bluuebunny/crossref_metadata_2025 huggingface dataset:
Rapid Identification of the Mycobacterium tuberculosis Complex by Combining the ESAT-6/CFP-10 Immunochromatographic Assay and Smear Morphology
Authors: Shen Gwan-Han , Chiou Chien-Shun , Hu Shiau-Ting , Wu Kun-Ming , Chen Jiann-Hwa
Published: December 2010

Early secretory antigen 6 (ESAT-6) and cell filtrate protein 10 (CFP-10) are two antigens secreted as a complex by the replicating
Mycobacterium tuberculosis
complex (MTC). Recently, an immunochromatographic assay (ICA) using a monoclonal antibody against the ESAT-6/CFP-10 complex was developed for the purpose of MTC detection. In this study, the efficacy of the assay was tested with 603 BACTEC cultures that were incubated for 3 additional days after positive signals appeared in the BACTEC MGIT 960 system. Bacterial isolates were recovered from these 603 BACTEC cultures, and 332 MTC isolates, 270 nontuberculosis mycobacterial isolates, and 1
Nocardia
isolate were identified by using standard biochemical assays. The ESAT-6/CFP-10 assay detected 322 MTC cultures, resulting in a sensitivity of 97% and a specificity of 97.4%. To reduce the false-negative rate and improve the sensitivity, either serpentine cording in an acid-fast bacillus stain of the cultural smear, the ESAT-6/CFP-10 assay, or a combination of both was used for MTC detection. The sensitivity was then increased to 99.1%, and the negative predictive value increased to 98.9%, but the specificity decreased to 94.8% and the positive predictive value decreased to 95.9%. However, a combination of serpentine cording in cultural smears and the positivity of the ICA resulted in the specificity and positive predictive values of 100%. Therefore, BACTEC cultures with both serpentine cording and positivity of the ESAT-6/CFP-10 assay could be reported to contain MTC directly. The ESAT-6/CFP-10 assay may be an alternative of the Capilia assay (MPB64-ICA) as a convenient and cost-effective method for identification of MTC in culture.
Using Molecular Epidemiology To Trace Transmission of Nosocomial Norovirus Infection
Authors: Sukhrie Faizel H. A. , Beersma Matthias F. C. , Wong Albert , van der Veer Bas , Vennema Harry , Bogerman Jolanda , Koopmans Marion
Published: December 2010

Nosocomial norovirus (NoV) infection is common and may lead to complications in vulnerable hospitalized patients. Understanding sources and modes of transmission of noroviruses within health care settings will support the design of evidence-based strategies for reducing introduction and further spread. We sequenced a highly variable segment of the genome to identify possible clusters in patients with and without acute gastroenteritis who were hospitalized in the period 2002-2007. Admission and sampling dates were used to separate patients with nosocomial infection from those without nosocomial infection. Epidemiological clustering retrieved 22 clusters, defined as ≥2 patients with nosocomial infection on the same ward within 5 days. In total, 264 patients (of 2,458 tested) were diagnosed with NoV infection, and 61% of the patient strains could be genotyped. Of those, 51% (n= 82) belonged to GII.4, 34% (n= 54) belonged to GII.3, and 15% (n= 24) belonged to other genotypes (GI.6B, GII.17, GII.7, and GII.2). In children's wards, GII.3 strains were associated with nosocomial spread more often than other viruses were, whereas in adults this was the case for GII.4 strains. Sequence alignment recognized 11 new clusters based on identical P2 domains (4 GII.3 and 7 GII.4 clusters), involving patients in different wards. This increased the total number of recognized clusters by 50%. Five of these clusters involved at least one outpatient, providing a possible target for improvement of infection control. We concluded that the use of sequence-based typing should be considered for identifying hidden nosocomial clusters of NoV infections within health care settings.
Impact of Immigration on the Molecular Epidemiology of Tuberculosis in Rhode Island
Authors: Vanhomwegen Jessica , Kwara Awewura , Martin Melissa , Gillani Fizza S. , Fontanet Arnaud , Mutungi Peninnah , Crellin Joyce , Obaro Stephen , Gosciminski Michael , Carter E. Jane , Rastogi Nalin
Published: December 2010

While foreign-born persons constitute only 11% of the population in the state of Rhode Island, they account for more than 65% of incident tuberculosis (TB) annually. We investigated the molecular-epidemiological differences between foreign-born and U.S.-born TB patients to estimate the degree of recent transmission and identify predictors of clustering. A total of 288 isolates collected from culture-confirmed TB cases in Rhode Island between 1995 and 2004 were fingerprinted by spoligotyping and 12-locus mycobacterial interspersed repetitive units. Of the 288 fingerprinted isolates, 109 (37.8%) belonged to 36 genetic clusters. Our findings demonstrate that U.S.-born patients, Hispanics, Asian/Pacific islanders, and uninsured patients were significantly more likely to be clustered. Recent transmission among the foreign-born population was restricted and occurred mostly locally, within populations originating from the same region. Nevertheless, TB transmission between the foreign-born and U.S.-born population should not be neglected, since 80% of the mixed clusters of foreign- and U.S.-born persons arose from a foreign-born source case. We conclude that timely access to routine screening and treatment for latent TB infection for immigrants is vital for disease elimination in Rhode Island.
Unusual Case of Necrotizing Fasciitis Caused by Vibrio cholerae O137
Authors: Ottaviani Donatella , Leoni Francesca , Rocchegiani Elena , Canonico Cristina , Masini Laura , Pianetti Anna , Parlani Chiara , Luzzi Ida , Caola Iole , Paternoster Claudio , Carraturo Antonio
Published: December 2010

