bluuebunny/crossref_metadata_2025 huggingface dataset:
Rifampin Stability in 7H9 Broth and Löwenstein-Jensen Medium
Authors: Yu Xia , Jiang Guanglu , Li Huiwen , Zhao Yanlin , Zhang Hongtao , Zhao Liping , Ma Yu , Coulter Christopher , Huang Hairong
Published: December 2010

The objectives of this study were to monitor the stability of rifampin (RIF) in Löwenstein-Jensen medium (L-J medium) and 7H9 broth, which are the media commonly used for drug susceptibility testing (DST) of
Mycobacterium tuberculosis
. Rifampin degradation in stock solution, 7H9 broth, and L-J medium and during the inspissation process for L-J medium preparation was serially monitored by high-performance liquid chromatography (HPLC). L-J medium-based DST was conducted to examine the effect of L-J medium storage on the DST outcome. The RIF stock solution was stable for at least 3 months when kept at either 4°C or −20°C; RIF in 7H9 broth and L-J medium was almost 50% decayed after 1 week of storage at 37°C, and rifampin could not be detected in 7H9 or L-J medium after 3 weeks or 6 weeks of storage at 37°C. Approximately half of the drug was decomposed after 4 months of storage at 4°C for both media, and after 6 months of storage at 4°C, RIF in L-J medium was undetectable, while 38% of RIF remained in 7H9 medium. Approximately 21, 24, 29, and 35% RIF degradations were detected when the L-J medium was coagulated at 75°C, 80°C, 85°C, and 90°C, respectively. The DST outcomes when using L-J medium stored for different periods of time were consistent with the RIF concentration monitoring data. Rifampin in stock solution is stable for at least 3 months at a reduced storage temperature. Media containing RIF should be prepared strictly according to validated standard operating procedures. RIF degradation is a possible reason for false resistance categorizations of
M. tuberculosis
isolates in the clinical laboratory.
Imipenem-Resistant Nocardia cyriacigeorgica Infection in a Child with Chronic Granulomatous Disease
Authors: Hagerman Andrés , Rodríguez-Nava Verónica , Boiron Patrick , Crisinel Pierre-Alex , Posfay-Barbe Klara M.
Published: December 2010

Nocardia
spp. can lead to local or disseminated infections, especially in immunocompromised patients. Combination therapy of amikacin and imipenem is commonly used to treat severe nocardial infections. We describe a patient with imipenem-resistant
Nocardia cyriacigeorgica
, which, to our knowledge, has not been previously reported among isolates of this species.
Intracellular Assembly of Cyanophage Syn5 Proceeds through a Scaffold-Containing Procapsid
Authors: Raytcheva Desislava A. , Haase-Pettingell Cameron , Piret Jacqueline M. , King Jonathan A.
Published: December 2010

Syn5 is a marine cyanophage that is propagated on the marine photosynthetic cyanobacterial strainSynechococcussp. WH8109 under laboratory conditions. Cryoelectron images of this double-stranded DNA (dsDNA) phage reveal an icosahedral capsid with short tail appendages and a single novel hornlike structure at the vertex opposite the tail. Despite the major impact of cyanophages on life in the oceans, there is limited information on cyanophage intracellular assembly processes within their photosynthetic hosts. The one-step growth curve of Syn5 demonstrated a short cycle with an eclipse period of ∼45 min, a latent phase of ∼60 min, and a burst size of 20 to 30 particles per cell at 28°C. SDS-PAGE and Western blot analysis of cell lysates at different times after infection showed the synthesis of major virion proteins and their increase as the infection progressed. The scaffolding protein of Syn5, absent from virions, was identified in the lysates and expressed from the cloned gene. It migrated anomalously on SDS-PAGE, similar to the phage T7 scaffolding protein. Particles lacking DNA but containing the coat and scaffolding proteins were purified from Syn5-infected cells using CsCl centrifugation followed by sucrose gradient centrifugation. Electron microscopic images of the purified particles showed shells lacking condensed DNA but filled with protein density, presumably scaffolding protein. These findings suggest that the cyanophages form infectious virions through the initial assembly of scaffolding-containing procapsids, similar to the assembly pathways for the enteric dsDNA bacteriophages. Since cyanobacteria predate the enteric bacteria, this procapsid-mediated assembly pathway may have originated with the cyanophages.
Efficient Culture Adaptation of Hepatitis C Virus Recombinants with Genotype-Specific Core-NS2 by Using Previously Identified Mutations
Authors: Scheel Troels K. H. , Gottwein Judith M. , Carlsen Thomas H. R. , Li Yi-Ping , Jensen Tanja B. , Spengler Ulrich , Weis Nina , Bukh Jens
Published: December 2010

