bluuebunny/crossref_metadata_2025 huggingface dataset:
Revisiting Pneumococcal Carriage by Use of Broth Enrichment and PCR Techniques for Enhanced Detection of Carriage and Serotypes
Authors: Carvalho Maria da Gloria , Pimenta Fabiana C. , Jackson Delois , Roundtree Alexis , Ahmad Yusra , Millar Eugene V. , O'Brien Katherine L. , Whitney Cynthia G. , Cohen Adam L. , Beall Bernard W.
Published: March 2010

The measurement of pneumococcal carriage in the nasopharyngeal reservoir is subject to potential confounders that include low-density and multiple-strain colonization. To compare different methodologies, we picked a random sampling of 100 nasopharyngeal specimens recovered from infants less than 2 years of age who were previously assessed for pneumococcal carriage and serotypes by a conventional method that used direct plating from the transport/storage medium (50 specimens were culture negative and 50 specimens were culture positive for pneumococci). We used a broth enrichment approach and a conventional PCR approach (with and without broth enrichment) to determine pneumococcal carriage and serotypes, and the results were compared to the initial conventional culture-based results. Additionally, we used a
lytA
-targeted real-time PCR for pneumococcal detection. Broth enrichment for both the culture-based and the PCR-based methods enhanced the isolation of pneumococci and detection of serotype diversity, with the most effective serotype deduction method being one that used broth enrichment prior to sequential multiplex PCR. Similarly, we also found that broth enrichment followed by the
lytA
-specific real-time PCR was the most sensitive for the detection of apparent pneumococcal carriage. The broth enrichment, conventional multiplex PCR, and real-time PCR approaches used in this study were effective in detecting pneumococcal carriage in the 50 specimens that were negative by conventional direct plating from transport medium (range of numbers of positive specimens, 8/50 to 22/50 [16 to 44%]), and the three different serotyping approaches that used broth enrichment increased the number of serotype identifications from the 100 specimens (12 to 29 additional serotype identifications to be positive). A PCR-based approach that employed a broth enrichment step appeared to best enhance the detection of mixed serotypes and low-density pneumococcal carriage.
Rapid Detection of Influenza A Pandemic (H1N1) 2009 Virus Neuraminidase Resistance Mutation H275Y by Real-Time Reverse Transcriptase PCR
Authors: Hindiyeh Musa , Ram Daniela , Mandelboim Michal , Meningher Tal , Hirsh Shira , Robinov Jana , Levy Virginia , Orzitzer Sara , Azar Roberto , Grossman Zehava , Mendelson Ella
Published: March 2010

The emergence of oseltamivir-resistant influenza A pandemic (H1N1) 2009 virus highlights the need for rapid oseltamivir resistance screening. We report the development and validation of high-throughput real-time reverse transcriptase PCR assays for the detection of the H275Y substitution in the neuraminidase 1 gene that can be accomplished in 3 to 4 h.
The Human Cytomegalovirus UL36 Gene Controls Caspase-Dependent and -Independent Cell Death Programs Activated by Infection of Monocytes Differentiating to Macrophages
Authors: McCormick A. Louise , Roback Linda , Livingston-Rosanoff Devon , St. Clair Courtney
Published: March 2010

The cellular protease caspase-8 activates extrinsic apoptosis and also functions to promote monocyte-to-macrophage differentiation. Differentiation-induced alterations to antiviral caspase-8-dependent cell death pathways are unclear. Here, we show THP-1 monocyte-to-macrophage differentiation alters the specific cell death pathways activated in response to human cytomegalovirus (HCMV) infection. Employing viruses with mutations in UL36, the gene that encodes theviralinhibitor ofcaspase-8activation (vICA), our data indicate that both caspase-dependent and -independent death pathways are activated in response to infection. Activation of caspase-dependent and -independent cell death responses restricted growth of vICA-deficient viruses, and vICA/pUL36 inhibited either response. Thus, these studies also reveal that the UL36 gene controls a caspase-independent cell death pathway. The impact of caspases on control of antiviral responses differed at early and late stages of macrophage differentiation. Early in differentiation, vICA-deficient virus-induced cell death was dependent on caspases and inhibited by the pan-caspase inhibitor z-VAD(OMe)-fluoromethyl ketone. In contrast, virus-induced death at late times of differentiation was caspase independent. Additional unlabeled and fluorescent inhibitors indicated that caspase-8 promoted death from within infected cells at early but not late stages of differentiation. These data highlight the multifunctional role of vICA/pUL36 as HCMV encounters various antiviral responses during macrophage differentiation.
B Cells and Platelets Harbor Prion Infectivity in the Blood of Deer Infected with Chronic Wasting Disease
Authors: Mathiason Candace K. , Hayes-Klug Jeanette , Hays Sheila A. , Powers Jenny , Osborn David A. , Dahmes Sallie J. , Miller Karl V. , Warren Robert J. , Mason Gary L. , Telling Glenn C. , Young Alan J. , Hoover Edward A.
Published: March 2010

