The characteristic lesions of feline CETL are similar to those reported in the dog, but involvement of the adnexal glands was not observed in this series (n = 5). The neoplastic T cells were generally small to medium in size. The survival time of cats with CETL seems to be more variable than that of affected dogs. Too few cases have been evaluated to permit clear recommendations to be made with respect to treatment.
Microchip‐associated fibrosarcoma in a cat
Authors: Carminato Antonio , Vascellari Marta , Marchioro Wendy , Melchiotti Erica , Mutinelli Franco
Published: May 2011

A 9‐year‐old, neutered male cat was presented for a subcutaneous mass on the neck. After surgical removal of the mass, a pet identification microchip was found within the tumour. Histological examination of the mass revealed typical features of the feline postinjection sarcoma. The cat had never received injections at the tumour site; all routine vaccinations were administered in the hindlimbs. Few cases of sarcomas developing at the site of microchip application have been reported in animals, although the contributory role of vaccine administrations has not been ruled out. This is the first report of a microchip‐associated fibrosarcoma in a cat. Adherence to American Association of Feline Practitioners vaccination guidelines, avoiding the interscapular area, enabled confirmation of the definitive aetiology of the neoplasia.
The effects of skin disease on the penetration kinetics of hydrocortisone through canine skinin vitro
Authors: Ahlstrom Liisa A. , Cross Sheree E. , Mills Paul C.
Published: May 2011

This study investigated the effects of allergic skin disease on the penetration kinetics of hydrocortisone through canine skinin vitro. Full‐thickness lesional and nonlesional (normal) skin was removed from the dorsal lumbosacral and dorsocaudal thoracic regions, respectively, of five canine cadavers. The dogs were suspected of having flea allergy dermatitis based on their distribution and types of skin lesions. Nonlesional skin was confirmed to be histologically normal, and the histopathology of the lesional skin was consistent with allergic dermatitis. Excised skin was clipped, mounted in Franz‐type diffusion cells, and the transdermal penetration of a saturated, radiolabelled hydrocortisone solution was measured over 30 h. When the penetration data for all five dogs were pooled, a restricted (or residual) maximal likelihood mixed model predicted that the permeability coefficient and pseudosteady‐state flux of hydrocortisone was more than twice as great (95% confidence interval 1.55–2.71 times as great;P < 0.0001) through lesional compared with nonlesional skin. There was no significant difference in the lag time for hydrocortisone penetration through lesional compared with nonlesional skin of the dogs. This study has confirmed that the transdermal penetration of hydrocortisone may be altered, typically increased twofold, but could be as high as 10‐fold, through lesional compared with nonlesional skin of dogs with suspected flea allergy dermatitis. This is likely to be affected by variables such as disease severity, concurrent infections and interindividual differences in skin characteristics.
Evaluation of the bacterial microflora of the conjunctival sac of healthy dogs and dogs with atopic dermatitis
Authors: Furiani Nicla , Scarampella Fabia , Anna Martino Piera , Panzini Ilaria , Fabbri Elisabetta , Ordeix Laura
Published: May 2011

