bluuebunny/crossref_metadata_2025 huggingface dataset:
Multilocus Sequence Analysis and
rpoB
Sequencing of Mycobacterium abscessus (Sensu Lato) Strains
Authors: Macheras Edouard , Roux Anne-Laure , Bastian Sylvaine , Leão Sylvia Cardoso , Palaci Moises , Sivadon-Tardy Valérie , Gutierrez Cristina , Richter Elvira , Rüsch-Gerdes Sabine , Pfyffer Gaby , Bodmer Thomas , Cambau Emmanuelle , Gaillard Jean-Louis , Heym Beate
Published: November 2010

Mycobacterium abscessus
,
Mycobacterium bolletii
, and
Mycobacterium massiliense
(
Mycobacterium abscessus
sensu lato) are closely related species that currently are identified by the sequencing of the
rpoB
gene. However, recent studies show that
rpoB
sequencing alone is insufficient to discriminate between these species, and some authors have questioned their current taxonomic classification. We studied here a large collection of
M. abscessus
(sensu lato) strains by partial
rpoB
sequencing (752 bp) and multilocus sequence analysis (MLSA). The final MLSA scheme developed was based on the partial sequences of eight housekeeping genes:
argH
,
cya
,
glpK
,
gnd
,
murC
,
pgm
,
pta
, and
purH
. The strains studied included the three type strains (
M. abscessus
CIP 104536
T
,
M. massiliense
CIP 108297
T
, and
M. bolletii
CIP 108541
T
) and 120 isolates recovered between 1997 and 2007 in France, Germany, Switzerland, and Brazil. The
rpoB
phylogenetic tree confirmed the existence of three main clusters, each comprising the type strain of one species. However, divergence values between the
M. massiliense
and
M. bolletii
clusters all were below 3% and between the
M. abscessus
and
M. massiliense
clusters were from 2.66 to 3.59%. The tree produced using the concatenated MLSA gene sequences (4,071 bp) also showed three main clusters, each comprising the type strain of one species. The
M. abscessus
cluster had a bootstrap value of 100% and was mostly compact. Bootstrap values for the
M. massiliense
and
M. bolletii
branches were much lower (71 and 61%, respectively), with the
M. massiliense
cluster having a fuzzy aspect. Mean (range) divergence values were 2.17% (1.13 to 2.58%) between the
M. abscessus
and
M. massiliense
clusters, 2.37% (1.5 to 2.85%) between the
M. abscessus
and
M. bolletii
clusters, and 2.28% (0.86 to 2.68%) between the
M. massiliense
and
M. bolletii
clusters. Adding the
rpoB
sequence to the MLSA-concatenated sequence (total sequence, 4,823 bp) had little effect on the clustering of strains. We found 10/120 (8.3%) isolates for which the concatenated MLSA gene sequence and
rpoB
sequence were discordant (e.g.,
M. massiliense
MLSA sequence and
M. abscessus rpoB
sequence), suggesting the intergroup lateral transfers of
rpoB
. In conclusion, our study strongly supports the recent proposal that
M. abscessus
,
M. massiliense
, and
M. bolletii
should constitute a single species. Our findings also indicate that there has been a horizontal transfer of
rpoB
sequences between these subgroups, precluding the use of
rpoB
sequencing alone for the accurate identification of the two proposed
M. abscessus
subspecies.
Hepatitis B and C Virus Infections in the Three Pygmy Groups in Cameroon
Authors: Foupouapouognigni Yacouba , Sadeuh Mba Serge Alain , à Betsem Edouard Betsem , Rousset Dominique , Froment Alain , Gessain Antoine , Njouom Richard
Published: November 2010

A high (11.8%) level of hepatitis B virus (HBV) infection was found among 524 Pygmies in Cameroon, whereas the extent of hepatitis C virus (HCV) infection in the same population was low (0.6%). Phylogenetic analyses showed cocirculation of two HBV genotypes, HBV-A3 and -E. Taken together, our results suggest different epidemiological scenarios concerning HBV and HCV infections in this population.
Role of the CM2 Protein in the Influenza C Virus Replication Cycle
Authors: Furukawa Takatoshi , Muraki Yasushi , Noda Takeshi , Takashita Emi , Sho Ri , Sugawara Kanetsu , Matsuzaki Yoko , Shimotai Yoshitaka , Hongo Seiji
Published: November 2010

CM2 is the second membrane protein of influenza C virus. Although its biochemical characteristics, coding strategy, and properties as an ion channel have been extensively studied, the role(s) of CM2 in the virus replication cycle remains to be clarified. In order to elucidate this role, in the present study we generated CM2-deficient influenza C virus-like particles (VLPs) and examined the VLP-producing 293T cells, VLPs, and VLP-infected HMV-II cells. Quantification of viral RNA (vRNA) in the VLPs by real-time PCR revealed that the CM2-deficient VLPs contain approximately one-third of the vRNA found in wild-type VLPs although no significant differences were detected in the expression levels of viral components in VLP-producing cells or in the number and morphology of the generated VLPs. This finding suggests that CM2 is involved in the genome packaging process into VLPs. Furthermore, HMV-II cells infected with CM2-deficient VLPs exhibited significantly reduced reporter gene expression. Although CM2-deficient VLPs could be internalized into HMV-II cells as efficiently as wild-type VLPs, a smaller amount of vRNA was detected in the nuclear fraction of CM2-deficient VLP-infected cells than in that of wild-type VLP-infected cells, suggesting that the uncoating process of the CM2-deficient VLPs in the infected cells did not proceed in an appropriate manner. Taken together, the data obtained in the present study indicate that CM2 has a potential role in the genome packaging and uncoating processes of the virus replication cycle.
Rabies Virus Expressing Dendritic Cell-Activating Molecules Enhances the Innate and Adaptive Immune Response to Vaccination
Authors: Wen Yongjun , Wang Hualei , Wu Hua , Yang Fuhe , Tripp Ralph A. , Hogan Robert J. , Fu Zhen F.
Published: November 2010