Substantial evidence for prion transmission via blood transfusion exists for many transmissible spongiform encephalopathy (TSE) diseases. Determining which cell phenotype(s) is responsible for trafficking infectivity has important implications for our understanding of the dissemination of prions, as well as their detection and elimination from blood products. We used bioassay studies of native white-tailed deer and transgenic cervidized mice to determine (i) if chronic wasting disease (CWD) blood infectivity is associated with the cellular versus the cell-free/plasma fraction of blood and (ii) in particular if B-cell (MAb 2-104
+
), platelet (CD41/61
+
), or CD14
+
monocyte blood cell phenotypes harbor infectious prions. All four deer transfused with the blood mononuclear cell fraction from CWD
+
donor deer became PrP
CWD
positive by 19 months postinoculation, whereas none of the four deer inoculated with cell-free plasma from the same source developed prion infection. All four of the deer injected with B cells and three of four deer receiving platelets from CWD
+
donor deer became PrP
CWD
positive in as little as 6 months postinoculation, whereas none of the four deer receiving blood CD14
+
monocytes developed evidence of CWD infection (immunohistochemistry and Western blot analysis) after 19 months of observation. Results of the Tg(CerPrP) mouse bioassays mirrored those of the native cervid host. These results indicate that CWD blood infectivity is cell associated and suggest a significant role for B cells and platelets in trafficking CWD infectivity
in vivo
and support earlier tissue-based studies associating putative follicular B cells with PrP
CWD
. Localization of CWD infectivity with leukocyte subpopulations may aid in enhancing the sensitivity of blood-based diagnostic assays for CWD and other TSEs.
The Specificity of TRIM5α-Mediated Restriction Is Influenced by Its Coiled-Coil Domain
Authors: Maillard Pierre V. , Ecco Gabriela , Ortiz Millán , Trono Didier
Published: March 2010

Retroviruses are both powerful evolutionary forces and dangerous threats to genome integrity. As such, they have imposed strong selective pressure on their hosts, notably triggering the emergence of restriction factors, such as TRIM5α, that act as potent barriers to their cross-species transmission. TRIM5α orthologues from different primates have distinct retroviral restriction patterns, largely dictated by the sequence of their C-terminal PRYSPRY domain, which binds the capsid protein of incoming virions. Here, by combining genetic and functional analyses of human and squirrel monkey TRIM5α, we demonstrate that the coiled-coil domain of this protein, thus far essentially known for mediating oligomerization, also conditions the spectrum of antiretroviral activity. Furthermore, we identify three coiled-coil residues responsible for this effect, one of which has been under positive selection during primate evolution, notably in New World monkeys. These results indicate that the PRYSPRY and coiled-coil domains cooperate to determine the specificity of TRIM5α-mediated capture of retroviral capsids, shedding new light on this complex event.
Autographa californica
Multicapsid Nucleopolyhedrovirus Efficiently Infects Sf9 Cells and Transduces Mammalian Cells via Direct Fusion with the Plasma Membrane at Low pH
Authors: Dong Sicong , Wang Manli , Qiu Zhijuan , Deng Fei , Vlak Just M. , Hu Zhihong , Wang Hualin
Published: March 2010

The budded virus (BV) of the
Autographa californica
multicapsid nucleopolyhedrovirus (AcMNPV) infects insect cells and transduces mammalian cells mainly through the endocytosis pathway. However, this study revealed that the treatment of the virus bound to Sf9 cells at low pH could efficiently rescue the infectivity of AcMNPV in the presence of endocytosis pathway inhibitors. A colocalization assay of the major capsid protein VP39 with the early endosome marker EEA1 showed that at low pH, AcMNPV entered Sf9 cells via an endosome-independent pathway. Using a fluorescent probe (R18), we showed that at low pH, the viral nucleocapsid entered Sf9 cells via direct fusion at the cell surface. By using the myosin-specific inhibitor 2,3-butanedione monoxime (BDM) and the microtubule inhibitor nocodazole, the low pH-triggered direct fusion was demonstrated to be dependent on myosin-like proteins and independent of microtubules. The reverse transcription-PCR of the IE1 gene as a marker for viral entry showed that the kinetics of AcMNPV in cells triggered by low pH was similar to that of the normal entry via endocytosis. The low pH-mediated infection assay and VP39 and EEA1 colocalization assay also demonstrated that AcMNPV could efficiently transduce mammalian cells via direct membrane fusion at the cell surface. More importantly, we found that a low-pH trigger could significantly improve the transduction efficiency of AcMNPV in mammalian cells, leading to the potential application of this method when using baculovirus as a vector for heterologous gene expression and for gene therapy.
STATISTICAL CHARACTERISTICS OF SLANT ANGLES IN HANDWRITTEN NUMERAL STRINGS AND EFFECTS OF SLANT CORRECTION ON SEGMENTATION
Authors: SADRI JAVAD , SUEN CHING Y. , BUI TIEN D.
Published: March 2010

A novel and efficient method for correction of slant angles in handwritten numeral strings is proposed. For the first time, the statistical distribution of slant angles in handwritten numerals is investigated and the effects of slant correction on the segmentation of handwritten numeral strings are shown. In our proposed slant correction method, utilizing geometric features, a Component Slant Angle (CSA) is estimated for each connected component independently. A weighted average is then used to compute the String Slant Angle (SSA), which is applied uniformly to correct the slant of all the components in numeral strings. Our experimental results have revealed novel statistics for slant angles of handwritten numeral strings, and also showed that slant correction can significantly improve extraction of segmentation features and segmentation accuracy of touching numerals. Comparison between our slant correction algorithm and similar algorithms in the literature show that our algorithm is more efficient, and on average it has a faster running time.
Morphology and radical reactions of Cu(II) and Co(II) sulfonated phthalocyanines covalently linked to poly(ethyleneamide)
Authors: Ruiz Gustavo T. , Lappin Alexander G. , Ferraudi Guillermo
Published: March 2010