We report a case of necrotizing fasciitis caused by
Vibrio cholerae
O137 in an immunocompromised 49-year-old man. The infection was acquired following a minor traumatic injury and exposure to seawater during the summer of 2009 in Italy. Although highly immunocompromised, the patient survived. The strain was cytotoxic, invasive, and adhesive and contained a fragment of the El Tor-like hemolysin (El Tor
hlyA
) gene.
Interleukin-21-Producing HIV-1-Specific CD8 T Cells Are Preferentially Seen in Elite Controllers
Authors: Williams LaTonya D. , Bansal Anju , Sabbaj Steffanie , Heath Sonya L. , Song Wei , Tang Jianming , Zajac Allan J. , Goepfert Paul A.
Published: December 2010

A hallmark of human immunodeficiency virus type 1 (HIV-1) pathogenesis is the rapid loss of CD4 T cells leading to generalized immune dysfunction, including an exhausted CD8 T cell phenotype. Understanding the necessary factors that govern the functional quality and protective potential of antiviral T cell responses would facilitate rational vaccine design and improve therapeutic strategies to combat persistent infections. Mouse models of chronic viral infection demonstrate that interleukin-21 (IL-21), produced primarily by CD4 T cells, is required for the generation and maintenance of functionally competent CD8 T cells and viral containment. We reasoned that preserved IL-21 production during HIV-1 infection would be associated with enhanced CD8 T cell function, allowing improved viral control. Here we analyzed the ability of CD4 and CD8 T cells to produce several cytokines in addition to IL-21
ex vivo
following stimulation with overlapping HIV-1 peptides. Both CD4 and CD8 T cells were able to produce IL-21 in response to HIV-1 infection, with the latter cell type more closely associated with viral control. Furthermore, IL-21-producing HIV-1-specific CD4 T cells (compared to those producing other cytokines) were the best indicator of functional CD8 T cells. Our results demonstrate that HIV-1-specific IL-21-producing CD8 T cells are induced following primary infection and enriched in elite controllers, suggesting a critical role for these cells in the maintenance of viremia control.
Foamy Virus Nuclear RNA Export Is Distinct from That of Other Retroviruses
Authors: Bodem Jochen , Schied Tanja , Gabriel Richard , Rammling Matthias , Rethwilm Axel
Published: December 2010

Most retroviruses express all of their genes from a single primary transcript. In order to allow expression of more than one gene from this RNA, differential splicing is extensively used. Cellular quality control mechanisms retain and degrade unspliced or partially spliced RNAs in the nucleus. Two pathways have been described that explain how retroviruses circumvent this nuclear export inhibition. One involves a constitutive transport element in the viral RNA that interacts with the cellular mRNA transporter proteins NXF1 and NXT1 to facilitate nuclear export. The other pathway relies on the recognition of a viral RNA element by a virus-encoded protein that interacts with the karyopherin CRM1. In this report, we analyze the protein factors required for the nuclear export of unspliced foamy virus (FV) mRNA. We show that this export is CRM1 dependent. In contrast to other complex retroviruses, FVs do not encode an export-mediating protein. Cross-linking experiments indicated that the cellular protein HuR binds to the FV RNA. Inhibition studies showed that both ANP32A and ANP32B, which are known to bridge HuR and CRM1, are essential for FV RNA export. By using this export pathway, FVs solve a central problem of viral replication.
Identification of Two Functional Domains within the Arenavirus Nucleoprotein
Authors: Levingston Macleod Jesica M. , D'Antuono Alejandra , Loureiro Maria Eugenia , Casabona Juan Cruz , Gomez Guillermo A. , Lopez Nora
Published: December 2010

Tacaribe virus (TCRV) belongs to the
Arenaviridae
family. Its bisegmented negative-stranded RNA genome encodes the nucleoprotein (N), the precursor of the envelope glycoproteins, the polymerase (L), and a RING finger matrix (Z) protein. The 570-amino-acid N protein binds to viral RNA, forming nucleocapsids, which are the template for transcription and replication by the viral polymerase. We have previously shown that the interaction between N and Z is required for assembly of infectious virus-like particles (VLPs) (J. C. Casabona et al., J. Virol. 83:7029-7039, 2009). Here, we examine the functional organization of TCRV N protein. A series of deletions and point mutations were introduced into the N-coding sequence, and the ability of the mutants to sustain heterotypic (N-Z) or homotypic (N-N) interactions was analyzed. We found that N protein displays two functional domains. By using coimmunoprecipitation studies, VLP incorporation assays, and double immunofluorescence staining, the carboxy-terminal region of N was found to be required for N-Z interaction and also necessary for incorporation of N protein into VLPs. Moreover, further analysis of this region showed that the integrity of a putative zinc-finger motif, as well as its amino-flanking sequence (residues 461 to 489), are critical for Z binding and N incorporation into VLPs. In addition, we provide evidence of an essential role of the amino-terminal region of N protein for N-N interaction. In this regard, using reciprocal coimmunoprecipitation analysis, we identified a 28-residue region predicted to form a coiled-coil domain (residues 92 to 119) as a newly recognized molecular determinant of N homotypic interactions.
Phosphorylation and Polyubiquitination of Transforming Growth Factor β-Activated Kinase 1 Are Necessary for Activation of NF-κB by the Kaposi's Sarcoma-Associated Herpesvirus G Protein-Coupled Receptor
Authors: Bottero Virginie , Kerur Nagaraj , Sadagopan Sathish , Patel Kinjan , Sharma-Walia Neelam , Chandran Bala
Published: December 2010