Our previous studies indicated that recruitment and/or activation of dendritic cells (DCs) is important in enhancing the protective immune responses against rabies virus (RABV) (L. Zhao, H. Toriumi, H. Wang, Y. Kuang, X. Guo, K. Morimoto, and Z. F. Fu, J. Virol. 84:9642-9648). To address the importance of DC activation for RABV vaccine efficacy, the genes for several DC recruitment and/or activation molecules, e.g., granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage-derived chemokine (MDC), and macrophage inflammatory protein 1α (MIP-1α), were individually cloned into RABV. The ability of these recombinant viruses to activate DCs was determinedin vitroandin vivo. Infection of mouse bone marrow-derived DCs with each of the recombinant viruses resulted in DC activation, as shown by increased surface expression of CD11c and CD86 as well as an increased level of alpha interferon (IFN-α) production compared to levels observed after infection with the parent virus. Intramuscular infection of mice with each of the viruses recruited and/or activated more DCs and B cells in the periphery than infection with the parent virus, leading to the production of higher levels of virus-neutralizing antibodies. Furthermore, a single immunization with recombinant RABV expressing GM-CSF or MDC protected significantly more mice against intracerebral challenge with virulent RABV than did immunization with the parental virus. Yet, these viruses did not show more virulence than the parent virus, since direct intracerebral inoculation with each virus at up to 1 × 107fluorescent focus units each did not induce any overt clinic symptom, such as abnormal behavior, or any neurological signs. Together, these data indicate that recombinant RABVs expressing these molecules activate/recruit DCs and enhance protective immune responses.
The Activator Protein 1 Binding Motifs within the Human Cytomegalovirus Major Immediate-Early Enhancer Are Functionally Redundant and Act in a Cooperative Manner with the NF-κB Sites during Acute Infection
Authors: Isern Elena , Gustems Montse , Messerle Martin , Borst Eva , Ghazal Peter , Angulo Ana
Published: November 2010

Human cytomegalovirus (HCMV) infection causes a rapid induction of c-Fos and c-Jun, the major subunits of activator protein 1 (AP-1), which in turn have been postulated to activate the viral immediate-early (IE) genes. Accordingly, the major IE promoter (MIEP) enhancer, a critical control region for initiating lytic HCMV infection and reactivation from the latent state, contains one well-characterized AP-1 site and a second candidate interaction site. In this study we explored the role of these AP-1 elements in the context of the infection. We first show that the distal candidate AP-1 motif binds c-Fos/c-Jun heterodimers (AP-1 complex) and confers c-Fos/c-Jun-mediated activity to a core promoter. Site-directed mutagenesis studies indicate that both AP-1 response elements are critical for 12-
O
-tetradecanoylphorbol-13-acetate (TPA)-enhanced MIEP activity in transient-transfection assays. In marked contrast to the results obtained with the isolated promoter, disruption of the AP-1 recognition sites of the MIEP in the context of the infectious HCMV genome has no significant influence on the expression of the MIE protein IE1 or viral replication in different cell types. Moreover, a chimeric murine CMV driven by the HCMV MIEP (hMCMV-ES) with the two AP-1 binding sites mutated is not compromised in virulence, is able to grow and disseminate to different organs of the newborn mice as efficiently as the parental virus, and is competent in reactivation. We show, however, that combined inactivation of the enhancer AP-1 and NF-κB recognition sites leads to an attenuation of the hMCMV-ES in the neonatal murine infection model, not observed when each single element is abolished. Altogether, these results underline the functional redundancy of the MIEP elements, highlighting the plasticity of this region, which probably evolved to ensure maximal transcriptional performance across many diverse environments.
Coordinate Roles of Gag and RNA Helicase A in Promoting the Annealing of \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(\mathrm{tRNA}_{3}^{\mathrm{Lys}}\) \end{document} to HIV-1 RNA
Authors: Xing Li , Liang Chen , Kleiman Lawrence
Published: November 2010

RNA helicase A (RHA) has been shown to promote HIV-1 replication at both the translation and reverse transcription stages. A prerequisite step for reverse transcription involves the annealing of

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, the primer for reverse transcription, to HIV-1 RNA.

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annealing is a multistep process that is initially facilitated by Gag prior to viral protein processing. Herein, we report that RHA promotes this annealing through increasing both the quantity of

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annealed by Gag and the ability of

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to prime the initiation of reverse transcription. This improved annealing is the result of an altered viral RNA conformation produced by the coordinate action of Gag and RHA. Since RHA has been reported to promote the translation of unspliced viral RNA to Gag protein, our observations suggest that the conformational change in viral RNA induced by RHA and newly produced Gag may help facilitate the switch in viral RNA from a translational mode to one facilitating

\batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(tRNA_{3}^{Lys}\) \end{document}

annealing.
Factors predicting incorrect use of restraints by children travelling in cars: a cluster randomised observational study
Authors: Bilston Lynne E , Du Wei , Brown Julie
Published: November 2010

Background

The benefits of correctly using size-appropriate restraints for children travelling in cars are well established, and considerable research has focused on the determinants of appropriate restraint choice. There are few studies of the factors associated with incorrect use.