The tetrasulfonated Cu II( tspc )4-, tspc = 4,4′,4″,4‴-phthalocyaninetetrasulfonate, and the trisulfonated Co II( trspc )3-, trspc = n , n ′, n ″-phthalocyanine-trisulfonate and n, n′ and n″ indicate the sulfonated positions of the various isomers, were covalently linked to a polyethyleneimine, M n ~ 10000 and M w ~ 25000, backbone. A fraction of the amine groups in a strand was converted to Cu II pc (- SO 3)3(- SO 2 N <)\3- or Co II pc (- SO 3)2(- SO 2 N <)\2- pendants and the remaining were acetylated. Strands of the polymers, poly(K3CuIItspc) and poly(K2CoIItrspc) , formed spherical bundles with diameters ~1000 nm for poly(K3CuIItspc) and ~100 nm for poly(K2CoIItrspc) . The stability of the bundles is metal-dependent. Poly(K2CoIItrspc) forms the most stable bundles with respect to hydrolytic and redox reactions. ESR and optical spectroscopies as well as reactions of the pendants with pulse radiolytically generated radicals, namely e - sol , C•H2OH, (CH3)2COHC•H2 , CO 2•-, N 3• and SO 4•-, revealed a distribution of pendants between isolated and aggregated forms in a bundle. The reduction of the Cu(II) to Cu(I) in aqueous solutions of the poly(K3CuIItspc) was associated with pendants present in a mostly hydrophobic environment. Unstable phthalocyanine pendants with CoIII -carbon bonds are formed when CoIIpc(-SO3)2(-SO2N<)\2- reacted with C -centered radicals, ( CH3)2COHC•H2 and C•H2OH . The species with CoIII -carbon bonds were precursors to the formation of CoIpc(-SO3)2(-SO2N<)\3- pendants. The redox reactions of the pendants in these polymers are compared with those of K4CuIItspc and K3CoIItrspc .
l-Arginine restores endothelial nitric oxide synthase-coupled activity and attenuates monocrotaline-induced pulmonary artery hypertension in rats
Authors: Ou Zhi-Jun , Wei Wei , Huang Da-De , Luo Wei , Luo Dan , Wang Zhi-Ping , Zhang Xi , Ou Jing-Song
Published: March 2010

l-Arginine can attenuate pulmonary hypertension (PH) by a mechanism that are not fully understood. This study investigated the molecule mechanism of l-arginine attenuating PH. Sprague Dawley rats were treated with monocrotaline (MCT) with or without l-arginine for 3 or 5 wk. Right ventricular systolic pressure (RVSP), right heart hypertrophy, survival rate, pulmonary artery wall thickness, nitric oxide (NO) concentration, and superoxide anion (O2·−) generation in the lung were measured. Expressions of endothelial nitric oxide synthase (eNOS) and heat shock protein 90 (HSP90), phosphorylation of eNOS at Ser1177, and the association of eNOS and HSP90 in the lung were determined by Western blot and immunoprecipitation experiments. MCT increased RVSP, right heart hypertrophy, mortality, pulmonary artery wall thickness, and O2·− generation and decreased eNOS and HSP90 expression and association, phosphorylation of eNOS at Ser1177, and NO production. l-Arginine decreased RVSP, right heart hypertrophy, mortality, O2·− generation, and pulmonary artery wall thickness and increased NO production. l-Arginine increased eNOS expression, phosphorylation of eNOS at Ser1177, and association of eNOS and HSP90 without significantly altering HSP90 expression. l-Arginine may act through three pathways, providing a substrate for NO generation, preserving eNOS expression/phosphorylation, and maintaining the association of eNOS and HSP90, which allows restoration of eNOS activity and coupling activity, to maintain the balance between NO and O2·− and delay the development of PH.
PGC-1α coactivates estrogen-related receptor-α to induce the expression of glucokinase
Authors: Zhu Liu-Luan , Liu Yang , Cui An-Fang , Shao Di , Liang Ji-Chao , Liu Xiao-Jun , Chen Yong , Gupta Nishith , Fang Fu-De , Chang Yong-Sheng
Published: March 2010

Peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) is a key regulator of cellular energy metabolism and regulates processes such as adaptive thermogenesis, hepatic gluconeogenesis, fatty acid oxidation, and mitochondrial biogenesis by coactivating numerous nuclear receptors and transcription factors. Here, we demonstrate the presence of the ERRα binding site in the regulatory sequence of the glucokinase gene and that PGC-1α coactivates ERRα to stimulate the transcription of glucokinase. Simultaneous overexpression of PGC-1α and ERRα potently induced the glucokinase gene expression and its enzymatic activity in primary hepatocytes; however, expression of either PGC-1α or ERRα alone had no significant effect. Electrophoretic mobility shift and chromatin immunoprecipitation assays revealed the interaction of ERRα with the glucokinase promoter. Finally, the knockdown of endogenous ERRα with specific siRNA (siERRα) or pharmacological inhibition of ERRα with XCT790 attenuated insulin-induced glucokinase expression. Taken together, this research identifies glucokinase as a novel target of PGC-1α/ERRα and underscores the regulatory function of ERRα in insulin-dependent enzyme regulation.
In search of autonomic balance: the good, the bad, and the ugly
Authors: Abboud François M.
Published: March 2010