Kaposi's sarcoma-associated herpesvirus (KSHV) G protein-coupled receptor (vGPCR) protein has been shown to induce several signaling pathways leading to the modulation of host gene expression. The hijacking of these pathways facilitates the viral life cycle and leads to tumorigenesis. In the present work, we show that transforming growth factor β (TGF-β)-activated kinase 1 (TAK1) is an important player in NF-κB activation induced by vGPCR. We observed that the expression of an inactive TAK1 kinase mutant (TAK1M) reduces vGPCR-induced NF-κB nuclear translocation and transcriptional activity. Consequently, the expression of several NF-κB target genes normally induced by vGPCR was blocked by TAK1M expression, including interleukin 8 (IL-8), Gro1, IκBα, COX-2, cIAP2, and Bcl2 genes. Similar results were obtained after downregulation of TAK1 by small interfering RNA (siRNA) technology. The expression of vGPCR recruited TAK1 to the plasma membrane, and vGPCR interacts with TAK1. vGPCR expression also induced TAK1 phosphorylation and lysine 63-linked polyubiquitination, the two markers of the kinase's activation. Finally, inhibition of TAK1 by celastrol inhibited vGPCR-induced NF-κB activation, indicating this natural compound could be used as a potential therapeutic drug against KSHV malignancies involving vGPCR.
Cell-Associated Viral Burden Provides Evidence of Ongoing Viral Replication in Aviremic HIV-2-Infected Patients
Authors: Soares Rui S. , Tendeiro Rita , Foxall Russell B. , Baptista António P. , Cavaleiro Rita , Gomes Perpétua , Camacho Ricardo , Valadas Emília , Doroana Manuela , Lucas Margarida , Antunes Francisco , Victorino Rui M. M. , Sousa Ana E.
Published: December 2010

Viremia is significantly lower in HIV-2 than in HIV-1 infection, irrespective of disease stage. Nevertheless, the comparable proviral DNA burdens observed for these two infections indicate similar numbers of infected cells. Here we investigated this apparent paradox by assessing cell-associated viral replication. We found that untreated HIV-1-positive (HIV-1+) and HIV-2+individuals, matched for CD4 T cell depletion, exhibited similargagmRNA levels, indicating that significant viral transcription is occurring in untreated HIV-2+patients, despite the reduced viremia (undetectable to 2.6 × 104RNA copies/ml). However,tatmRNA transcripts were observed at significantly lower levels in HIV-2+patients, suggesting that the rate ofde novoinfection is decreased in these patients. Our data also reveal a direct relationship ofgagandtattranscripts with CD4 and CD8 T cell activation, respectively. Antiretroviral therapy (ART)-treated HIV-2+patients showed persistent viral replication, irrespective of plasma viremia, possibly contributing to the emergence of drug resistance mutations, persistent hyperimmune activation, and poor CD4 T cell recovery that we observed with these individuals. In conclusion, we provide here evidence of significant ongoing viral replication in HIV-2+patients, further emphasizing the dichotomy between amount of plasma virus and cell-associated viral burden and stressing the need for antiretroviral trials and the definition of therapeutic guidelines for HIV-2 infection.
Phylogeography of West Nile Virus: from the Cradle of Evolution in Africa to Eurasia, Australia, and the Americas
Authors: May Fiona J. , Davis C. Todd , Tesh Robert B. , Barrett Alan D. T.
Published: December 2010

West Nile virus (WNV) is the most widely distributed of the encephalitic flaviviruses and is a major cause of encephalitis, with isolates obtained from all continents, apart from Antarctica. Subsequent to its divergence from the other members of the Japanese encephalitis virus complex, presumably in Africa, WNV has diverged into individual lineages that mostly correspond with geographic distribution. Here we elucidate the phylogeography and evolutionary history of isolates from lineage 1 of WNV. Interestingly, there are many examples of the same amino acid having evolved independently on multiple occasions. In Africa, WNV exists in an endemic cycle, whereas it is epidemic in Europe, being reintroduced regularly from Africa either directly (in western Europe) or via the Middle East (in eastern Europe). Significantly, introduction into other geographic areas has occurred on one occasion only in each region, leading to subsequent establishment and expansion of the virus in these areas. Only one endemic genotype each is present in India and Australia, suggesting that WNV was successfully introduced into these locations once only. Each introduction occurred many centuries ago, probably due to trade and exploration during the 19th century. Likewise, in the Americas, WNV was successfully introduced in 1999 and subsequently became endemic across most temperate regions of North America (NA). In contrast to previous suggestions, an isolate from the epidemic in Israel in 1998 was not the direct progenitor of the NA epidemic; rather, both epidemics originated from the same (unknown) location.
Functional Characterization of Kaposi's Sarcoma-Associated Herpesvirus Open Reading Frame K8 by Bacterial Artificial Chromosome-Based Mutagenesis
Authors: Wang Yan , Sathish Narayanan , Hollow Charles , Yuan Yan
Published: December 2010

The open reading frame K8 of Kaposi's sarcoma-associated herpesvirus (KSHV) encodes a basic leucine zipper (bZip) protein that binds to the origin of viral DNA replication and is an integral component of viral lytic DNA replication complex. Moreover, K8 physically interacts with replication and transcription activator (RTA) and represses its transactivation activity on several viral promoters. To investigate the role of this protein in viral life cycle, we constructed two K8-null recombinant mutant viruses (BAC-ΔK8 and BAC-stopK8) by using a bacterial artificial chromosome (BAC) system. Latent viral infection can be reconstituted in 293T and BJAB cells with wild-type and the K8-null recombinant viruses by introducing the cloned viral genomes into the cells. When the cells carrying these viruses were induced with 12-
O
-tetradecanoylphorbol-13-acetate (TPA) and sodium butyrate, no significant difference was seen in overall viral gene expression between wild-type and K8-null viruses, with lytic DNA replication still active in the latter. However, 293T cells harboring K8-null mutant viruses, either BAC-ΔK8 or BAC-stopK8, displayed lower copy numbers of latent KSHV genome in comparison with wild-type viruses. Furthermore, although K8 deficiency appeared to not affect infectivity when K8-null viruses were used to infect 293T, primary human microvascular dermal endothelial and human foreskin fibroblast cells, they exhibited much lower viral genome copy numbers in all types of cell compared to wild-type viruses. Taken together, these data suggest a possible role of K8 in abortive lytic DNA replication occurring in early stages of
de novo
infection or in the maintenance of latent viral genomes.
Fibronectin upregulates cGMP-dependent protein kinase type Iβ through C/EBP transcription factor activation in contractile cells
Authors: Chamorro-Jorganes Aranzazu , Calleros Laura , Griera Mercedes , Saura Marta , Luengo Alicia , Rodriguez-Puyol D. , Rodriguez-Puyol M.
Published: December 2010