The aim of this case–control study was to evaluate and compare the bacterial microflora from the conjunctival sac of dogs with atopic dermatitis and healthy dogs. Twenty‐one atopic dogs without clinical and/or cytopathological signs of bacterial blepharoconjunctivitis and 21 breed‐matched healthy dogs were enrolled. Under topical anaesthesia, the inferior conjunctival sac of one eye was scraped twice. Material was collected with a Kimura spatula, spread over a slide and stained with a Diff Quick®‐type stain (Medion Diagnostics GmbH, Düdingen, Switzerland) for cytological examination. An area of 0.5 cm2 was examined at ×1000 magnification, and the types and numbers of cells and bacteria were recorded. A bacterial swab was collected and inoculated into culture media for the growth of aerobic bacteria. Before sampling, each atopic dog was evaluated for severity of cutaneous lesions, pruritus and conjunctival inflammation. Significant differences were observed between atopic and healthy dogs for the presence of bacteria on cytology (P = 0.015), keratinized (P = 0.001) and nonkeratinized epithelial cells (P = 0.013), eosinophils (P = 0.019) and lymphocytes (P = 0.008). Bacteria were recovered from 12 atopic dogs and three healthy dogs (P = 0.004). Staphylococcus pseudintermedius was the most commonly isolated species in atopic dogs (seven of 12). In atopic dogs, no significant relation was found between conjunctival bacterial colonization (on cytology and culture) and the severity of any of the clinical parameters. This study suggests differences in conjunctival bacterial colonization and cytological features between atopic and healthy dogs.
Epithelial migration on the canine tympanic membrane
Authors: Tabacca Natalie E. , Cole Lynette K. , Hillier Andrew , Rajala‐Schultz Päivi J.
Published: May 2011

Epithelial migration (EM) is a process that serves as a self‐cleaning and repair mechanism for the ear canal and tympanic membrane (TM). Epithelial migration has been evaluated in humans and several other species, but not in dogs. The objective of this study was to determine the rate and pattern of EM on the TM in clinically normal laboratory dogs. Eighteen dogs were anaesthetized, and three drops of waterproof drawing ink were placed on two sites of the pars tensa (PT) and one on the pars flaccida (PF). Images were recorded with a video otoscope and digital capture system. Each dog was evaluated and images were recorded every 6–8 days for four evaluations. Migration pattern analysis and EM rate calculation were performed with image‐processing software. Descriptive statistics for EM rate (mean, SD, 95% confidence interval) were calculated for all ink‐drop locations on the TM (PT1, PT2 and PF) at each time point. No significant differences in the mean EM rates were identified between right and left ears of the fox hounds breeds (beagle or fox hound), or locations PT1 and PT2. The mean overall EM rates (±SD) were 96.4 (±43.1) and 225.4 (±128.1) μm/day for the PT and PF, respectively. All ink drops moved outwards, the majority in a radial direction, from the original location to the periphery of the TM. The ink‐drop placement method used in this study can be used in future studies to determine the EM rate of the canine TM.
Effects of species and season on chemical composition and ruminal crude protein and organic matter degradability of some multi‐purpose tree species by West African dwarf rams
Authors: Arigbede O. M. , Anele U. Y. , Südekum K.‐H. , Hummel J. , Oni A. O. , Olanite J. A. , Isah A. O.
Published: May 2011

Seasonal chemical composition and ruminal organic matter (OM) and crude protein (CP) degradabilities were determined in four tropical multi‐purpose tree species (MPTS) namely; Pterocarpus santalinoides, Grewia pubescens, Enterolobium cyclocarpum and Leucaena leucocephala. Three West African dwarf (WAD) rams fitted with permanent rumen cannula were used for the degradability trials. Foliage samples were collected four times to represent seasonal variations as follows: January – mid dry; April – late dry; July – mid rainy and October – late rainy seasons. Leaf samples were randomly collected from the trees for estimation of dry matter (DM) and chemical composition. Ruminal in sacco OM and CP degradabilities were estimated from residues in nylon bags. All samples had high CP (161–259 g/kg DM) and moderate fibre concentrations [neutral detergent fibre (without residual ash], 300–501 g/kg DM; acid detergent fibre (without residual ash), 225–409 g/kg DM and acid detergent lignin, 87–179 g/kg DM across seasons. Interaction effects of species and season on chemical composition were highly significant (p = 0.001) except for trypsin inhibitor (p = 0.614). The MPTS recorded more than 60% OM and CP degradability at 24 h, which implied that they were all highly degradable in the rumen. Their incorporation into ruminant feeding systems as dry season forage supplements is therefore recommended.
Improved heterologous protein expression in the chloroplast of Chlamydomonas reinhardtii through promoter and 5′ untranslated region optimization
Authors: Rasala Beth A. , Muto Machiko , Sullivan Joseph , Mayfield Stephen P.
Published: May 2011