Walter B. Cannon's research on the sympathetic nervous system and neurochemical transmission was pioneering. Wisdom has endowed our body with a powerful autonomic neural regulation of the circulation that provides optimal perfusion of every organ in accordance to its metabolic needs. Exquisite sensors tuned to an optimal internal environment trigger central and peripheral sympathetic and parasympathetic motor neurons and allow desirable and beneficial adjustments to physiologic needs as well as to acute cardiovascular stresses. This short review, presented as The Walter B. Cannon Memorial Award Lecture for 2009, addresses the mechanisms that disrupt sensory signaling and result in a chronic maladjustment of the autonomic neural output that in many cardiovascular diseases results in excessive increases in the risks of dying. The hopes for any reduction of those risks resides in an understanding of the molecular determinants of neuronal signaling.
Estrogen modulates the contribution of neuropeptide Y to baseline hindlimb blood flow control in female Sprague-Dawley rats
Authors: Jackson Dwayne N. , Ellis Christopher G. , Shoemaker J. Kevin
Published: March 2010

The purpose of this study was to determine the role of estrogen in neuropeptide Y (NPY) and Y1 receptor (Y1R)-mediated vascular responses in female rats. Based on earlier work from our laboratory that female rats lacked an NPY contribution to hindlimb vascular conductance relative to males, we tested the hypothesis that estrogen modulates Y1R-mediated hindlimb blood flow control. Thus it was expected that ovariectomy would: 1) increase skeletal muscle Y1R expression, 2) decrease skeletal muscle Y2 receptor (Y2R) expression, 3) decrease peptidase activity, and/or 4) increase overall skeletal muscle NPY concentration. Separate groups of control (CTL), ovariectomized (OVX), and OVX + 17β-estradiol replacement (OVX + E2; 21-day pellet) rats were studied. Animals were anesthetized and given localized hindlimb delivery of BIBP-3226 (Y1R antagonist), while femoral artery blood flow and blood pressure were recorded. Tissue samples from the white and red vastus lateralis muscle were extracted to examine Y1R and Y2R expression, peptidase activity, and NPY concentration. We found that Y1R blockade resulted in increased baseline hindlimb blood flow and vascular conductance in OVX rats, whereas no change was noted in CTL or OVX + E2 groups ( P < 0.05). This enhanced functional effect in the OVX group aligned with greater skeletal muscle Y1R expression in white vastus muscle and a substantial increase in NPY concentration in both white and red vastus muscle compared with CTL and OVX + E2 groups. There was no change in Y2R expression or peptidase activity among the groups. These data support the hypothesis that estrogen blunts Y1R activation in the rat hindlimb through an effect on Y1R expression and NPY concentration.
Heterogeneous stock rats: a new model to study the genetics of renal phenotypes
Authors: Solberg Woods Leah C. , Stelloh Cary , Regner Kevin R. , Schwabe Tiffany , Eisenhauer Jessica , Garrett Michael R.
Published: March 2010

Chronic kidney disease is a growing medical concern, with an estimated 25.6 million people in the United States exhibiting some degree of kidney injury and/or decline in kidney function. Animal models provide great insight into the study of the genetics of complex diseases. In particular, heterogeneous stock (HS) rats represent a unique genetic resource enabling rapid fine-mapping of complex traits. However, they have not been explored as a model to study renal phenotypes. To evaluate the usefulness of HS rats in the genetics of renal traits, a time course evaluation ( weeks 8–40) was performed for several renal phenotypes. As expected, a large degree of variation was seen for most renal traits. By week 24, three (of 40) rats exhibited marked proteinuria that increased gradually until week 40 and ranged from 33.7 to 80.2 mg/24 h. Detailed histological analysis confirmed renal damage in these rats. In addition, several rats consistently exhibited significant hematuria (5/41). Interestingly, these rats were not the same rats that exhibited proteinuria, indicating that susceptibility to different types of kidney injury is likely segregating within the HS population. One HS rat exhibited unilateral renal agenesis (URA), which was accompanied by a significant degree of proteinuria and glomerular and tubulointerstitial injury. The parents of this HS rat were identified and bred further. Additional offspring of this pair were observed to exhibit URA at frequency between 40% and 60%. In summary, these novel data demonstrate that HS rats exhibit variation in proteinuria and other kidney-related traits, confirming that the model harbors susceptibility alleles for kidney injury and providing the basis for further genetic studies.
Proteomic analysis of brush-border membrane vesicles isolated from purified proximal convoluted tubules
Authors: Walmsley Scott J. , Broeckling Corey , Hess Ann , Prenni Jessica , Curthoys Norman P.
Published: March 2010

The renal proximal convoluted tubule is the primary site of water, electrolyte and nutrient reabsorption and of active secretion of selected molecules. Proteins in the apical brush-border membrane facilitate these functions and initiate some of the cellular responses to altered renal physiology. The current study uses two-dimensional liquid chromatography/mass spectrometry to compare brush border membrane vesicles isolated from rat renal cortex (BBMVCTX) and from purified proximal convoluted tubules (BBMVPCT). Both proteomic data and Western blot analysis indicate that the BBMVCTXcontain apical membrane proteins from cortical cells other than the proximal tubule. This heterogeneity was greatly reduced in the BBMVPCT. Proteomic analysis identified 193 proteins common to both samples, 21 proteins unique to BBMVCTX, and 57 proteins unique to BBMVPCT. Spectral counts were used to quantify relative differences in protein abundance. This analysis identified 42 and 50 proteins that are significantly enriched (p values ≤0.001) in the BBMVCTXand BBMVPCT, respectively. These data were validated by measurement of γ-glutamyltranspeptidase activity and by Western blot analysis. The combined results establish that BBMVPCTare primarily derived from the proximal convoluted tubule (S1 and S2 segments), whereas BBMVCTXinclude proteins from the proximal straight tubule (S3 segment). Analysis of functional annotations indicated that BBMVPCTare enriched in mitochondrial proteins and enzymes involved in glucose and organic acid metabolism. Thus the current study reports a detailed proteomic analysis of the brush-border membrane of the rat renal proximal convoluted tubule and provides a database for future hypothesis-driven research.
Generation of Sustained Field Potentials by Gradients of Polarization Within Single Neurons: A Macroscopic Model of Spreading Depression
Authors: Makarova Julia , Makarov Valeri A. , Herreras Oscar
Published: March 2010