The nitric oxide (NO)-soluble guanylate cyclase (sGC) pathway exerts most of its cellular actions through the activation of the cGMP-dependent protein kinase (PKG). Accumulation of extracellular matrix is one of the main structural changes in pathological conditions characterized by a decreased activity of this pathway, such as hypertension, diabetes, or aging, and it is a well-known fact that extracellular matrix proteins modulate cell phenotype through the interaction with membrane receptors such as integrins. The objectives of this study were 1) to evaluate whether extracellular matrix proteins, particularly fibronectin (FN), modulate PKG expression in contractile cells, 2) to analyze the mechanisms involved, and 3) to evaluate the functional consequences. FN increased type I PKG (PKG-I) protein content in human mesangial cells, an effect dependent on the interaction with β1-integrin. The FN upregulation of PKG-I protein content was due to increased mRNA expression, determined by augmented transcriptional activity of the PKG-I promoter region. Akt and the transcription factor CCAAT enhancer-binding protein (C/EBP) mediated the genesis of these changes. FN also increased PKG-I in another type of contractile cell, rat vascular smooth muscle cells (RVSMC). Tirofiban, a pharmacological analog of FN, increased PKG-I protein content in RVSMC and rat aortic walls and magnified the hypotensive effect of dibutyryl cGMP in conscious Wistar rats. The present results provide evidence of a mechanism able to increase PKG-I protein content in contractile cells. Elucidation of this novel mechanism provides a rationale for future pharmacotherapy in certain vascular diseases.
Fragmented mitochondria are sensitized to Bax insertion and activation during apoptosis
Authors: Brooks Craig , Cho Sung-Gyu , Wang Cong-Yi , Yang Tianxin , Dong Zheng
Published: December 2010

Recent studies have shown mitochondrial fragmentation during cell stress and have suggested a role for the morphological change in mitochondrial injury and ensuing apoptosis. However, the underlying mechanism remains elusive. Here we demonstrate that mitochondrial fragmentation facilitates Bax insertion and activation in mitochondria, resulting in the release of apoptogenic factors. In HeLa cells, overexpression of mitofusins attenuated mitochondrial fragmentation during cisplatin- and azide-induced cell injury, which was accompanied by less apoptosis and less cytochrome c release from mitochondria. Similar effects were shown by inhibiting the mitochondrial fission protein Drp1 with a dominant negative mutant (dn-Drp1). Mitofusins and dn-Drp1 did not seem to significantly affect Bax translocation/accumulation to mitochondria; however, they blocked Bax insertion and activation in mitochondrial membrane. Consistently, in rat kidney proximal tubular cells, small interfering RNA knockdown of Drp1 prevented mitochondrial fragmentation during azide-induced ATP depletion, which was accompanied by less Bax activation, insertion, and oligomerization in mitochondria. These cells released less cytochrome c and AIF from mitochondria and showed significantly lower apoptosis. Finally, mitofusin-null mouse embryonic fibroblasts (MEF) had fragmented mitochondria. These MEFs were more sensitive to cisplatin-induced Bax activation, release of cytochrome c, and apoptosis. Together, this study provides further support for a role of mitochondrial fragmentation in mitochondrial injury and apoptosis. Mechanistically, mitochondrial fragmentation may sensitize the cells to Bax insertion and activation in mitochondria, facilitating the release of apoptogenic factors and consequent apoptosis.
Loss of Rab27 function results in abnormal lung epithelium structure in mice
Authors: Bolasco Giulia , Tracey-White Dhani C. , Tolmachova Tanya , Thorley Andrew J. , Tetley Teresa D. , Seabra Miguel C. , Hume Alistair N.
Published: December 2010

Rab27 small GTPases regulate secretion and movement of lysosome-related organelles such as T cell cytolytic granules and platelet-dense granules. Previous studies indicated that Rab27a and Rab27b are expressed in the murine lung suggesting that they regulate secretory processes in the lung. Consistent with those studies, we found that Rab27a and Rab27b are expressed in cell types that contain secretory granules: alveolar epithelial type II (AEII) and Clara cells. We then used Rab27a/Rab27b double knockout (DKO) mice to examine the functional consequence of loss of Rab27 proteins in the murine lung. Light and electron microscopy revealed a number of morphological changes in lungs from DKO mice when compared with those in control animals. In aged DKO mice we observed atrophy of the bronchiolar and alveolar epithelium with reduction of cells numbers, thinning of the bronchiolar epithelium and alveolar walls, and enlargement of alveolar airspaces. In these samples we also observed increased numbers of activated foamy alveolar macrophages and granulocyte containing infiltrates together with reduction in the numbers of Clara cells and AEII cells compared with control. At the ultrastructural level we observed accumulation of cytoplasmic membranes and vesicles in Clara cells. Meanwhile, AEII cells in DKO accumulated large mature lamellar bodies and lacked immature/precursor lamellar bodies. We hypothesize that the morphological changes observed at the ultrastructural level in DKO samples result from secretory defects in AEII and Clara cells and that over time these defects lead to atrophy of the epithelium.
Muscle fiber recruitment and the slow component of O2uptake: constant work rate vs. all-out sprint exercise
Authors: Vanhatalo Anni , Poole David C. , DiMenna Fred J. , Bailey Stephen J. , Jones Andrew M.
Published: December 2010