Objective

To determine predictors of incorrect restraint use by child occupants and how these differ from inappropriate use.

Methods

A stratified multistage cluster sample of child occupants aged 0–10 years in New South Wales, Australia was used. Observation of restraint type and inspection of correctness of use was performed on arrival at schools, childcare centres, and child health clinics. An interview was conducted with the driver. Logistic regression was performed to estimate the effects of parental, family, and other characteristics on the likelihood of moderate or serious incorrect restraint use for restraint classes (rear/forward facing, booster, and seatbelt).

Results

Significant factors varied depending on age and restraint class. Older child restraint users (OR per year of age 0.27, 95% CI 0.07 to 0.98) and seatbelt users (OR per year of age 0.54, 95% CI 0.45 to 0.64) were less likely to be incorrectly using their restraints than younger users. Child restraint and booster users from non-English speaking families were more likely to be incorrectly using their restraints. Having more children in the car appeared to reduce incorrect use for booster (OR 0.18, 95% CI 0.06 to 0.57) and seatbelt users (OR 0.39, 95% CI 0.16 to 0.93).

Conclusions

There is a need to reduce incorrect restraint use by both education and improved restraint design. Education aimed at reducing incorrect use may need to be targeted differently to appropriate use programmes, as the predictive factors differ.
Immunohistochemical assessment for Cdx2 expression in the Barrett metaplasia–dysplasia–adenocarcinoma sequence
Authors: Hayes Stephen , Ahmed Sajad , Clark Peter
Published: November 2010

Background

Barrett oesophagus (BO) occurs as a consequence of prolonged gastro-oesophageal reflux and it may precede the development of oesophageal adenocarcinoma.

Aims

The aim of this study was to accurately assess Cdx2 expression in the oesophageal metaplasia–dysplasia–adenocarcinoma sequence.

Method

The expression of the transcription factor Cdx2 in Barrett metaplasia, high-grade glandular dysplasia and adenocarcinoma was investigated using immunohistochemistry.

Results

The results confirmed previous immunohistochemistry and PCR-based investigations, indicating that Cdx2 was expressed by intestinal metaplasia in the oesophagus. In addition, upregulation followed by linear downregulation of Cdx2 expression through the oesophageal metaplasia–dysplasia–adenocarcinoma sequence was demonstrated for the first time.

Conclusion

Such downregulation of Cdx2 expression could be in keeping with a role as a tumour suppressor gene, but a simpler explanation would be downregulation of a differentiation marker.
Loss of vision
Authors: Lueck Christian J
Published: November 2010

Visual loss is not uncommon and many patients end up seeing neurologists because of it. There is a long list of possible causes but in most patients visual loss is associated with visual field loss. This means that for practical purposes the differential diagnosis can usually be narrowed down to a manageable shortlist by consideration of where in the visual pathway the lesion is likely to be, along with the time course of the visual loss. This article provides a practical approach to the diagnosis and appropriate investigation of such patients, dividing them into four groups: those in whom vision is lost transiently, acutely, subacutely (ie, days to weeks) and over a longer time frame (months to years). In addition, there is a discussion of those patients in whom visual loss is not obviously accompanied by any visual field loss.
ON THE POLYTOPES OF BELIEF AND PLAUSIBILITY FUNCTIONS
Authors: MIRANDA P. , COMBARRO E. F.
Published: November 2010

In this paper we study some properties of the polytope of belief functions on a finite referential. These properties can be used in the problem of identification of a belief function from sample data. More concretely, we study the set of isometries, the set of invariant measures and the adjacency structure. From these results, we prove that the polytope of belief functions is not an order polytope if the referential has more than two elements. Similar results are obtained for plausibility functions.
Low-affinity CCK-A receptors are coexpressed with leptin receptors in rat nodose ganglia: implications for leptin as a regulator of short-term satiety
Authors: Li Ying , Wu Xiaoyin , Zhou Shiyi , Owyang Chung
Published: November 2010

The paradigm for the control of feeding behavior has changed significantly. Research has shown that leptin, in the presence of CCK, may mediate the control of short-term food intake. This interaction between CCK and leptin occurs at the vagus nerve. In the present study, we aimed to characterize the interaction between CCK and leptin in the vagal primary afferent neurons. Single neuronal discharges of vagal primary afferent neurons innervating the gastrointestinal tract were recorded from rat nodose ganglia. Three groups of nodose ganglia neurons were identified: group 1 responded to CCK-8 but not leptin; group 2 responded to leptin but not CCK-8; group 3 responded to high-dose CCK-8 and leptin. In fact, the neurons in group 3 showed CCK-8 and leptin potentiation, and they responded to gastric distention. To identify the CCK-A receptor (CCKAR) affinity states that colocalize with the leptin receptor OB-Rb, we used CCK-JMV-180, a high-affinity CCKAR agonist and low-affinity CCKAR antagonist. As expected, immunohistochemical studies showed that CCK-8 administration significantly potentiated the increase in the number of c-Fos-positive neurons stimulated by leptin in vagal nodose ganglia. Administration of CCK-JMV-180 eliminated the synergistic interaction between CCK-8 and leptin. We conclude that both low- and high-affinity CCKAR are expressed in nodose ganglia. Many nodose neurons bearing low-affinity CCKAR express OB-Rb. These neurons also respond to mechanical distention. An interaction between CCKAR and OB-Rb in these neurons likely facilitates leptin mediation of short-term satiety.
Dietary sodium modulates the interaction between efferent and afferent renal nerve activity by altering activation of α2-adrenoceptors on renal sensory nerves
Authors: Kopp Ulla C. , Cicha Michael Z. , Smith Lori A. , Ruohonen Saku , Scheinin Mika , Fritz Nicolas , Hökfelt Tomas
Published: November 2010