Spreading depression (SD) is a pathological wave of depolarization of the neural tissue producing a negative macroscopic field potential ( Vo), used as a marker for diagnostic purposes. The cellular basis of SD and neuronal mechanisms of generation of Vo at the microscopic level are poorly understood. Using a CA1 mathematical model and experimental verification, we examined how transmembrane currents in single cells scale up in the extracellular space shaping Vo. The model includes an array of 17,000 realistically modeled neurons (responsible for generating transmembrane currents) dynamically coupled to a virtual aggregate/extracellular space (responsible for Vo). The SD wave in different tissue bands is simulated by imposing membrane shunts in the corresponding dendritic elements as suggested by experimentally assessed drop in membrane resistance. We show that strong isopotential depolarization of wide domains (as in the main SD phase) produce broad central cancellation of axial and transmembrane currents in single cells. When depolarization is restricted to narrow dendritic domains (as in the late SD phase), the internal cancellation shrinks and the transmembrane current increases. This explains why in the laminated CA1 the Vo is smaller in the main phase of SD, when both dendritic layers are seized, than in the SD tail restricted to an apical band. Moreover, scattering of the neuronal somatas (as in cortical regions) further decreases the aggregate Vo due to the volume averaging. Although mechanistically the Vo associated to SD is similar to customary transient fields, its changes maybe related to spatial factors in single cells rather than cell number or depolarization strength.
Activity-Dependent Release of Endogenous BDNF From Mossy Fibers Evokes a TRPC3 Current and Ca2+Elevations in CA3 Pyramidal Neurons
Authors: Li Yong , Calfa Gaston , Inoue Takafumi , Amaral Michelle D. , Pozzo-Miller Lucas
Published: March 2010

Multiple studies have demonstrated that brain-derived neurotrophic factor (BDNF) is a potent modulator of neuronal structure and function in the hippocampus. However, the majority of studies to date have relied on the application of recombinant BDNF. We herein report that endogenous BDNF, released via theta burst stimulation of mossy fibers (MF), elicits a slowly developing cationic current and intracellular Ca2+elevations in CA3 pyramidal neurons with the same pharmacological profile of the transient receptor potential canonical 3 (TRPC3)-mediated IBDNFactivated in CA1 neurons by brief localized applications of recombinant BDNF. Indeed, sensitivity to both the extracellular BDNF scavenger tropomyosin-related kinase B (TrkB)-IgG and small hairpin interference RNA-mediated TRPC3 channel knockdown confirms the identity of this conductance as such, henceforth-denoted MF- IBDNF. Consistent with such activity-dependent release of BDNF, these MF- IBDNFresponses were insensitive to manipulations of extracellular Zn2+concentration. Brief theta burst stimulation of MFs induced a long-lasting depression in the amplitude of excitatory postsynaptic currents (EPSCs) mediated by both AMPA and N-methyl-d-aspartate (NMDA) receptors without changes in the NMDA receptor/AMPA receptor ratio, suggesting a reduction in neurotransmitter release. This depression of NMDAR-mediated EPSCs required activity-dependent release of endogenous BDNF from MFs and activation of Trk receptors, as it was sensitive to the extracellular BDNF scavenger TrkB-IgG and the tyrosine kinase inhibitor k-252b. These results uncovered the most immediate response to endogenously released—native—BDNF in hippocampal neurons and lend further credence to the relevance of BDNF signaling for synaptic function in the hippocampus.
Gene-family profiling: a normalization-free real-time RT-PCR approach with increased physiological resolution
Authors: Ellefsen Stian , Stensløkken Kåre-Olav
Published: March 2010

Here we present gene-family profiling, an approach for improved real-time RT-PCR analyses. It is based on recently published data, and we argue that it bring solutions to two major problems. First, it is normalization-free and therefore unbiased by variation in normalization agents such as reference gene expression. This strengthens data validity and also increases data resolution, reducing coefficients of variation by ∼48% in our data sets. Second, it includes all members of a particular gene family, treating individual genes as constituting fractions of collective gene-family expression rather than as unrelated entities. Because different family members typically fulfill similar, but complementary roles, this increases the physiological relevance. Gene-family profiling is particularly useful for evaluation of cellular adaptations to physiological challenges and for comparison of properties between different experimental systems such as species, tissues or tissue regions. In addition, it seems suitable for analyses of inherent patterns of gene expression in singular biological samples. In our opinion, the approach is valuable for both research and diagnostic purposes, and should be readily applicable for many studies of gene expression. Its value is likely to increase as science continues to unravel gene function.
Intraosseous Follicular Adenomatoid Odontogenic Tumour—A Case Report
Authors: Durrani Farhan , Singh Royana
Published: March 2010

The adenomatoid odontogenic tumour is a relatively uncommon lesion which mainly affects females in their second decade of life. It exhibits a predilection for the anterior region of the maxilla. The lesion is usually associated with the crown of an embedded tooth, most commonly the maxillary canine. In this paper, we present a case of adenomatoid odontogenic tumor affecting the left maxillary region in a 24-year-old female. The authors also discuss clinical, radiographic, histopathologic, and therapeutic features of the case.
On the Positivity and Zero Crossings of Solutions of Stochastic Volterra Integrodifferential Equations
Authors: Appleby John A. D.
Published: March 2010