The slow component of pulmonary O2uptake (V̇o2) during constant work rate (CWR) high-intensity exercise has been attributed to the progressive recruitment of (type II) muscle fibers. We tested the following hypotheses: 1) the V̇o2slow component gain would be greater in a 3-min all-out cycle test than in a work-matched CWR test, and 2) the all-out test would be associated with a progressive decline, and the CWR test with a progressive increase, in muscle activation, as estimated from the electromyogram (EMG) of the vastus lateralis muscle. Eight men (aged 21–39 yr) completed a ramp incremental test, a 3-min all-out test, and a work- and time-matched CWR test to exhaustion. The maximum V̇o2attained in an initial ramp incremental test (3.97 ± 0.83 l/min) was reached in both experimental tests (3.99 ± 0.84 and 4.03 ± 0.76 l/min for all-out and CWR, respectively). The V̇o2slow component was greater ( P < 0.05) in the all-out test (1.21 ± 0.31 l/min, 4.2 ± 2.2 ml·min−1·W−1) than in the CWR test (0.59 ± 0.22 l/min, 1.70 ± 0.5 ml·min−1·W−1). The integrated EMG declined by 26% ( P < 0.001) during the all-out test and increased by 60% ( P < 0.05) during the CWR test from the first 30 s to the last 30 s of exercise. The considerable reduction in muscle efficiency in the all-out test in the face of a progressively falling integrated EMG indicates that progressive fiber recruitment is not requisite for development of the V̇o2slow component during voluntary exercise in humans.
M1 and M4 Receptors Modulate Hippocampal Pyramidal Neurons
Authors: Dasari Sameera , Gulledge Allan T.
Published: December 2010

Acetylcholine (ACh), acting at muscarinic ACh receptors (mAChRs), modulates the excitability and synaptic connectivity of hippocampal pyramidal neurons. CA1 pyramidal neurons respond to transient (“phasic”) mAChR activation with biphasic responses in which inhibition is followed by excitation, whereas prolonged (“tonic”) mAChR activation increases CA1 neuron excitability. Both phasic and tonic mAChR activation excites pyramidal neurons in the CA3 region, yet ACh suppresses glutamate release at the CA3-to-CA1 synapse (the Schaffer–collateral pathway). Using mice genetically lacking specific mAChRs (mAChR knockout mice), we identified the mAChR subtypes responsible for cholinergic modulation of hippocampal pyramidal neuron excitability and synaptic transmission. Knockout of M1 receptors significantly reduced, or eliminated, most phasic and tonic cholinergic responses in CA1 and CA3 pyramidal neurons. On the other hand, in the absence of other Gq-linked mAChRs (M3 and M5), M1 receptors proved sufficient for all postsynaptic cholinergic effects on CA1 and CA3 pyramidal neuron excitability. M3 receptors were able to participate in tonic depolarization of CA1 neurons, but otherwise contributed little to cholinergic responses. At the Schaffer–collateral synapse, bath application of the cholinergic agonist carbachol suppressed stratum radiatum–evoked excitatory postsynaptic potentials (EPSPs) in wild-type CA1 neurons and in CA1 neurons from mice lacking M1 or M2 receptors. However, Schaffer–collateral EPSPs were not significantly suppressed by carbachol in neurons lacking M4 receptors. We therefore conclude that M1 and M4 receptors are the major mAChR subtypes responsible for direct cholinergic modulation of the excitatory hippocampal circuit.
Kv3-Like Potassium Channels Are Required for Sustained High-Frequency Firing in Basal Ganglia Output Neurons
Authors: Ding Shengyuan , Matta Shannon G. , Zhou Fu-Ming
Published: December 2010

The GABA projection neurons in the substantial nigra pars reticulata (SNr) are key output neurons of the basal ganglia motor control circuit. These neurons fire sustained high-frequency, short-duration spikes that provide a tonic inhibition to their targets and are critical to movement control. We hypothesized that a robust voltage-activated K+conductance that activates quickly and resists inactivation is essential to the remarkable fast-spiking capability in these neurons. Semi-quantitative RT-PCR (qRT-PCR) analysis on laser capture-microdissected nigral neurons indicated that mRNAs for Kv3.1 and Kv3.4, two key subunits for forming high activation threshold, fast-activating, slow-inactivating, 1 mM tetraethylammonium (TEA)-sensitive, fast delayed rectifier ( IDR-fast) type Kv channels, are more abundant in fast-spiking SNr GABA neurons than in slow-spiking nigral dopamine neurons. Nucleated patch clamp recordings showed that SNr GABA neurons have a strong Kv3-like IDR-fastcurrent sensitive to 1 mM TEA that activates quickly at depolarized membrane potentials and is resistant to inactivation. IDR-fastis smaller in nigral dopamine neurons. Pharmacological blockade of IDR-fastby 1 mM TEA impaired the high-frequency firing capability in SNr GABA neurons. Taken together, these results indicate that Kv3-like channels mediating fast-activating, inactivation-resistant IDR-fastcurrent are critical to the sustained high-frequency firing in SNr GABA projection neurons and hence movement control.
DNA Vaccination: Using the Patient′s Immune System to Overcome Cancer
Authors: Eschenburg Georg , Stermann Alexander , Preissner Robert , Meyer Hellmuth-Alexander , Lode Holger N.
Published: December 2010