Microalgae have the potential to be a valuable biotechnological platform for the production of recombinant proteins. However, because of the complex regulatory network that tightly controls chloroplast gene expression, heterologous protein accumulation in a wild‐type, photosynthetic‐competent algal chloroplast remains low. High levels of heterologous protein accumulation have been achieved using the psbA promoter/5′ untranslated region (UTR), but only in a psbA‐deficient genetic background, because of psbA/D1‐dependent auto‐attenuation. Here, we examine the effect of fusing the strong 16S rRNA promoter to the 5′ UTR of the psbA and atpA genes on transgene expression in the chloroplast of Chlamydomonas reinhardtii. We show that fusion of the 16S promoter had little impact on protein accumulation from the psbA 5′ UTR in a psbA‐deficient genetic background. Furthermore, the 16S/psbA promoter/UTR fusion was silenced in the presence of wild‐type levels of D1 protein, confirming that the psbA 5′ UTR is the primary target for D1‐dependent auto‐repression. However, fusion of the 16S promoter to the atpA 5′ UTR significantly boosts mRNA levels and supports high levels of heterologous protein accumulation in photosynthetic‐competent cells. The 16S/atpA promoter/UTR drove LUXCT protein accumulation to levels close to that of psbA in a psbA− background, and drove expression of a human therapeutic protein to levels only twofold lower than the psbA 5′ UTR. The 16S/atpA promoter/UTR combination should have utility for heterologous protein production when expression from a photosynthetic‐competent microalgal strain is required.
The Influence of Spanish‐Language Media on Latino Public Opinion and Group Consciousness*
Authors: Kerevel Yann P.
Published: May 2011

Objective. This article determines if the use of Spanish‐language media among Latinos influences public opinion on various policy issues and group consciousness.

Methods. Using a 2004 national public opinion survey of U.S. Latinos, a multivariate analysis is run to determine the effect of language media preference on immigration policy, abortion, same‐sex marriage, and three measures of group consciousness.

Results. I find more frequent use of Spanish‐language media leads to more liberal attitudes toward immigration, but has no effect on opinions toward abortion and same‐sex marriage. I also find increased use of Spanish‐language media leads to increased levels of group consciousness.

Conclusions. The differences in attitudes are due to the diverging goals of Spanish‐language and English‐language media. The effect of using Spanish‐language media serves to promote a sense of group consciousness among Latinos by reinforcing roots in Latin America and the commonalities among Latinos of varying national origin.
Are Debates about “Morality Policy” Really about Morality? Framing Opposition to Gay and Lesbian Rights
Authors: Mucciaroni Gary
Published: May 2011

A growing number of studies identify “morality policy” as a distinct category of public policy and have tested several related hypotheses. This article reexamines morality policy as a conceptual category and an empirical phenomenon. As others have pointed out, we should distinguish morality policy from other policies by how political actors frame issues rather than by its substantive content. In the first part of the article, I argue that we should view morality “policy” as one of two broad strategies for framing issues, rather than try to fit it into existing policy typologies. Next, I move beyond viewing morality policy as a single, broad category by identifying several distinct subtypes of morality frames. In the second part of the article, I challenge a basic assumption of the morality policy paradigm—that advocates frame morality policy issues by engaging in moralistic discourse that reflects their basic beliefs and values. Gay rights issues are a strong test of this claim because the literature cites them as typical examples of morality policy, and gay rights opponents would seem especially likely to engage in “morality talk” in framing these issues. Very few studies of morality policy actually observe framing behavior and what it reveals about the political strategy of each side. Congressional and state‐level data reveal that opponents usually do not frame gay rights issues in terms of the morality of homosexuality or religious injunctions against it, even in most states where we would expect to find it. Instead, they emphasize frames that focus on alleged negative social consequences from gay rights and procedural arguments about who should make policy and how it should be made. Although many opponents of gay rights disapprove of homosexuality on moral and religious grounds, their framing behavior reflects more complex strategic considerations. I speculate that opponents deemphasize morality talk because it is politically disadvantageous compared with other kinds of frames, and because of greater acceptance of gays in society. In reducing gay rights debates to moral and religious judgments, the morality policy perspective obscures the complexity of advocates' framing strategies and ignores many of their most important arguments.
The Friends and Family Plan: Contact with Gays and Support for Gay Rights
Authors: Lewis Gregory B.
Published: May 2011