We consider the zero crossings and positive solutions
of scalar nonlinear stochastic Volterra integrodifferential equations of Itô type.
In the equations considered, the diffusion coefficient is linear and depends
on the current state, and the drift term is a convolution integral which is in
some sense mean reverting towards the zero equilibrium. The state dependent
restoring force in the integral can be nonlinear. In broad terms, we show that
when the restoring force is of linear or lower order in the neighbourhood of the
equilibrium, or if the kernel decays more slowly than a critical noise‐dependent
rate, then there is a zero crossing almost surely. On the other hand, if the kernel
decays more rapidly than this critical rate, and the restoring force is globally
superlinear, then there is a positive probability that the solution remains of
one sign for all time, given a sufficiently small initial condition. Moreover, the
probability that the solution remains of one sign tends to unity as the initial
condition tends to zero.
A Class Age‐Structured HIV/AIDS Model with Impulsive Drug‐Treatment Strategy
Authors: Liu Helong , Li Lianbing
Published: March 2010

We formulate an HIV/AIDS transmission model
that considers the dependence of HIV/AIDS progress on infection age (the
time since infection), disease age (the time elapsed since the onset), and impulsive antiretroviral treatment. Since no effective vaccine is available for HIV/AIDS, our impulsive disease‐control strategy is targeted at infected individuals (I control). Thus the model only includes infective class and AIDS class: infected population is the state at birth, and AIDS population is not the state at birth. Assuming the theoretical strategy can provide HIV
testing for risk population groups every T years and immediate antiretroviral treatment for HIV‐positive people. The action is approximated by
impulsive differential equations. We demonstrate the effect of the impulsive
drug treatment and show that there exists a globally stable infection‐free
state when the impulsive period Tand drug‐treatment proportion p satisfy R(p, T) < 1. This result shows that the prevention effects can drive HIV/AIDS epidemic towards to elimination.
Recurrent Pneumonia and a Normal Heart: Late Complication after Repair of Hemianomalous Pulmonary Venous Drainage—A Cautionary Tale
Authors: Caruana Maryanne , Grech Victor , Somerville Jane
Published: March 2010

Hemianomalous pulmonary venous drainage with intact atrial septum is a rare congenital anomaly and reports of its surgical repair and the long-term complications related to the correction are only infrequently encountered in the literature. We report the case of a patient with hemianomalous pulmonary venous drainage and intact atrial septum who underwent surgical repair using a pericardial baffle and creation of an “atrial septal defect” aged 15 years. Dyspnoea and recurrent chest infections started 7 months after surgery when he was seen by a respiratory physician without cardiac followup. He presented again aged 28 years with a recurrent pneumonia investigated over 6 weeks and heart pronounced normal from examination and echocardiography. Correct diagnosis was made in Grown Up Congenital Heart (GUCH) clinic stimulating review of data and catheterisation with pulmonary artery angiography which confirmed it. We feel that this case highlights the importance of specialist care and followup for GUCH patients.
Targeting Cancer Cell Metabolism: The Combination of Metformin and 2-Deoxyglucose Induces p53-Dependent Apoptosis in Prostate Cancer Cells
Authors: Ben Sahra Issam , Laurent Kathiane , Giuliano Sandy , Larbret Frédéric , Ponzio Gilles , Gounon Pierre , Le Marchand-Brustel Yannick , Giorgetti-Peraldi Sophie , Cormont Mireille , Bertolotto Corine , Deckert Marcel , Auberger Patrick , Tanti Jean-François , Bost Frédéric
Published: March 2010

Targeting cancer cell metabolism is a new promising strategy to fight cancer. Metformin, a widely used antidiabetic agent, exerts antitumoral and antiproliferative action. In this study, the addition of metformin to 2-deoxyglucose (2DG) inhibited mitochondrial respiration and glycolysis in prostate cancer cells leading to a severe depletion in ATP. The combination of the two drugs was much more harmful for cancer cells than the treatment with metformin or 2DG alone, leading to 96% inhibition of cell viability in LNCaP prostate cancer cells. In contrast, a moderate effect on cell viability was observed in normal prostate epithelial cells. At the cellular level, the combination of metformin and 2DG induced p53-dependent apoptosis via the energy sensor pathway AMP kinase, and the reexpression of a functional p53 in p53-deficient prostate cancer cells restored caspase-3 activity. In addition to apoptosis, the combination of metformin and 2DG arrested prostate cancer cells in G2-M. This G2-M arrest was independent of p53 and correlated with a stronger decrease in cell viability than obtained with either drug. Finally, metformin inhibited 2DG-induced autophagy, decreased beclin 1 expression, and triggered a switch from a survival process to cell death. Our study reinforces the growing interest of metabolic perturbators in cancer therapy and highlights the potential use of the combination of metformin and 2DG as an anticancerous treatment. Cancer Res; 70(6); 2465–75
Local Radiation Therapy Inhibits Tumor Growth through the Generation of Tumor-Specific CTL: Its Potentiation by Combination with Th1 Cell Therapy
Authors: Takeshima Tsuguhide , Chamoto Kenji , Wakita Daiko , Ohkuri Takayuki , Togashi Yuji , Shirato Hiroki , Kitamura Hidemitsu , Nishimura Takashi
Published: March 2010