Cancer is one of the most challenging diseases of today. Optimization of standard treatment protocols consisting of the main columns of chemo‐ and radiotherapy followed or preceded by surgical intervention is often limited by toxic side effects and induction of concomitant malignancies and/or development of resistant mechanisms. This requires the development of therapeutic strategies which are as effective as standard therapies but permit the patients a life without severe negative side effects. Along this line, the development of immunotherapy in general and the innovative concept of DNA vaccination in particular may provide a venue to achieve this goal. Using the patient′s own immune system by activation of humoral and cellular immune responses to target the cancer cells has shown first promising results in clinical trials and may allow reduced toxicity standard therapy regimen in the future. The main challenge of this concept is to transfer the plethora of convincing preclinical and early clinical results to an effective treatment of patients.
Calculation of the Rydberg Energy Levels for Francium Atom
Authors: Shizhong Huang , Qiufeng Sun
Published: December 2010

Based on the weakest bound electron potential model theory, the Rydberg energy levels and quantum defects of nsS21/2(n=8--50), ndD23/2(n=6--50), and ndD25/2(n=6--50) spectrum series for francium atom are calculated. The calculated results are in excellent agreement with the 74 known experimentally measured levels (the absolute difference is less than 0.03 cm-1) and 58 energy levels for highly excited states are predicted.
Increased Daytime Sleepiness in Patients with Childhood Craniopharyngioma and Hypothalamic Tumor Involvement: Review of the Literature and Perspectives
Authors: Müller Hermann L.
Published: December 2010

Childhood craniopharyngiomas are rare embryogenic malformations of the sellar region, presumably derived from Rathke cleft epithelium. The overall survival rates after neurosurgical intervention and/or irradiation are high (92%). However, the quality of survival is frequently impaired due to endocrine deficiencies, sleep disturbances, daytime sleepiness, and severe obesity caused by hypothalamic lesions. Based on self-assessment using nutritional diaries, caloric intake was similar in patients and BMI-matched controls. Analyses of physical activity by accelerometric measurements showed a markedly lower level of physical activity. Significant daytime sleepiness and disturbances of circadian rhythms have been demonstrated in obese childhood craniopharyngioma patients. Daytime sleepiness and obesity in these patients were both correlated with low nocturnal and early morning melatonin levels. Polysomnographic studies in patients with severe daytime sleepiness revealed sleeping patterns typical for secondary narcolepsy. Reports on a beneficial effect of treatment with central stimulating agents supported the hypothesis that secondary narcolepsy should be considered as a rare cause for severe daytime sleepiness in patients with childhood craniopharyngioma.
Inhibition of RNA Helicases of ssRNA+Virus Belonging to Flaviviridae, Coronaviridae and Picornaviridae Families
Authors: Briguglio Irene , Piras Sandra , Corona Paola , Carta Antonio
Published: December 2010

Many viral pathogens encode the motor proteins named RNA helicases which display various functions in genome replication. General strategies to design specific and selective drugs targeting helicase for the treatment of viral infections could actviaone or more of the following mechanisms: inhibition of the NTPase activity, by interferences with ATP binding and therefore by limiting the energy required for the unwinding and translocation, or by allosteric mechanism and therefore by stabilizing the conformation of the enzyme in low helicase activity state; inhibition of nucleic acids binding to the helicase; inhibition of coupling of ATP hydrolysis to unwinding; inhibition of unwinding by sterically blocking helicase translocation. Recently, byin vitroscreening studies, it has been reported that several benzotriazole, imidazole, imidazodiazepine, phenothiazine, quinoline, anthracycline, triphenylmethane, tropolone, pyrrole, acridone, small peptide, and Bananin derivatives are endowed with helicase inhibition of pathogen viruses belonging to Flaviviridae, Coronaviridae, and Picornaviridae families.
Controlled On-Chip Single-Photon Transfer Using Photonic Crystal Coupled-Cavity Waveguides
Authors: Seigneur Hubert Pascal , Weed Matthew , Leuenberger Michael Niklaus , Schoenfeld Winston Vaughan
Published: December 2010

To the end of realizing a quantum network on-chip, single photons must be guided consistently to their proper destination both on demand and without alteration to the information they carry. Coupled cavity waveguides are anticipated to play a significant role in this regard for two important reasons. First, these structures can easily be included within fully quantum-mechanical models using the phenomenological description of the tight-binding Hamiltonian, which is simply written down in the basis of creation and annihilation operators that move photons from one quasimode to another. This allows for a deeper understanding of the underlying physics and the identification and characterization of features that are truly critical to the behavior of the quantum network using only a few parameters. Second, their unique dispersive properties together with the careful engineering of the dynamic coupling between nearest neighbor cavities provide the necessary control for high-efficiency single-photon on-chip transfer. In this publication, we report transfer efficiencies in the upwards of 93% with respect to a fully quantum-mechanical approach and unprecedented 77% in terms of transferring the energy density contained in a classical quasibound mode from one cavity to another.
Development of a Serum Biomarker Assay That Differentiates Tumor‐Associated MUC5AC (NPC‐1C ANTIGEN) from Normal MUC5AC
Authors: Luka Janos , Arlen Philip M. , Bristol Andrew
Published: December 2010