According to both the contact hypothesis and gay rights advocates, coming out to straight friends and family members should increase acceptance of homosexuality and support for gay rights. If lesbians, gay men, and bisexuals (LGBs) come out primarily to people they expect to be accepting, however, the repeated finding that people who know LGBs are more likely to support gay rights could be overstating the impact of coming out. Using individual‐level data from 27 national surveys, I find that similar variables predict both knowing LGBs and supporting gay rights, but in different ways. Even after controlling for those demographic, religious, and political variables—and sometimes also for beliefs about whether some people are born gay and whether homosexuality is immoral—people who know LGBs are much more likely to support gay rights. The effect holds for every issue, in every year, for every type of relationship, and for every demographic, religious, and political subgroup.
Constructing Homeland Security: An Anemic Policy Regime
Authors: May Peter J. , Jochim Ashley E. , Sapotichne Joshua
Published: May 2011

This research addresses the strength of the homeland security policy regime that was constructed after the terrorist attacks of September 2001. We argue that homeland security provides a preeminent example of the challenges of developing policy regimes that focus policymaking on a common goal across diverse subsystems. All the ingredients for fashioning a powerful regime were in place after the terrorist attacks of September 2001—a common purpose, engaged stakeholders, and institutional redesign. But for a variety of reasons that we discuss, the results are far from cohesive. The lessons we draw are more general ones regarding factors that influence the strength of boundary‐spanning policy regimes.
Deterrence Theory and the Implementation of Speed Limits in the American States
Authors: Ritchey Mark , Nicholson‐Crotty Sean
Published: May 2011

Despite the ubiquitous nature of these policies, there is disagreement in the literature regarding the direction and size of the impact that speed limits have on traffic‐related fatalities. We argue in this paper that the mixed results in previous work may arise because these studies have missed an important component of the implementation of speed limit laws. More explicitly, they have failed to adequately control for the deterrent effect of enforcement and sanctions. We develop the argument that the observed impacts of speed limits will be overly large when the certainty and severity of punishment are not accounted for. We test this assertion in a cross‐sectional time series analysis of state‐level traffic fatalities from the years 1990–2006 and find that lower speeds do save a significant number of lives. Interestingly, we find that the impact is significantly overestimated for 65‐mph limits and significantly underestimated for 70‐mph limits when enforcement, penalties, and the interaction of the two are excluded. The results also suggest that fines have a rather modest impact on fatalities unless states employ a sufficient number of troopers to enforce posted limits. In addition to clarifying previous findings related to speed limit policy, therefore, the findings contribute to the general application of deterrence theory by empirically confirming that the importance of sanction severity is dependent on the perceived certainty of punishment.
The Effect of Vacancy and Barium Substitution on the Stability of the Cesium Titanium Silicate Pollucite
Authors: Park Tae‐Jin , Garino Terry J. , Nenoff Tina M. , Rademacher David , Navrotsky Alexandra
Published: May 2011