Radiation therapy is one of the primary treatment modalities for cancer along with chemotherapy and surgical therapy. The main mechanism of the tumor reduction after irradiation has been considered to be damage to the tumor DNA. However, we found that tumor-specific CTL, which were induced in the draining lymph nodes (DLN) and tumor tissue of tumor-bearing mice, play a crucial role in the inhibition of tumor growth by radiation. Indeed, the therapeutic effect of irradiation was almost completely abolished in tumor-bearing mice by depleting CD8+ T cells through anti-CD8 monoclonal antibody administration. In mice whose DLN were surgically ablated or genetically defective (Aly/Aly mice), the generation of tetramer+ tumor-specific CTL at the tumor site was greatly reduced in parallel with the attenuation of the radiation-induced therapeutic effect against the tumor. This indicates that DLN are essential for the activation and accumulation of radiation-induced CTL, which are essential for inhibition of the tumor. A combined therapy of local radiation with Th1 cell therapy augmented the generation of tumor-specific CTL at the tumor site and induced a complete regression of the tumor, although radiation therapy alone did not exhibit such a pronounced therapeutic effect. Thus, we conclude that the combination treatment of local radiation therapy and Th1 cell therapy is a rational strategy to augment antitumor activity mediated by tumor-specific CTL. Cancer Res; 70(7); 2697–706
Matrix Metalloproteinases Contribute Distinct Roles in Neuroendocrine Prostate Carcinogenesis, Metastasis, and Angiogenesis Progression
Authors: Littlepage Laurie E. , Sternlicht Mark D. , Rougier Nathalie , Phillips Joanna , Gallo Eugenio , Yu Ying , Williams Kurt , Brenot Audrey , Gordon Jeffrey I. , Werb Zena
Published: March 2010

Prostate cancer is the leading form of cancer in men. Prostate tumors often contain neuroendocrine differentiation, which correlates with androgen-independent progression and poor prognosis. Matrix metalloproteinases (MMP), a family of enzymes that remodel the microenvironment, are associated with tumorigenesis and metastasis. To evaluate MMPs during metastatic prostatic neuroendocrine cancer development, we used transgenic mice expressing SV40 large T antigen in their prostatic neuroendocrine cells, under the control of transcriptional regulatory elements from the mouse cryptdin-2 gene (CR2-TAg). These mice have a stereotypical pattern of tumorigenesis and metastasis. MMP-2, MMP-7, and MMP-9 activities increased concurrently with the transition to invasive metastatic carcinoma, but they were expressed in different prostatic cell types: stromal, luminal epithelium, and macrophages, respectively. CR2-TAg mice treated with AG3340/Prinomastat, an MMP inhibitor that blocks activity of MMP-2, MMP-9, MMP-13, and MMP-14, had reduced tumor burden. CR2-TAg animals were crossed to mice homozygous for null alleles of MMP-2, MMP-7, or MMP-9 genes. At 24 weeks CR2-TAg; MMP-2−/− mice showed reduced tumor burden, prolonged survival, decreased lung metastasis, and decreased blood vessel density, whereas deficiencies in MMP-7 or MMP-9 did not influence tumor growth or survival. Mice deficient for MMP-7 had reduced endothelial area coverage and decreased vessel size, and mice lacking MMP-9 had increased numbers of invasive foci and increased perivascular invasion, as well as decreased tumor blood vessel size. Together, these results suggest distinct contributions by MMPs to the progression of aggressive prostate tumor and to helping tumors cleverly find alternative routes to malignant progression. Cancer Res; 70(6); 2224–34
Randomized Phase II Multicenter Trial of Two Schedules of Lapatinib as First- or Second-Line Monotherapy in Patients with Advanced or Metastatic Non–Small Cell Lung Cancer
Authors: Ross Helen J. , Blumenschein, George R. , Aisner Joseph , Damjanov Nevena , Dowlati Afshin , Garst Jennifer , Rigas James R. , Smylie Michael , Hassani Habib , Allen Kimberly E. , Leopold Lance , Zaks Tal Z. , Shepherd Frances A.
Published: March 2010

Purpose: This randomized phase II study was initially designed to test the activity of two dose schedules of lapatinib (GW572016H), an oral, reversible, dual tyrosine kinase inhibitor of epidermal growth factor receptor (EGFR) and human EGFR-2 (HER2/neu; HER2), in chemotherapy-naive patients with non–small cell lung cancer (NSCLC); it was later amended to target patients with bronchioloalveolar carcinoma or no smoking history.

Experimental Design: Patients with good performance status and recurrent or metastatic NSCLC were randomized to lapatinib (orally, 1,500 mg once daily or 500 mg twice daily) until progression or intolerance. Patients could have had a maximum of one prior systemic therapy (chemotherapy or biological therapy) for NSCLC. Safety and activity were assessed every 4 and 8 weeks, respectively. Tumors were analyzed for EGFR and HER2 mutations and/or amplifications.

Results: Of 75 patients in the nontargeted population, 1 (1.3%) had partial response and 16 (21%) had stable disease of ≥24 weeks. No complete or partial responses were observed in 56 patients in the targeted population; 14 (25%) had stable disease of ≥24 weeks. No responses were seen in three patients with EGFR mutations and five with EGFR gene amplification. No mutations in HER2 were found. One of two patients with HER2 amplification had a 51% decrease in tumor size; however, this response was unconfirmed. The most common adverse events were grade 1 or 2 diarrhea, rash, fatigue, nausea, and anorexia. Adverse events were similar across dosing regimens.