Hepatitis C virus (HCV) is an important cause of chronic liver disease, and interferon-based therapy cures only 40 to 80% of patients, depending on HCV genotype. Research was accelerated by genotype 2a (strain JFH1) infectious cell culture systems. We previously developed viable JFH1-based recombinants encoding the structural proteins (core, E1, E2), p7, and NS2 of prototype isolates of the seven major HCV genotypes; most recombinants required adaptive mutations. To enable genotype-, subtype-, and isolate-specific studies, we developed efficient core-NS2 recombinants from additional genotype 1a (HC-TN and DH6), 1b (DH1 and DH5), and 3a (DBN) isolates, using previously identified adaptive mutations. Introduction of mutations from isolates of the same subtype either led to immediate efficient virus production or accelerated culture adaptation. The DH6 and DH5 recombinants without introduced mutations did not adapt to culture. Universal adaptive effects of mutations in NS3 (Q1247L, I1312V, K1398Q, R1408W, and Q1496L) and NS5A (V2418L) were investigated for JFH1-based genotype 1 to 5 core-NS2 recombinants; several mutations conferred adaptation to H77C (1a), J4 (1b), S52 (3a), and SA13 (5a) but not to ED43 (4a). The mutations permitting robust virus production in Huh7.5 cells had no apparent effect on viral replication but allowed efficient assembly of intracellular infectious HCV for adapted novel or previously developed recombinants. In conclusion, previously identified mutations permitted development of novel HCV core-NS2 genotype recombinants. Mutations adapting several recombinants to culture were identified, but no mutations were universally adaptive across genotypes. This work provides tools for analysis of HCV genotype specificity and may promote the understanding of genotype-specific patterns in HCV disease and control.
Establishment of a Chimeric, Replication-Deficient Influenza A Virus Vector by Modulation of Splicing Efficiency
Authors: Wolschek Markus , Samm Elisabeth , Seper Helena , Sturlan Sanda , Kuznetsova Irina , Schwager Cornelia , Khassidov Alexandra , Kittel Christian , Muster Thomas , Egorov Andrej , Bergmann Michael
Published: December 2010

Segment 8 of the influenza A virus codes for two proteins (NS1 and NS2/NEP) via splicing. Here, we developed a viral vector expressing a cytokine or chemokine instead of the interferon antagonist NS1. To achieve both the desired genetic stability and high transgene expression levels, NS2/NEP mRNA splicing efficacy had to be fine-tuned by modification of splicing elements. Expression levels of secreted foreign proteins could be further enhanced by fusing the N-terminal 13 amino acids of NS1 with an IgK-derived secretion signal peptide. Thus, the first start codon was used for translation initiation of both NS2/NEP and the foreign protein.
JNK1 Is Required for Lentivirus Entry and Gene Transfer
Authors: Lee Meng-Horng , Padmashali Roshan , Andreadis Stelios T.
Published: December 2010

Although a lot of progress has been made in development of lentiviral vectors for gene therapy, the interactions of these vectors with cellular factors have not been explored adequately. Here we show that lentivirus infection phosphorylates JNK and that blocking the kinase activity of JNK decreases gene transfer in a dose-dependent manner, regardless of the viral envelope glycoprotein. Knockdown by small interfering RNA (siRNA) revealed that JNK1 but not JNK2 was required for productive gene transfer. The effect of JNK on gene transfer was not due to changes in the cell cycle, as JNK knockdown did not affect the cell cycle profile of target cells and even increased cell proliferation. In addition, confluent cell monolayers also exhibited JNK phosphorylation upon lentivirus infection and a dose-dependent decrease in gene transfer efficiency upon JNK inhibition. On the other hand, JNK activation was necessary for lentivirus internalization into the cell cytoplasm, while inhibition of JNK activity decreased virus entry without affecting binding to the cell surface. These experiments suggest that JNK is required for lentivirus entry into target cells and may have implications for gene transfer or for development of antiviral agents.
Rhesus Rotavirus Entry into a Polarized Epithelium Is Endocytosis Dependent and Involves Sequential VP4 Conformational Changes
Authors: Wolf Marie , Vo Phuoc T. , Greenberg Harry B.
Published: December 2010