Activation of efferent renal sympathetic nerve activity (ERSNA) increases afferent renal nerve activity (ARNA), which then reflexively decreases ERSNA via activation of the renorenal reflexes to maintain low ERSNA. The ERSNA-ARNA interaction is mediated by norepinephrine (NE) that increases and decreases ARNA by activation of renal α1-and α2-adrenoceptors (AR), respectively. The ERSNA-induced increases in ARNA are suppressed during a low-sodium (2,470 ± 770% s) and enhanced during a high-sodium diet (5,670 ± 1,260% s). We examined the role of α2-AR in modulating the responsiveness of renal sensory nerves during low- and high-sodium diets. Immunohistochemical analysis suggested the presence of α2A-AR and α2C-AR subtypes on renal sensory nerves. During the low-sodium diet, renal pelvic administration of the α2-AR antagonist rauwolscine or the AT1 receptor antagonist losartan alone failed to alter the ARNA responses to reflex increases in ERSNA. Likewise, renal pelvic release of substance P produced by 250 pM NE (from 8.0 ± 1.3 to 8.5 ± 1.6 pg/min) was not affected by rauwolscine or losartan alone. However, rauwolscine+losartan enhanced the ARNA responses to reflex increases in ERSNA (4,680 ± 1,240%·s), and renal pelvic release of substance P by 250 pM NE, from 8.3 ± 0.6 to 14.2 ± 0.8 pg/min. During a high-sodium diet, rauwolscine had no effect on the ARNA response to reflex increases in ERSNA or renal pelvic release of substance P produced by NE. Losartan was not examined because of low endogenous ANG II levels in renal pelvic tissue during a high-sodium diet. Increased activation of α2-AR contributes to the reduced interaction between ERSNA and ARNA during low-sodium intake, whereas no/minimal activation of α2-AR contributes to the enhanced ERSNA-ARNA interaction under conditions of high sodium intake.
High doses of vitamin E improve mitochondrial dysfunction in rat hippocampus and frontal cortex upon aging
Authors: Navarro Ana , Bandez Manuel J. , Lopez-Cepero Jose M. , Gómez Carmen , Boveris Alberto
Published: November 2010

Rat aging from 4 to 12 mo was accompanied by hippocampus and frontal cortex mitochondrial dysfunction, with decreases of 23 to 53% in tissue and mitochondrial respiration and in the activities of complexes I and IV and of mitochondrial nitric oxide synthase (mtNOS) ( P < 0.02). In aged rats, the two brain areas showed mitochondria with higher content (35–78%) of oxidation products of phospholipids and proteins and with higher (59–95%) rates of O2−and H2O2production ( P < 0.02). Dietary supplementation with vitamin E (2.0 or 5.0 g/kg of food) from 9 to 12 mo of rat age, restored in a dose-dependent manner, the decreases in tissue and mitochondrial respiration (to 90–96%) and complexes I and IV and mtNOS activities (to 86–88%) of the values of 4-mo-old rats ( P < 0.02). Vitamin E prevented, by 73–80%, the increases in oxidation products, and by 62–68%, the increases in O2−and H2O2production ( P < 0.05). High resolution histochemistry of cytochrome oxidase in the hippocampal CA1 region showed higher staining in vitamin E-treated rats than in control animals. Aging decreased (19%) hippocampus mitochondrial mass, an effect that was restored by vitamin E. High doses of vitamin E seem to sustain mitochondrial biogenesis in synaptic areas.
Electrostimulation during hindlimb unloading modulates PI3K-AKT downstream targets without preventing soleus atrophy and restores slow phenotype through ERK
Authors: Dupont Erwan , Cieniewski-Bernard Caroline , Bastide Bruno , Stevens Laurence
Published: November 2010