Conclusions: Lapatinib was well tolerated, with no notable difference in toxicity between treatment groups. Lapatinib monotherapy did not induce a significant number of tumor regressions in NSCLC. Further studies may be warranted to determine whether lapatinib is active in combination with other agents in the treatment of NSCLC. Clin Cancer Res; 16(6); 1938–49
Overexpression of Smoothened Activates the Sonic Hedgehog Signaling Pathway in Pancreatic Cancer–Associated Fibroblasts
Authors: Walter Kimberly , Omura Noriyuki , Hong Seung-Mo , Griffith Margaret , Vincent Audrey , Borges Michael , Goggins Michael
Published: March 2010

Purpose: Accumulating evidence suggests that cancer-associated stromal fibroblasts (CAF) contribute to tumor growth by actively communicating with cancer cells. Our aim is to identify signaling pathways involved in tumor-stromal cell interactions in human pancreatic cancer.

Experimental Design: We established primary fibroblast cultures from human pancreatic adenocarcinomas and nonneoplastic pancreas tissues. To identify differentially expressed genes in CAFs, we did gene expression profiling of human pancreatic CAFs and nonneoplastic pancreatic fibroblasts.

Results: The Hedgehog receptor Smoothened (SMO) was upregulated in CAFs relative to control fibroblasts. CAFs expressing SMO could transduce the Sonic hedgehog signal to activate Gli1 expression, and small interfering RNA knockdown of SMO blocked the induction of Gli1 in these cells. Stromal fibroblasts of human primary pancreatic adenocarcinomas overexpressed Smo compared with normal pancreatic fibroblasts.

Conclusions: These findings implicate overexpression of Smo as a mechanism for the activation of Hedgehog signaling in human pancreatic CAFs and suggest that stromal cells may be a therapeutic target for Smo antagonists in pancreatic cancer. Clin Cancer Res; 16(6); 1781–9
An Overview of the Optimal Planning, Design, and Conduct of Phase I Studies of New Therapeutics
Authors: LoRusso Patricia M. , Boerner Scott A. , Seymour Lesley
Published: March 2010

Phase I clinical trials represent the first step in bringing promising new treatments from the laboratory to the clinic. Although the importance of phase I clinical trials is widely recognized, there is currently no consensus among the scientific, medical, and statistical communities on how best to do these studies in humans. With the advent of targeted therapies, it has become evident that we need to tailor the design of phase I studies for the particular drug class under investigation and any endpoints that are being defined.

The National Cancer Institute (NCI) Investigational Drug Steering Committee (IDSC) provides broad external scientific and clinical input on the design and prioritization of early-phase clinical trials with agents for which the NCI Cancer Therapy Evaluation Program (CTEP) holds an Investigational New Drug (IND) application through the U.S. Food and Drug Administration (FDA). The IDSC has formed a number of task forces and working groups, including the Clinical Trial Design Task Force and the Biomarker Working Group, many with membership from within the IDSC as well as external experts, including participants from academia, the pharmaceutical industry, and regulatory authorities.

The Clinical Trials Design Taskforce sponsored a Phase I Workshop with the primary goal being to develop consensus recommendations for the optimal design of phase I studies. The primary focus included (1) efficient trial designs, (2) phase I drug combinations, and (3) appropriate statistical and correlative endpoints.

In this CCR Focus series, articles summarize key aspects and recommendations on phase I studies (including combination trials), such as design, use of biomarkers, the European Union and Japanese perspectives on design, requirements for first-in-human and other phase I studies, and ensuring regulatory and International Conference on Harmonization (ICH) compliance. A final article summarizes recommendations for the design and conduct of phase II studies. Clin Cancer Res; 16(6); 1710–8
Guidelines for the Development and Incorporation of Biomarker Studies in Early Clinical Trials of Novel Agents
Authors: Dancey Janet E. , Dobbin Kevin K. , Groshen Susan , Jessup J. Milburn , Hruszkewycz Andrew H. , Koehler Maria , Parchment Ralph , Ratain Mark J. , Shankar Lalitha K. , Stadler Walter M. , True Lawrence D. , Gravell Amy , Grever Michael R.
Published: March 2010

The National Cancer Institute (NCI) Investigational Drug Steering Committee (IDSC) charged the Biomarker Task Force to develop recommendations to improve the decisions about incorporation of biomarker studies in early investigational drug trials. The Task Force members reviewed biomarker trials, the peer-reviewed literature, NCI and U.S. Food and Drug Administration (FDA) guidance documents, and conducted a survey of investigators to determine practices and challenges to executing biomarker studies in clinical trials of new drugs in early development. This document provides standard definitions and categories of biomarkers, and lists recommendations to sponsors and investigators for biomarker incorporation into such trials. Our recommendations for sponsors focus on the identification and prioritization of biomarkers and assays, the coordination of activities for the development and use of assays, and for operational activities. We also provide recommendations for investigators developing clinical trials with biomarker studies for scientific rationale, assay criteria, trial design, and analysis. The incorporation of biomarker studies into early drug trials is complex. Thus the decision to proceed with studies of biomarkers should be based on balancing the strength of science, assay robustness, feasibility, and resources with the burden of proper sample collection on the patient and potential impact of the results on drug development. The Task Force provides these guidelines in the hopes that improvements in biomarker studies will enhance the efficiency of investigational drug development. Clin Cancer Res; 16(6); 1745–55
Performing Phase I Clinical Trials of Anticancer Agents: Perspectives from within the European Union and Japan
Authors: Forster Martin D. , Saijo Nagahiro , Seymour Lesley , Calvert Hilary
Published: March 2010

Drug discovery and early clinical development is an international endeavor, conducted in partnership between commercial entities such as biotechnology and pharmaceutical companies and academic investigators. Although once considered quite disparate, early clinical trials requirements and conduct are largely harmonized between the European Union, Japan, and the United States, increasing the opportunities for productive commercial-academic collaborations. Clin Cancer Res; 16(6); 1737–44