A serum ELISA using a monoclonal antibody that detects a MUC5AC‐related antigen (NPC‐1C antigen) expressed by pancreatic and colorectal cancer was developed. The NPC‐1C antibody reacts with specific epitopes expressed by tumor‐associated MUC5AC that does not appear on MUC5AC from normal tissues. Based on observations of a highly specific antibody, we tested the ELISA to differentiate serum from healthy blood donors compared to serum from patients with colorectal or pancreatic cancer. Additionally, patient tumor tissue was stained to examine the expression pattern of MUC5AC‐related antigen in pancreatic and colorectal cancers. The results indicate the NPC‐1C antibody ELISA distinguished serum of cancer patients from normal donors with very good sensitivity and specificity. Most patient′s tumor biopsy exhibited NPC‐1C antibody reactivity, indicating that tumor‐associated MUC5AC antigen from tumor is shed into blood, where it can be detected by the NPC‐1C antibody ELISA. This serum test provides a new tool to aid in the diagnosis of these cancers and immune monitoring of cancer treatment regimens.
The Best Estimate Plus Uncertainty Challenge in the Current Licensing Process of Present Reactors
Authors: D′Auria Francesco , Mazzantini Oscar
Published: December 2010

Within the licensing process of the KWU Atucha II PHWR (Pressurized Heavy Water Reactor), the BEPU (Best Estimate Plus Uncertainty) approach has been selected for issuing of the Chapter 15 on FSAR (Final Safety Analysis Report). The key steps of the entire process are basically two: (a) the selection of PIE (Postulated Initiating Events) and (b) the analysis by best estimate models supported by uncertainty evaluation. Otherwise, key elements of the approach are (1) availability of qualified computational tools including suitable uncertainty method, (2) demonstration of quality, and (3) acceptability and endorsement by the licensing authority. The effort of issuing Chapter 15 is terminated at the time of issuing of the present paper, and the safety margins available for the operation of the concerned NPP (Nuclear Power Plant) have been quantified.
The Cyclical Integration Model as a Way of Managing Major Educational Change
Authors: Berlach Richard G.
Published: December 2010

Where minds meet, there lies the change vector. With regard to change management, however, minds regularly fail to meet in the crucial change space. They either unwittingly zip past each other, deliberately avoid one another, or worse still, collide with excruciating impact. This paper examines the interrelated role of government, the public service and professionals in successfully transitioning major change initiatives. It is argued that unless these agencies operate in synchrony, change negotiation is likely to be hampered. To this end, a model of “Cyclical Integration” is presented and supported by driving questions for each of the three agencies facilitating the change process. Although it is considered that these questions are sufficiently broad to encompass change management across any number of enterprises, what is being considered in this paper is the arena of compulsory education.
Molecular Mechanism of MART-1+/A*0201+ Human Melanoma Resistance to Specific CTL-Killing Despite Functional Tumor–CTL Interaction
Authors: Jazirehi Ali R. , Baritaki Stavroula , Koya Richard C. , Bonavida Benjamin , Economou James S.
Published: December 2010

Durable responses in metastatic melanoma patients remain generally difficult to achieve. Adoptive cell therapy (ACT) with ex vivo engineered lymphocytes expressing high affinity T-cell receptors (TCRα/β) for the melanoma antigen MART-127–35/HLA-A*0201 [recognized by F5 cytotoxic T lymphocytes (F5 CTL)] has been found to benefit certain patients. However, many other patients are inherently unresponsive and/or relapse for unknown reasons. To analyze the basis for the acquired resistance and strategies to reverse it, we established F5 CTL–resistant (R) human melanoma clones from relatively sensitive parental lines under selective F5 CTL pressure. Surface MART-127–35/HLA-A*0201 in these clones was unaltered and F5 CTLs recognized and interacted with them similar to the parental lines. Nevertheless, the R clones were resistant to F5 CTL killing, exhibited hyperactivation of the NF-κB survival pathway, and overexpression of the antiapoptotic genes B cell lymphoma protein 2 (Bcl-2), Bcl-2 related gene (long alternatively spliced variant of Bcl-x gene; Bcl-xL), and myeloid cell differentiation 1 (Mcl-1). Sensitivity to F5 CTL-killing could be increased by pharmacological inhibition of the NF-κB pathway, Bcl-2 family members, or the proteasome, the latter of which reduced NF-κB activity and diminished antiapoptotic gene expression. Specific gene-silencing (by siRNA) confirmed the protective role of antiapoptotic factors by reversing R clone resistance. Together, our findings suggest that long-term immunotherapy may impose a selection for the development of resistant cells that are unresponsive to highly avid and specific melanoma-reactive CTLs, despite maintaining expression of functional peptide:MHC complexes, due to activation of antiapoptotic signaling pathways. Though unresponsive to CTL, our results argue that resistant cells can be resensitized to immunotherapy with coadministration of targeted inhibitors to antiapoptotic survival pathways. Cancer Res; 71(4); 1406–17. ©2010 AACR.
Targeting RNA Polymerase I with an Oral Small Molecule CX-5461 Inhibits Ribosomal RNA Synthesis and Solid Tumor Growth
Authors: Drygin Denis , Lin Amy , Bliesath Josh , Ho Caroline B. , O'Brien Sean E. , Proffitt Chris , Omori Mayuko , Haddach Mustapha , Schwaebe Michael K. , Siddiqui-Jain Adam , Streiner Nicole , Quin Jaclyn E. , Sanij Elaine , Bywater Megan J. , Hannan Ross D. , Ryckman David , Anderes Kenna , Rice William G.
Published: December 2010