Our aim was to analyze the role of phosphatidylinositol 3-kinase (PI3K)-AKT and MAPK signaling pathways in the regulation of muscle mass and slow-to-fast phenotype transition during hindlimb unloading (HU). For that purpose, we studied, in rat slow soleus and fast extensor digitorum longus muscles, the time course of anabolic PI3K-AKT-mammalian target of rapamycin, catabolic PI3K-AKT-forkhead box O (FOXO), and MAPK signaling pathway activation after 7, 14, and 28 days of HU. Moreover, we performed chronic low-frequency soleus electrostimulation during HU to maintain exclusively contractile phenotype and so to determine more precisely the role of these signaling pathways in the modulation of muscle mass. HU induced a downregulation of the anabolic AKT, mammalian target of rapamycin, 70-kDa ribosomal protein S6 kinase, 4E-binding protein 1, and glycogen synthase kinase-3β targets, and an upregulation of the catabolic FOXO1 and muscle-specific RING finger protein-1 targets correlated with soleus muscle atrophy. Unexpectedly, soleus electrostimulation maintained 70-kDa ribosomal protein S6 kinase, 4E-binding protein 1, FOXO1, and muscle-specific RING finger protein-1 to control levels, but failed to reduce muscle atrophy. HU decreased ERK phosphorylation, while electrostimulation enabled the maintenance of ERK phosphorylation similar to control level. Moreover, slow-to-fast myosin heavy chain phenotype transition and upregulated glycolytic metabolism were prevented by soleus electrostimulation during HU. Taken together, our data demonstrated that the processes responsible for gradual disuse muscle plasticity in HU conditions involved both PI3-AKT and MAPK pathways. Moreover, electrostimulation during HU restored PI3K-AKT activation without counteracting soleus atrophy, suggesting the involvement of other signaling pathways. Finally, electrostimulation maintained initial contractile and metabolism properties in parallel to ERK activation, reinforcing the idea of a predominant role of ERK in the regulation of muscle slow phenotype.
ELISA for Aging Biomarkers Induced by Telomere Dysfunction in Human Plasma
Authors: Jiang Hong , Chen Wenqing , Qu Lihui , Chen Ying , He Qiang , Wang Huiping , Wu Jianyong , Shou Zhangfei , Ju Zhenyu , Chen Jianghua
Published: November 2010

Background. We identified cathelicidin related antimicrobial protein (CRAMP) secreted from telomere dysfunctional bone marrow cells of late generation telomerase knockout mice (G4mTerc−/−), increased in blood and various tissues. It can represented human aging and disease. The main aim of this study is to investigate the sensitive direct enzyme-linked immunosorbent assay (ELISA) method to analyze the human aging and disease in plasma and the detailed methods to quantify the direct ELISA of these aging biomarkers.Methods. Telomere lengths of 50 healthy persons are measured with real-time PCR in blood cells. Plasma samples from all subjects are analyzed using direct ELISA.Results. From 25 years old person to 78 years, the telomere length becomes shorter during aging. In blood plasma, the expression levels of CRAMP increases during human aging. There is the reverse correspondence between the telomere length and the plasma CRAMP level. We also find that the fresh plasma, the frozen plasma which thawed less than 3 times, and the plasma kept in the room temperature less than 3 hours are better for the ELISA analyze of CRAMP in the plasma.Conclusion. This CRAMP ELISA could become a powerful tool for investigating the relationship between human aging and telomere length shortening.
Vehicle Vibration Analysis in Changeable Speeds Solved by Pseudoexcitation Method
Authors: Guo Li-Xin , Zhang Li-Ping
Published: November 2010

The vehicle driving comfort has become one of the important factors of vehicle quality and receives increasing attention. In this paper, the mechanical and mathematical models of the half‐car, five degrees of freedom (DOF) of a vehicle were established, as well as the pseudoexcitation model of road conditions for the front wheel and the rear wheel. By the pseudoexcitation method, the equations of transient response and power spectrum density were established. After numerical simulation to vehicle vibration response of changeable driving, the results show that the pseudoexcitation method is more convenient than the traditional method and effectively solves the smoothness computation problems of vehicles while the pseudoexcitation method is used to analyze vehicle vibration under nonstationary random vibration environments.
Friend Turns Foe: Transformation of Anti-Inflammatory HDL to Proinflammatory HDL during Acute-Phase Response
Authors: G Hima Bindu , Rao Veena S. , Kakkar Vijay V.
Published: November 2010

High-density lipoprotein (HDL) is a major carrier of cholesterol in the blood. Unlike other lipoproteins, physiological functions of HDL influence the cardiovascular system in favorable ways except when HDL is modified pathologically. The cardioprotective mechanism of HDL is mainly based on reverse cholesterol transport, but there has been an emerging interest in the anti-inflammatory and antioxidant roles of HDL. These latter activities of HDL are compromised in many pathological states associated with inflammation. Further, abnormal HDL can become proinflammatory contributing to oxidative damage. In this paper, we discuss the functional heterogeneity of HDL, how alterations in these particles in inflammatory states result in loss of both antioxidant activity and reverse cholesterol transport in relation to atherosclerosis, and the need for assays to predict its functionality.
FPGA Implementation of a Pipelined Gaussian Calculation for HMM-Based Large Vocabulary Speech Recognition
Authors: Veitch Richard , Aubert Louis-Marie , Woods Roger , Fischaber Scott
Published: November 2010

A scalable large vocabulary, speaker independent speech recognition system is being developed using Hidden Markov Models (HMMs) for acoustic modeling and a Weighted Finite State Transducer (WFST) to compile sentence, word, and phoneme models. The system comprises a software backend search and an FPGA-based Gaussian calculation which are covered here. In this paper, we present an efficient pipelined design implemented both as an embedded peripheral and as a scalable, parallel hardware accelerator. Both architectures have been implemented on an Alpha Data XRC-5T1, reconfigurable computer housing a Virtex 5 SX95T FPGA. The core has been tested and is capable of calculating a full set of Gaussian results from 3825 acoustic models in 9.03 ms which coupled with a backend search of 5000 words has provided an accuracy of over 80%. Parallel implementations have been designed with up to 32 cores and have been successfully implemented with a clock frequency of 133 MHz.
Tamoxifen, Flaxseed, and the Lignan Enterolactone Increase Stroma- and Cancer Cell–Derived IL-1Ra and Decrease Tumor Angiogenesis in Estrogen-Dependent Breast Cancer
Authors: Lindahl Gabriel , Saarinen Niina , Abrahamsson Annelie , Dabrosin Charlotta
Published: November 2010