Rotavirus (RV) cell entry is an incompletely understood process, involving VP4 and VP7, the viral proteins composing the outermost layer of the nonenveloped RV triple-layered icosahedral particle (TLP), encasing VP6. VP4 can exist in three conformational states: soluble, cleaved spike, and folded back. In order to better understand the events leading to RV entry, we established a detection system to image input virus by monitoring the rhesus RV (RRV) antigens VP4, VP6, and VP7 at very early times postinfection. We provide evidence that decapsidation occurs directly after cell membrane penetration. We also demonstrate that several VP4 and VP7 conformational changes take place during entry. In particular, we detected, for the first time, the generation of folded-back VP5 in the context of the initiation of infection. Folded-back VP5 appears to be limited to the entry step. We furthermore demonstrate that RRV enters the cell cytoplasm through an endocytosis pathway. The endocytosis hypothesis is supported by the colocalization of RRV antigens with the early endosome markers Rab4 and Rab5. Finally, we provide evidence that the entry process is likely dependent on the endocytic Ca
2+
concentration, as bafilomycin A1 treatment as well as an augmentation of the extracellular calcium reservoir using CaEGTA, which both lead to an elevated intraendosomal calcium concentration, resulted in the accumulation of intact virions in the actin network. Together, these findings suggest that internalization, decapsidation, and cell membrane penetration involve endocytosis, calcium-dependent uncoating, and VP4 conformational changes, including a fold-back.
Foot-and-Mouth Disease Virus Exhibits an Altered Tropism in the Presence of Specific Immunoglobulins, Enabling Productive Infection and Killing of Dendritic Cells
Authors: Robinson L. , Windsor M. , McLaughlin K. , Hope J. , Jackson T. , Charleston B.
Published: December 2010

Foot-and-mouth disease virus (FMDV) causes an acute vesicular disease of farm animals. The development of successful control strategies is limited by an incomplete understanding of the immune response to FMDV. Dendritic cells (DC) mediate the induction of immunity to pathogens, but their role in FMDV infection of cattle is uncharacterized. Bovine monocyte-derived DC (moDC) were exposed to integrin-binding and cell culture-adapted strains of FMDVin vitro. MoDC were not largely susceptible to infection by integrin-binding FMDV but were susceptible to culture-adapted virus. Binding specific antibodies to integrin-binding FMDV at neutralizing or subneutralizing IgG concentrations significantly enhanced infection via CD32 (FcγR). Monocytes also expressed CD32 but were nonsusceptible to FMDV immune complex (IC) infection, indicating a requirement for additional factors involved in cellular susceptibility. Infection of moDC by the FMDV IC was productive and associated with high levels of cell death. Infected moDC were unable to efficiently stimulate FMDV-specific CD4+memory T cells, but exposing moDC to IC containing inactivated FMDV resulted in significantly increased T cell stimulation. Thus, neutralized FMDV concurrently loses its ability to infect susceptible cells while gaining the capacity to infect immune cells. This represents a change in the tropism of FMDV that could occur after the onset of the antibody response. We propose that IC could dynamically influence the anti-FMDV immune response and that this may explain why the early immune response to FMDV has evolved toward T cell independencein vivo. Moreover, we propose that DC targeting could prove useful in the development of effective vaccines against FMDV.
A Single Amino Acid Substitution in the Capsid of Foot-and-Mouth Disease Virus Can Increase Acid Resistance
Authors: Martín-Acebes Miguel A. , Vázquez-Calvo Ángela , Rincón Verónica , Mateu Mauricio G. , Sobrino Francisco
Published: December 2010

Foot-and-mouth disease virus (FMDV) particles lose infectivity due to their disassembly at pH values slightly below neutrality. This acid-dependent disassembly process is required for viral RNA release inside endosomes. To study the molecular determinants of viral resistance to acid-induced disassembly, six FMDV variants with increased resistance to acid inactivation were isolated. Infection by these mutants was more sensitive to drugs that raise the endosomal pH (NH
4
Cl and concanamycin A) than was infection by the parental C-S8c1 virus, confirming that the increase in acid resistance is related to a lower pH requirement for productive uncoating. Amino acid replacement N17D at the N terminus of VP1 capsid protein was found in all six mutants. This single substitution was shown to be responsible for increased acid resistance when introduced into an infectious FMDV clone. The increased resistance of this mutant against acid-induced inactivation was shown to be due to its increased resistance against capsid dissociation into pentameric subunits. Interestingly, the N17D mutation was located close to but not at the interpentamer interfaces. The mutants described here extend the panel of FMDV variants exhibiting different pH sensitivities and illustrate the adaptive flexibility of viral quasispecies to pH variations.
Factors Supporting Intrathecal Humoral Responses following Viral Encephalomyelitis
Authors: Phares Timothy W. , Marques Cristina P. , Stohlman Stephen A. , Hinton David R. , Bergmann Cornelia C.
Published: December 2010