Deregulated ribosomal RNA synthesis is associated with uncontrolled cancer cell proliferation. RNA polymerase (Pol) I, the multiprotein complex that synthesizes rRNA, is activated widely in cancer. Thus, selective inhibitors of Pol I may offer a general therapeutic strategy to block cancer cell proliferation. Coupling medicinal chemistry efforts to tandem cell- and molecular-based screening led to the design of CX-5461, a potent small-molecule inhibitor of rRNA synthesis in cancer cells. CX-5461 selectively inhibits Pol I–driven transcription relative to Pol II–driven transcription, DNA replication, and protein translation. Molecular studies demonstrate that CX-5461 inhibits the initiation stage of rRNA synthesis and induces both senescence and autophagy, but not apoptosis, through a p53-independent process in solid tumor cell lines. CX-5461 is orally bioavailable and demonstrates in vivo antitumor activity against human solid tumors in murine xenograft models. Our findings position CX-5461 for investigational clinical trials as a potent, selective, and orally administered agent for cancer treatment. Cancer Res; 71(4); 1418–30. ©2010 AACR.
B7-H1 Overexpression Regulates Epithelial–Mesenchymal Transition and Accelerates Carcinogenesis in Skin
Authors: Cao Yujia , Zhang Lu , Kamimura Yosuke , Ritprajak Patcharee , Hashiguchi Masaaki , Hirose Sachiko , Azuma Miyuki
Published: December 2010

B7-H1 (CD274) is a T-cell coinhibitory molecule that is also often induced on human carcinoma cells, where its expression has been implicated in immune escape. Under inflammatory conditions, B7-H1 is also inducible in normal epithelial cells but little is known about its involvement in conversion of normal cells to tumor cells. Here, we show that skin-specific expression of B7-H1 accelerates inflammatory carcinogenesis in a methylcholantrene (MCA)-induced model of squamous cell carcinoma (SCC). Inflammatory responses induced by MCA or phorbol ester TPA were clearly inhibited in B7-H1 transgenic mice (B7-H1tg mice). Antibody-mediated blockade of either B7-H1 or the related molecule PD-1 revealed that their ability to limit inflammation relied on ligand interactions made by B7-H1 or PD-1. Skin keratinocytes derived from B7-H1tg mice exhibited constitutive reduction of E-cadherin, and SCC induced in B7-H1tg mice also showed loss of E-cadherin along with elevated expression of the transcription factors Slug and Twist that drive epithelial–mesenchymal transition (EMT). Our results indicate that upregulation of B7-H1 in skin epithelial cells promotes EMT and accelerates carcinogenesis, revealing insights into the significance of B7-H1 overexpression on solid tumor cells and hinting at a close relationship between EMT and immune escape signaling pathways in cancer. Cancer Res; 71(4); 1235–43. ©2010 AACR.
Induction of Monocyte Chemoattractant Protein-1 and Interleukin-10 by TGFβ1 in Melanoma Enhances Tumor Infiltration and Immunosuppression
Authors: Díaz-Valdés Nancy , Basagoiti María , Dotor Javier , Aranda Fernando , Monreal Iñaki , Riezu-Boj José Ignacio , Borrás-Cuesta Francisco , Sarobe Pablo , Feijoó Esperanza
Published: December 2010

Melanoma progression is associated with the expression of different growth factors, cytokines, and chemokines. Because TGFβ1 is a pleiotropic cytokine involved not only in physiologic processes but also in cancer development, we analyzed in A375 human melanoma cells, the effect of TGFβ1 on monocyte chemoattractant protein-1 (MCP-1) and interleukin-10 (IL-10) expression, two known factors responsible for melanoma progression. TGFβ1 increased the expression of MCP-1 and IL-10 in A375 cells, an effect mediated by the cross-talk between Smad, PI3K (phosphoinositide 3-kinase)/AKT, and BRAF-MAPK (mitogen activated protein kinase) signaling pathways. Supernatants from TGFβ1-treated A375 cells enhanced MCP-1–dependent migration of monocytes, which, in turn, expressed high levels of TGF,β1, bFGF, and VEGF mRNA. Moreover, these supernatants also inhibited functional properties of dendritic cells through IL-10–dependent mechanisms. When using in vitro, the TGFβ1-blocking peptide P144, TGFβ1-dependent Smad3 phosphorylation, and expression of MCP-1 and IL-10 were inhibited. In vivo, treatment of A375 tumor–bearing athymic mice with P144 significantly reduced tumor growth, associated with a lower macrophage infiltrate and decreased intratumor MCP-1 and VEGF levels, as well as angiogenesis. Finally, in C57BL/6 mice with B16-OVA melanoma tumors, when administered with immunotherapy, P144 decreased tumor growth and intratumor IL-10 levels, linked to enhanced activation of dendritic cells and natural killer cells, as well as anti-OVA T-cell responses. These results show new effects of TGFβ1 on melanoma cells, which promote tumor progression and immunosuppression, strongly reinforcing the relevance of this cytokine as a molecular target in melanoma. Cancer Res; 71(3); 812–21. ©2011 AACR.
Fes Tyrosine Kinase Expression in the Tumor Niche Correlates with Enhanced Tumor Growth, Angiogenesis, Circulating Tumor Cells, Metastasis, and Infiltrating Macrophages
Authors: Zhang Shengnan , Chitu Violeta , Stanley E. Richard , Elliott Bruce E. , Greer Peter A.
Published: December 2010

Fes is a protein tyrosine kinase with cell autonomous oncogenic activities that are well established in cell culture and animal models, but its involvement in human cancer has been unclear. Abundant expression of Fes in vascular endothelial cells and myeloid cell lineages prompted us to explore roles for Fes in the tumor microenvironment. In an orthotopic mouse model of breast cancer, we found that loss of Fes in the host correlated with reductions in engrafted tumor growth rates, metastasis, and circulating tumor cells. The tumor microenvironment in Fes-deficient mice also showed reduced vascularity and fewer macrophages. In co-culture with tumor cells, Fes-deficient macrophages also poorly promoted tumor cell invasive behavior. Taken together, our observations argue that Fes inhibition might provide therapeutic benefits in breast cancer, in part by attenuating tumor-associated angiogenesis and the metastasis-promoting functions of tumor-associated macrophages. Cancer Res; 71(4); 1465–73. ©2010 AACR.