The proinflammatory cytokines IL-1α and IL-1β promote tumor angiogenesis that might be counteracted by the IL-1 receptor antagonist (IL-1Ra), anakinra, a clinically approved agent. A diet with high amounts of phytoestrogens, such as flaxseed (Flax), genistein (GEN), and the mammalian lignan enterolactone (ENL), may affect breast cancer progression in a similar fashion as the antiestrogen tamoxifen. Both cancer cells and tumor stroma may be targets for cancer therapy. By using microdialysis in a model of human breast cancers in nude mice, we could perform species-specific analyses of released proteins in the microenvironment. We show that tumors treated with tamoxifen and fed Flax or ENL exhibited decreased in vivo release of IL-1β derived from the murine stroma and decreased microvessel density whereas dietary GEN had no effects. Cancer cell–released IL-1Ra were approximately 5 times higher than stroma-derived IL-1Ra. Tamoxifen, Flax, and ENL increased IL-1Ra levels significantly whereas GEN did not. The tumor stroma contained macrophages, which expressed the estrogen receptor. In vitro, estradiol decreased IL-1Ra released from breast cancer cells and from cultured macrophages. IL-1Ra decreased endothelial cell proliferation significantly in vitro whereas breast cancer cell proliferation was unaffected in presence of estradiol. Finally, IL-1Ra therapy of tumor-bearing mice opposed estrogen-dependent breast cancer growth and decreased angiogenesis. We conclude that the release of IL-1s both by cancer cells and the stroma, where macrophages are a key component, may offer feasible targets for antiestrogen therapy and dietary interventions against breast cancer. Cancer Res; 71(1); 51–60. ©2011 AACR.
Impact ofABCB1Allelic Variants on QTc Interval Prolongation
Authors: Sissung Tristan M. , Gardner Erin R. , Piekarz Richard L. , Howden Reuben , Chen Xiaohong , Woo Sukyung , Franke Ryan , Clark James A. , Miller-DeGraff Laura , Steinberg Seth M. , Venzon David , Liewehr David , Kleeberger Steven R. , Bates Susan E. , Price Douglas K. , Rosing Douglas R. , Cabell Christopher , Sparreboom Alex , Figg William D.
Published: November 2010

Purpose: Although the ABCB1 (P-glycoprotein) drug transporter is a constituent of several blood–tissue barriers (i.e., blood–brain and blood–nerve), its participation in a putative blood–heart barrier has been poorly explored. ABCB1 could decrease the intracardiac concentrations of drugs that cause QT prolongation and cardiotoxicity.

Experimental Design: ABCB1-related romidepsin transport kinetics were explored in LLC-PK1 cells transfected with different ABCB1 genetic variants. ABCB1 plasma and intracardiac concentrations were determined in Abcb1a/1b (−/−) mice and wild-type FVB controls. These same mice were used to evaluate romidepsin-induced heart rate-corrected QT interval (QTc) prolongation over time. Finally, a cohort of 83 individuals with available QTcB and ABCB1 genotyping data were used to compare allelic variation in ABCB1 versus QTc-prolongation phenotype.

Results: Here, we show that mice lacking the ABCB1-type P-glycoprotein have higher intracardiac concentrations of a model ABCB1 substrate, romidepsin, that correspond to changes in QT prolongation from baseline (ΔQTc) over time. Consistent with this observation, we also show that patients carrying genetic variants that could raise ABCB1 expression in the cardiac endothelium have lower ΔQTc following a single dose of romidepsin.

Conclusions: To our knowledge, this is the first evidence that Abcb1-type P-glycoprotein can limit intracardiac exposure to a drug that mediates QT prolongation and suggests that certain commonly inherited polymorphisms in ABCB1 may serve as markers for QT prolongation following the administration of ABCB1-substrate drugs. Clin Cancer Res; 17(4); 937–46. ©2010 AACR.
Nitric Oxide–Donating Acetylsalicylic Acid Induces Apoptosis in Chronic Lymphocytic Leukemia Cells and Shows Strong Antitumor Efficacy In vivo
Authors: Razavi Regina , Gehrke Iris , Gandhirajan Rajesh Kumar , Poll-Wolbeck Simon Jonas , Hallek Michael , Kreuzer Karl-Anton
Published: November 2010

Purpose: Nitric oxide–donating acetylsalicylic acid (NO-ASA) has been shown to possess an antineoplastic effect in Wnt-/β-catenin–active cancers. As chronic lymphocytic leukemia (CLL) cells exhibit aberrantly active Wnt signaling, we investigated the effect of the para-isomer of NO-ASA on CLL cell survival in vitro and in a CLL-like xenograft mouse model.

Experimental Design: Apoptosis in primary CLL cells was determined by flow cytometric annexin V–FITC (fluorescein isothiocyanate)/PI (propidium iodide) staining and immunoblotting of caspases, poly(ADP-ribose) polymerase (PARP), and antiapoptotic proteins. Interference of NO-ASA with Wnt/β-catenin signaling was analyzed through immunoblots of different pathway members. Influence of caspase activation was investigated by pretreatment with a pan-caspase inhibitor. CLL-like JVM3 cells were subcutaneously inoculated into irradiated nude mice that were treated with 100 mg of para-NO-ASA/kg of body weight p.o. (by mouth) for 21 days.