Central nervous system (CNS) infections and autoimmune inflammatory disorders are often associated with retention of antibody-secreting cells (ASC). Although beneficial or detrimental contributions of ASC to CNS diseases remain to be defined, virus-specific ASC are crucial in controlling persistent CNS infection following coronavirus-induced encephalomyelitis. This report characterizes expression kinetics of factors associated with ASC homing, differentiation, and survival in the spinal cord, the prominent site of coronavirus persistence. Infection induced a vast, gamma interferon (IFN-γ)-dependent, prolonged increase in chemokine (C-X-C motif) ligand 9 (CXCL9), CXCL10, and CXCL11 mRNA, supporting a role for chemokine (C-X-C motif) receptor 3 (CXCR3)-mediated ASC recruitment. Similarly, CD4 T cell-secreted interleukin-21, a critical regulator of both peripheral activated B cells and CD8 T cells, was sustained during viral persistence. The ASC survival factors B cell-activating factor of the tumor necrosis factor (TNF) family (BAFF) and a proliferating-inducing ligand (APRIL) were also significantly elevated in the infected CNS, albeit delayed relative to the chemokines. Unlike IFN-γ-dependent BAFF upregulation, APRIL induction was IFN-γ independent. Moreover, both APRIL and BAFF were predominantly localized to astrocytes. Last, the expression kinetics of the APRIL and BAFF receptors coincided with CNS accumulation of ASC. Therefore, the factors associated with ASC migration, differentiation, and survival are all induced during acute viral encephalomyelitis, prior to ASC accumulation in the CNS. Importantly, the CNS expression kinetics implicate rapid establishment, and subsequent maintenance, of an environment capable of supporting differentiation and survival of protective antiviral ASC, recruited as plasmablasts from lymphoid organs.
Effectiveness and cost-effectiveness of an injury prevention programme for adult male amateur soccer players: design of a cluster-randomised controlled trial: Table 1
Authors: van Beijsterveldt Anna M C , Krist Mark R , Schmikli Sandor L , Stubbe Janine H , de Wit G Ardine , Inklaar Han , van de Port Ingrid G L , Backx Frank J G
Published: December 2010

Background and aims

Approximately 16% of all sports injuries in the Netherlands are caused by outdoor soccer. A cluster-randomised controlled trial has been designed to investigate the effectiveness and cost-effectiveness of an injury prevention programme (‘The11’) for male amateur soccer players. The injury prevention programme The11, developed with the support of the World Football Association FIFA, aims to reduce the impact of intrinsic injury risk factors in soccer.

Methods

Teams playing at first-class amateur level in two districts in the Netherlands are participating in the study. Teams in the intervention group were instructed to apply The11 during each practice session throughout the 2009–10 season. All participants of the control group continued their practice sessions as usual. All soccer-related injuries and related costs for each team were systematically reported online by a member of the medical staff. Player exposure to practice sessions and matches was reported weekly by the coaches. Also the use of The11 during the season after the intervention season will be monitored.

Discussion

Our hypothesis is that integrating the The11 exercises in the warm-up for each practice session is effective in terms of injury incidence, injury severity, healthcare use, and its associated costs and/or absenteeism. Prevention of soccer injuries is expected to be beneficial to adult soccer players, soccer clubs, the Royal Dutch Football Association (KNVB), health insurance companies and society.
Factors influencing investigator-caused nest abandonment by North American dabbling ducks
Authors: Garrettson P. R. , Richkus K. D. , Rohwer F. C. , Johnson W. P.
Published: December 2010