Cesium titanium silicate (CsTiSi2O6.5) is a titanium analogue of pollucite CsAlSi2O6 and a possible ceramic form for immobilization of short‐lived fission products in radioactive waste. Through β decay, cesium (Cs) decays to barium. Therefore, not only the stability of Cs‐loaded waste forms, but also that of a potential decay product series is of fundamental importance. Ba‐substituted CsTiSi2O6.5 is a potential β decay product with the pollucite structure. Here, we report the effects of the reaction synthesis condition and the study of the thermodynamic stability of potential intermediates in the decay product series (1) with charge‐balance in pollucite as two Cs ions are replaced by one Ba and a vacancy and (2) with one‐to‐one replacement of Cs by Ba. The enthalpies of formation of Ba‐substituted CsTiSi2O6.5 were obtained from drop solution calorimetry in a molten lead borate solvent at 702°C. The enthalpies of formation, from constituent oxides, are exothermic and it decreases with increasing Ba content. The effect of vacancies in the pollucite structure is a more dominant factor in the energetics than that of Ba replacement. The thermodynamic effects of acetate and/or nitrate precursors and of adding acetic acid during synthesis of single phase Ba‐substituted CsTiSi2O6.5 pollucite are insignificant except for the sample prepared from acetate precursors without acid treatment.
Preparation and Characteristics of Bi0.5Na0.5TiO3 Single‐Crystalline Films by a Solid‐State Process
Authors: Ijuu Daiji , Kimura Toshio , Yamada Tomoaki , Funakubo Hiroshi
Published: May 2011

A novel process was demonstrated for the preparation of an epitaxial Bi0.5Na0.5TiO3 (BNT) single‐crystalline film on a SrTiO3 single crystal substrate by simply heating a composite consisting of a BNT powder layer on the substrate. The BNT powder layer was formed on the substrate by spin coating and the composite was heated at 1150°C for several hours in a lidded crucible containing the BNT powder to avoid evaporation of Na and Bi from the composite. This yielded an epitaxial BNT single‐crystalline film several micrometers thick. The film had a high degree of orientation with the rocking curve full width at half maximum of 0.064°. The mechanism of film formation was found to be solid‐state spreading of BNT powder particles on the substrate, and a dense film was obtained from the porous powder layer.
Novel Low‐Temperature Cofired Ceramic System Containing Plate‐like Filler
Authors: Cho Beom‐Joon , Lee Jong‐Myeon , Park Yun‐Hwi
Published: May 2011

A new LTCC (low‐temperature cofired ceramic) system containing Al2O3platelets instead of conventional Al2O3particles as a filler material was investigated. As the Al2O3platelets were arranged parallel to the casting direction during tape casting and had preferred orientation in the green tape, the linear shrinkage of the green body took place mainly in the direction perpendicular to the platelet planes with just a little shrinkage in the horizontal without any external force during sintering. The intended anisotropic shrinkage of the new LTCC system made the constrained sintering of 10‐mm‐thick ceramic body possible, which decreased the sintering shrinkage tolerance in thex–ydirections of the 10‐mm‐thick ceramic body from 0.5% to 0.05%. The new LTCC system showed equivalent electrical properties and remarkable physical properties in comparison with a conventional LTCC system.
Alumina Scale Formation: A New Perspective
Authors: Heuer Arthur H. , Hovis David B. , Smialek James L. , Gleeson Brian
Published: May 2011

Oxidation of Al2O3 scale‐forming alloys is of immense technological significance and has been a subject of much scientific inquiry for decades. The oxidation reaction is remarkably complex, involving issues of alloy composition, kinetics, thermodynamics, microstructure, mechanics and mechanical properties, crystallography, etc. A brief overview of the formation of passivating, thermally grown oxide Al2O3 scales will be given in the light of recent findings on the defect structure and associated transport behavior of α‐Al2O3. It is inferred that the electronic structure of Al2O3 is of direct relevance to understand the Al2O3 scale growth. We also discuss the effect of the so‐called “reactive” elements (REs)—Y, Zr, and Hf—on reducing the rate of Al2O3 scale thickening by reducing the outward flux of aluminum. An important aspect of the “new perspective” is the suggestion that the REs change the electronic structure of Al2O3—the relevant near‐band‐edge defect (grain boundary) states that are crucial to vacancy creation both at the scale/gas and scale/metal interfaces.