Results: para-NO-ASA induced apoptosis in CLL cells with an LC50 (lethal concentration) of 8.72 + 0.04 μmol/L, whereas healthy blood cells were not affected. Furthermore, the compound induced caspase 9, caspase 3, and PARP cleavage. In addition, cleavage of β-catenin and downregulation of β-catenin/lymphoid enhancer factor (Lef)–1 targets was observed. para-NO-ASA demonstrated strong antitumor efficacy in the xenograft mouse model with a tumor inhibtion rate of 83.4%. During therapy, no gross toxicity could be observed.

Conclusions: para-NO-ASA selectively induces apoptosis in primary CLL cells and efficiently reduces tumor growth in a CLL-like xenograft model. As NO-ASA is orally available and is generally well tolerated, para-NO-ASA might be a promising new compound for CLL therapy. Clin Cancer Res; 17(2); 286–93. ©2010 AACR.
Allergen-Induced Coexpression of bFGF and TGF-β1 by Macrophages in a Mouse Model of Airway Remodeling: bFGF Induces Macrophage TGF-β1 Expression in vitro
Authors: Yum Hye Young , Cho Jae Youn , Miller Marina , Broide David H.
Published: November 2010

Background: Basic fibroblast growth factor (bFGF) is a cytokine that is mitogenic for fibroblasts and smooth muscle and may play a role in airway remodeling in asthma. We have used a mouse model of chronic ovalbumin (OVA) allergen-induced airway remodeling to determine whether bFGF and fibroblast growth factor receptor-1 are expressed and regulated by corticosteroids in the airway, as well as to determine whether bFGF mediates expression of another proremodeling cytokine, transforming growth factor (TGF)-β1. Methods: The airway levels and localization of bFGF, FGF receptor-1 and TGF-β1 were determined by ELISA, immunohistology and image analysis in the remodeled airways of chronic OVA-challenged mice treated with either corticosteroids or diluent. In vitro cultures of bone narrow-derived macrophages were used to determine whether bFGF induced TGF-β1 expression. Results: Mice chronically challenged with OVA developed significant airway remodeling that was associated with significantly increased levels of bFGF and TGF-β1. Immunohistochemistry demonstrated significantly increased bFGF and FGF receptor-1 expression by peri- bronchial F4/80+ cells. Double-label immunofluorescence microscopy studies demonstrated that peribronchial macrophages coexpressed bFGF and TGF-β1. In vitro studies demonstrated that incubation of bone marrow-derived macrophages with bFGF induced expression of TGF-β1. Mice treated with corticosteroids and subjected to chronic OVA challenge had significantly reduced levels of bFGF, FGF receptor-1, peribronchial TGF-β1+ cells and airway remodeling. Conclusions: Overall, this study demonstrates that allergen challenge stimulates peribronchial macrophages to coexpress bFGF and TGF-β1 and that bFGF may potentiate macrophage release of TGF-β1 through autocrine and/or paracrine pathways.
A Search for Biomarkers as Diagnostic Tools for Food Allergy: A Pilot Study in Peanut-Allergic Patients
Authors: Peeters Kim A.B.M. , Lamers Robert-Jan A.N. , Penninks André H. , Knol Edward F. , Bruijnzeel-Koomen Carla A.F.M. , van Nesselrooij Joop H.J. , Knulst André C.
Published: November 2010

Background: The impact of peanut allergy is large and accidental ingestion of peanut can lead to severe reactions. Currently used diagnostic tests, such as skin prick tests (SPT) and determination of specific immunoglobulins (IgE) have, however, limited sensitivity and specificity. Therefore, new tools have to be developed to improve the accuracy of the diagnostic work-up of food-allergic patients. Comprehensive metabolite analysis may provide biomarkers for diagnosing food allergy as metabolite levels reflect actual physiological conditions. We investigated whether metabolites can be found that discriminate between peanut-allergic patients and non-peanut-allergic subjects. Such metabolites may be used for future diagnostic purposes. Methods: Plasma and saliva samples were obtained from 23 participants (12 peanut allergic and 11 peanut tolerant) prior to and after a peanut challenge and measured with 1H nuclear magnetic resonance (NMR) spectroscopy with subsequent multivariate data analysis. Results: Clear differences were observed between NMR spectra of peanut-allergic and peanut-tolerant subjects in plasma as well as saliva. Allergic patients already showed aberrant metabolite levels prior to peanut ingestion, thus before the onset of allergic reactions. Conclusion: This pilot study shows that aberrant metabolite levels as determined by NMR in combination with multivariate statistics may serve as novel biomarkers for food allergy.
Flagellin Induces the Expression of Thymic Stromal Lymphopoietin in Human Keratinocytes via Toll-Like Receptor 5
Authors: Le Tuan Anh , Takai Toshiro , Vu Anh Tuan , Kinoshita Hirokazu , Chen Xue , Ikeda Shigaku , Ogawa Hideoki , Okumura Ko
Published: November 2010