We examined factors (species, nest age, nest initiation date, clutch size, predator activity) that may affect the probability of investigator-caused nest abandonment in North American dabbling ducks and made predictions based on parental investment theory. For all nests, the best model contained species, nest stage, nest initiation date, and the interaction of species with nest initiation date. The probability of abandonment by Mallards ( Anas platyrhynchos L., 1758) was consistently higher than that of Blue-winged Teal ( Anas discors L., 1766). In these species, abandonment probability increased with later date, whereas Gadwall ( Anas strepera L., 1758), Northern Pintails ( Anas acuta L., 1758), and Northern Shovelers ( Anas clypeata L., 1758) showed the opposite pattern. Abandonment by all species declined as nest stage increased. Early-laying (≤5 eggs) females were 7 times more likely, and late-laying females were twice as likely, to abandon nests as incubating females. During incubation, abandonment probability was 38% higher during early (≤8 days) incubation than late incubation, and for each additional egg in a completed clutch, it was 19% lower. We propose a novel, two-stage model in which dabbling duck nest abandonment is influenced predominantly by opportunities for future reproduction during laying, and expected benefits from the current reproductive event during incubation.
RETURNING TO ROOTS FOR REDUCING INVENTORY COSTS IN SMEs: A CASE OF INDIAN LOCK INDUSTRY
Authors: FAROOQUIE PARVEEN , NASIR KHAN M.
Published: December 2010

Achieving uninterrupted production, increasing profitability and productivity, protection against stock-outs, and improving customer service levels have been the major benefits of holding inventory. Maintaining inventory levels beyond a certain limit is, however, an undesirable phenomenon. While practitioners and researchers are aiming at minimum possible cost of inventory, many small business owners in India still fail to appreciate fully the true costs of carrying inventory. In this context the present study examines a case of the lock manufacturing industry in the Aligarh district of the state Uttar Pradesh. The investigations reveal that lock manufacturing units, by-and-large, are either unaware of even the fundamental techniques of inventory management or do not practice them for one reason or the other. The case attempts to illustrate how SMEs can reduce their inventory costs, and hence improve profitability, through the application of basic models of inventory management.
DETERMINANTS OF ENTREPRENEURIAL INTEREST AND RISK TOLERANCE AMONG RUSSIAN UNIVERSITY STUDENTS: EMPIRICAL STUDY
Authors: KARHUNEN PÄIVI , LEDYAEVA SVETLANA
Published: December 2010

This paper, examines the determinants of entrepreneurial interest and risk tolerance among Russian university students using the ordered logit model. The data for the paper was retrieved from a survey conducted in spring of 2008 involving 200 students from three St Petersburg-based universities. The paper produced three main findings. Firstly, the students were highly interested in pursuing entrepreneurial careers and had moderate risk tolerance. Secondly, the respondents with high entrepreneurial interest included males, students of business/economic specialties, students with family business backgrounds, students with experience of being self-employed, and those students who had favourable attitudes toward entrepreneurial behaviour, perceived social norms about entrepreneurship and higher entrepreneurial self-efficacy. Thirdly, the respondents who tended to have the lowest level of risk tolerance were females, students of technical specialties, students without entrepreneurial experience and students with lower entrepreneurial self-efficacy.
SYNTHESIS AND PHOTOLUMINESCENCE OF WATER-SOLUBLE ZnS:Mn2+/ZnS QUANTUM DOTS BY NUCLEATION DOPING STRATEGY
Authors: YU ZHANGSEN , MA XIYING
Published: December 2010

We report the synthesis of luminescent-doped core/shell quantum dots (QDs) of water-soluble manganese-doped zinc sulfide (ZnS:Mn2+/ZnS) . QDs of ZnS:Mn2+/ZnS were prepared by nucleation doping strategy, with thioglycolic acid (TGA) as stabilizer in aqueous solution. Structure and optical properties of the ZnS:Mn2+/ZnS core/shell quantum dots were characterized by X-ray diffraction and photoluminescence emission spectroscopy. The influence of the synthesis conditions on the luminescent properties of ZnS:Mn2+/ZnS QDs is discussed. Different Mn2+ concentrations, ratios of the TGA/ (Zn+Mn) and thickness of the ZnS shell were used. Results showed that the ZnS:Mn2+/ZnS QDs are water-soluble and have improved fluorescence properties. Therefore, Mn2+ -doped ZnS quantum dots could be potential candidates as fluorescent labeling agents in biology.
ON MODULI SPACES AND CW STRUCTURES ARISING FROM MORSE THEORY ON HILBERT MANIFOLDS
Authors: QIN LIZHEN
Published: December 2010

This paper proves some results on negative gradient dynamical systems of Morse functions on Hilbert manifolds. It contains the compactness of flow lines, manifold structures of certain compactified moduli spaces, orientation formulas, and CW structures of the underlying manifolds.
Proinflammatory treatment of astrocytes with lipopolysaccharide results in augmented Ca2+ signaling through increased expression of VIA phospholipase A2 (iPLA2)
Authors: Strokin Mikhail , Sergeeva Marina , Reiser Georg
Published: December 2010

Many Ca2+-regulated intracellular processes are involved in the development of neuroinflammation. However, the changes of Ca2+ signaling in the brain under inflammatory conditions were hardly studied. ATP-induced Ca2+ signaling is a central event of signal transmission in astrocytic networks. We investigated primary astrocytes after proinflammatory stimulation with lipopolysaccharide (LPS; 100 ng/ml) for 6–24 h. We reveal that Ca2+ responses to purinergic ATP stimulation are significantly increased in amplitude and duration after stimulation with LPS. We detected that increased amplitudes of Ca2+ responses to ATP in LPS-treated astrocytes can be explained by substantial increase of Ca2+ load in stores in endoplasmic reticulum. The mechanism implies enhanced Ca2+ store refilling due to the amplification of capacitative Ca2+ entry. The reason for the increased duration of Ca2+ responses in LPS-treated cells is also the amplified capacitative Ca2+ entry. Next, we established that the molecular mechanism for the LPS-induced amplification of Ca2+ responses in astrocytes is increased expression and activity of VIA phospholipase A2 (VIA iPLA2). Indeed, both gene silencing with specific small interfering RNA and pharmacological inhibition of VIA iPLA2 with S-bromoenol lactone reduced the load of the Ca2+ stores and caused a decrease in the amplitudes of Ca2+ responses in LPS-treated astrocytes to values, which were comparable with those in untreated cells. Our findings highlight a novel regulatory role of VIA iPLA2 in development of inflammation in brain. We suggest that this enzyme might be a possible target for treatment of pathologies related to brain inflammation.
Treatment with IL-27 attenuates experimental colitis through the suppression of the development of IL-17-producing T helper cells
Authors: Sasaoka Tetsumasa , Ito Masayuki , Yamashita Junji , Nakajima Kenji , Tanaka Issei , Narita Masakuni , Hara Yukio , Hada Kaori , Takahashi Munehisa , Ohno Youichi , Matsuo Takato , Kaneshiro Yoshiaki , Tanaka Hitoshi , Kaneko Kenji
Published: December 2010

Inflammatory bowel disease (IBD) represents a group of chronic inflammatory diseases characterized by inflammation and relapsing gastrointestinal disorders. Recent studies have shown that Th17 cells, which are well known as key mediators of chronic inflammation, have a pivotal role in onset and development of IBD in humans and mice, alike. In recent years, it has been reported that IL-27, which is an IL-12-related heterodimeric cytokine consisting of EBI3 and p28 subunits, act directly on naive T cells to suppress the differentiation of Th17 cells. However, effects of exogenous IL-27 on the IBD are not well elucidated. To clarify the suppressive effect of IL-27 treatment on IBD, we applied the flexible linking method to EBI3 and p28 subunits and generated a single-chain human IL-27 (scIL-27). scIL-27 inhibited xenogenic mouse Th17 cell differentiation in vitro, indicating that scIL-27 also acts in mouse immune systems. In a 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced mouse acute colitis model, subcutaneous scIL-27 treatment significantly improved the colon length, extent of necrosis, and ulceration and thickened epithelium and several pathological scores in a dose-dependent manner. scIL-27 clearly suppressed several inflammatory cytokines, including IL-17, in inflamed colon, except for anti-inflammatory cytokine IL-10. The mesenteric lymph node cells from scIL-27-treated mice also exhibited a reduced inflammatory response and, furthermore, a lower population of Th17 cells than those of PBS-treated mice. Finally, we showed the therapeutic efficacy of scIL-27 on TNBS-induced colitis even after active colitis was established. These results suggest new possible therapeutic approaches for IBD, including disorders such as Crohn's disease and ulcerative colitis.
Glycerol loss to water exceeds glycerol catabolism via glycerol kinase in freeze-resistant rainbow smelt (Osmerus mordax)
Authors: Ditlecadet Delphine , Short Connie E. , Driedzic William R.
Published: December 2010