Background: Thymic stromal lymphopoietin (TSLP), highly expressed by keratinocytes in skin lesions of atopic dermatitis patients and bronchial epithelial cells in asthma, plays a key role in allergic diseases. Information on triggers for the release of TSLP in keratinocytes is still limited. Keratinocytes express Toll-like receptor (TLR) 5, the ligand for which is flagellin, the major structural protein of the flagella of Gram-negative bacteria. IL-4, IL-13 and TNF-α (Th2/TNF) are associated with allergic diseases. TGF-α, one of the ligands for the epidermal growth factor receptor, is overexpressed in keratinocytes in atopic dermatitis. We investigated the induction of TSLP expression in keratinocytes stimulated with flagellin and its modulation by the Th2/TNF cytokines and TGF-α. Methods: Primary human keratinocytes were stimulated with flagellin with or without cytokines. The TSLP released was measured by ELISA. Gene expression was analyzed by quantitative real-time PCR. Results: Stimulation of keratinocytes with flagellin induced the release of TSLP protein and upregulation of the gene expression of TSLP and other pro-inflammatory molecules. The flagellin-induced release of TSLP was enhanced by the Th2/TNF cytokines or TGF-α. Small interfering RNA-mediated knockdown of TLR5 expression suppressed the flagellin-induced TSLP gene expression. Conclusions: Flagellin induces TSLP expression in keratinocytes via TLR5 and the expression can be upregulated by a cytokine milieu with Th2/TNF or TGF-α, suggesting that exposure of barrier-defective skin to Gram-negative bacteria or environmental flagellin contributes to the initiation and/or amplification of Th2-type skin inflammation including atopic dermatitis through the induction of TSLP expression in keratinocytes.
Uromodulin Biology and Pathophysiology – An Update
Authors: Vyletal Petr , Bleyer Anthony J. , Kmoch Stanislav
Published: November 2010

Uromodulin (UMOD) is a glycoprotein expressed on the luminal surface of the apical membrane of renal tubular epithelial cells forming the thick ascending limb of Henle. Here, UMOD forms filamentous structures probably ensuring water impermeability and the countercurrent gradient. The multidomain structure, cellular topology of UMOD and clinical consequences associated with UMOD dysfunction, however, suggest that it may be involved in other biological processes such as receptor-mediated endocytosis, mechanosensation of urinary flow, Wnt-signaling, cell cycle regulation and planar cell polarity. A specific, but as yet unidentified, protease(s) releases UMOD into the urine, where it probably contributes to colloid osmotic pressure, retards passage of positively charged electrolytes, prevents urinary tract infection and modulates formation of supersaturated salts and their crystals. UMOD expression, biosynthesis and excretion are regulated in a complex manner, and dysregulation is found in a wide range of pathological conditions. It is strongly reduced or absent in cases with mutations in UMOD, renin, HNF1B and other genetic disorders causing autosomal dominant hyperuricemic nephropathy. In contrast, elevated UMOD excretion may be associated with, and thus predictive of, chronic kidney disease. UMOD analysis is therefore of importance in all conditions with renal involvement and may be useful in the proper classification of renal diseases.
Analysis of Past and Present Methods of Measuring and Estimating Body Surface Area and the Resulting Evaluation of Its Doubtful Suitability to Universal Application
Authors: Ruggieri Giancarlo , Rocca Anna Rachele
Published: November 2010

Body surface area (BSA) is probably the most used tool for indexing physiological functions. The most relevant past and present measuring methods have been examined, and an analysis of selected methods used to measure and to estimate was performed. The evaluation of methods demonstrates a relationship between past and present measuring methods, but also highlights the doubtfulness of their suitability due to the criteria used to select the population for measurements and to estimate BSA by formulae, due to the weight variance according to the same height and the lack of a numeric estimate of the body shape. The use of BSA formulae generated from a population with anthropometric measures different from those of a population where the formulae are applied, and the change in today’s BSA towards 1.73 m2 can induce risks by incorrectly estimating GFR and in planning dialysis prescriptions.
Efficacy of Rifampin-Moxifloxacin-Metronidazole Combination Therapy in Hidradenitis Suppurativa
Authors: Join-Lambert Olivier , Coignard Hélène , Jais Jean-Philippe , Guet-Revillet Hélène , Poirée Sylvain , Fraitag Sylvie , Jullien Vincent , Ribadeau-Dumas Florence , Thèze Jacques , Le Guern Anne-Sophie , Behillil Sylvie , Leflèche Anne , Berche Patrick , Consigny Paul Henri , Lortholary Olivier , Nassif Xavier , Nassif Aude
Published: November 2010

Background: Antibiotics have been shown to improve hidradenitis suppurativa (HS) patients but complete remission is rare using these treatments. Objective: To assess the efficacy and safety of a combination of oral rifampin, moxifloxacin and metronidazole in long-lasting refractory HS. Methods: We retrospectively studied 28 consecutive HS patients including 6, 10 and 12 Hurley stage 1, 2 and 3 patients, respectively. Complete remission, defined as a clearance of all inflammatory lesions including hypertrophic scars, was the main outcome criterion of the study. Results: Complete remission was obtained in 16 patients, including 6/6, 8/10 and 2/12 patients with Hurley stage 1, 2 and 3, respectively (p = 0.0004). The median duration of treatment to obtain complete remission was 2.4 (range 0.9–6.5) and 3.8 months (range 1.6–7.4) in stage 1 and 2 patients, respectively, and 6.2 and 12 months in the 2 stage 3 patients. Main adverse events of the treatments were gastrointestinal disorders (64% of patients) and vaginal candidiasis (35% of females). Reversible tendinopathy and hepatitis occurred in 4 and 1 patient, respectively. Conclusions: Complete remission of refractory HS can be obtained using broad-spectrum antibiotics and Hurley staging is a prognostic factor of response to the treatment.