Rainbow smelt accumulate high amounts of glycerol in winter. In smelt, there is a predictable profile of plasma glycerol levels that starts to increase in November (<5 μmol/ml), peaks in mid-February (>200 μmol/ml), and thereafter decreases to reach the initial levels in the beginning of May. The aim of this study was to investigate the respective role of the two main mechanisms that might be involved in glycerol clearance from mid-February: 1) breakdown of glycerol to glycerol-3-phosphate through the action of the glycerol kinase (GK) and 2) direct loss toward the environment. Over the entire glycerol cycle, loss to water represents a daily loss of ∼10% of the total glycerol content of fish. GK activities were very low in all tissues investigated and likely have a minor quantitative role in the glycerol cycle. These results suggest that glycerol levels are dictated by the rate of glycerol synthesis (accelerated and deactivated during the accumulation and decrease stages, respectively). Although not important in glycerol clearance, GK in liver might have an important metabolic function for other purposes, such as gluconeogenesis, as evidenced by the significant increase of activity at the end of the cycle.
Glomerular sclerosis is prevented during urinary tract obstruction due to podocyte protection
Authors: Matsusaka Taiji , Kobayashi Kazuto , Kon Valentina , Pastan Ira , Fogo Agnes B. , Ichikawa Iekuni
Published: December 2010

Urine outflow obstruction activates a variety of profibrotic factors, including the intrarenal renin-angiotensin system. However, the obstruction also nullifies the transmural hydraulic pressure difference across the glomerular capillary wall, an established inducer of glomerulosclerosis. In the present study, we investigated whether, and by what mechanism, urine outflow obstruction affects the process of progressive glomerulosclerosis. For this purpose, we tested the effect of unilateral ureteral obstruction (UUO) of 7 days duration in two distinct mouse models of glomerulosclerosis. In the human immunodeficiency virus (HIV) nephropathy model, where HIV-1 genes are selectively expressed in podocytes and develop progressive podocyte damage and glomerulosclerosis, UUO protected against sclerosis with preservation of podocytes morphologically and immunohistochemically. In contrast, the nonobstructed contralateral kidneys of these mice, as well as sham-operated HIV-1 mouse kidneys, developed severe podocyte injury and glomerulosclerosis. The protection against glomerulosclerosis imparted by ureteral obstruction was also documented in the NEP25 model of podocyte injury, in which a single injection of immunotoxin, LMB2, triggers selective podocyte injury followed by glomerulosclerosis, both of which were protected by UUO. Notably, intervention with an angiotensin II type 1 receptor antagonist provided only a partial protective effect in each of the models. These results demonstrate that urine outflow obstruction protects the glomerulus from progressive sclerosis. The results further reveal that this protection occurs at a very early stage of the pathologic process, namely, damage of podocytes.
Dynamical Properties of a Delay Prey‐Predator Model with Disease in the Prey Species Only
Authors: Shi Xiangyun , Zhou Xueyong , Song Xinyu
Published: December 2010

A three‐dimensional ecoepidemiological model with delay is considered. We first investigate the existence and stability of the equilibria. We then study the effect of the time delay on the stability of the positive equilibrium. The existence of a Hopf bifurcation at the positive equilibrium is obtained through the study of an
exponential polynomial equation with delay‐dependent coefficients. Numerical simulation with a hypothetical set of data has been carried out to support the analytical findings.
Role of Meconium and Hypoxia in Meconium Aspiration-Induced Lung Injury in Neonatal Rabbits
Authors: Zagariya Alex , Sierzputovska Monica , Navale Shan , Vidyasagar Dharmapuri
Published: December 2010

Background. We previously showed that meconium cuases lung cell death by apoptosis and inflammatory cytokine expression. Whether this is due to meconium exposure itself, or meconium related hypoxia remains unclear.Objectives. To elucidate the effects of meconium, saline, milk, hypoxia and hyperoxia induced lung injury.Design/Methods. We studied 5 groups of rabbit pups: (I) normal saline; (II) Milk; (III) 10% solution of meconium; (IV) only to 15 minutes of hypoxia (10% O2), and (V) 5 minutes of hypoxia (95% O2). After exposure lung lavage cells were used for apoptotic cell count and cytokine expression.In vitroresponse of human A 549 epithelial cells to meconium-and milk exposure was also studied.Results. There was no difference in cell death between saline and milk groups. However, meconium caused a significant cell loss compared to saline and milk—Inflammatory cytokines increased significantly in meconium group compared to saline or milk group. Although hypoxic and hyperoxic lungs showed increased inflammatory reaction compared to saline-treated lungs, this injury was not significant compared to meconium group. Studies with A549 cells also showed similar results.Conclusions. We conclude that lung cell injury in meconium aspiration is maily from meconium itself.
Use of Nebulized Amphotericin B in the Treatment of Allergic Bronchopulmonary Aspergillosis in Cystic Fibrosis
Authors: Proesmans M. , Vermeulen F. , Vreys M. , De Boeck K.
Published: December 2010

Background. Systemic steroids and adjunctive antifungal therapy are the cornerstone in treating allergic bronchopulmonary aspergillosis (ABPA) in the context of CF.Aim. Evaluate the use of inhaled amphotericin B (iAMB) as antifungal agent in this context.Methods. Report of 7 CF patients with recurrent or difficult to treat ABPA and failure to taper systemic corticosteroids treated with AMB deoxycholate (AMB-d) (Fungizone 25 mg 3× a week) or AMB lipid complex (ABLC) (Abelcet 50 mg twice weekly). Successful therapy was defined as steroid withdrawal without ABPA relapse within 12 months.Results. Therapy was successful in 6 of 7 patients treated with iAMB. In 5/6, lung function improved. The patient with treatment failure has concomitant MAC lung infection.Conclusion. Inhaled AMB may be an alternative to commonly used adjunctive antifungal therapy in the treatment of ABPA. More data are needed on safety and efficacy.
Molecular Characterization of Putative Chordoma Cell Lines
Authors: Brüderlein Silke , Sommer Joshua B. , Meltzer Paul S. , Li Sufeng , Osada Takuya , Ng David , Möller Peter , Alcorta David A. , Kelley Michael J.
Published: December 2010

Immortal tumor cell lines are an important model system for cancer research, however, misidentification and cross-contamination of cell lines are a common problem. Seven chordoma cell lines are reported in the literature, but none has been characterized in detail. We analyzed gene expression patterns and genomic copy number variations in five putative chordoma cell lines (U-CH1, CCL3, CCL4, GB60, and CM319). We also created a new chordoma cell line, U-CH2, and provided genotypes for cell lines for identity confirmation. Our analyses revealed that CCL3, CCL4, and GB60 are not chordoma cell lines, and that CM319 is a cancer cell line possibly derived from chordoma, but lacking expression of key chordoma biomarkers. U-CH1 and U-CH2 both have gene expression profiles, copy number aberrations, and morphology consistent with chordoma tumors. These cell lines also harbor genetic changes, such as loss of p16, MTAP, or PTEN, that make them potentially useful models for studying mechanisms of chordoma pathogenesis and for evaluating targeted therapies.
Novel Organotin(IV)‐Schiff Base Complexes: Synthesis, Characterization, Antimicrobial Activity, and DNA Interaction Studies
Authors: Prasad K. Shiva , Kumar L. Shiva , Prasad Melvin , Revanasiddappa Hosakere D.
Published: December 2010

Four organotin(IV) complexes with 2‐(2‐hydroxybenzylideneamino)isoindoline‐1,3‐dione (L1), and 4‐(4‐hydroxy‐3‐methoxybenzylideneamino‐N‐(pyrimidin‐2‐yl)benzenesulfonamide (L2) were synthesized and well characterized by analytical and spectral studies. The synthesized compounds were tested for antimicrobial activity by disc diffusion method. The DNA binding of the complexes 1 and 3 with CT‐DNA has been performed with absorption spectroscopy, which showed that both the complexes are avid binders of CT‐DNA. Also the nuclease activity of complexes 1 and 3 with plasmid DNA (pUC19) was studied using agarose gel electrophoresis. The complex 1 can act as effective DNA cleaving agent when compared to complex 3 resulting in the nicked form of DNA under physiological conditions. The gel was run both in the absence and presence of the oxidizing agent.
Application of a Chairside Anaerobic Culture Test for Endodontic Treatment
Authors: Yoneda Masahiro , Kita Seidai , Suzuki Nao , Macedo Sonia M. , Iha Kosaku , Hirofuji Takao
Published: December 2010

Periapical lesions are caused by bacterial infections. The fundamental objective of endodontic treatment is to eliminate bacteria present in the root canal system because they play an important role in the development and maintenance of periapical lesions. Therefore, confirming the absence of bacteria before filling root canals is important. Anaerobic culture tests have been used in many endodontic cases, and they have brought about good treatment outcomes. These tests, however, require specific apparatuses and bacteriological techniques. Here, we report a chairside anaerobic culture test that does not require any specialized apparatuses or techniques. We also report two endodontic cases in which this simple test was used. Both patients were diagnosed with chronic purulent periapical lesions. After confirming the absence of bacteria in the root canals, they were filled with gutta-percha points. At followup, the radiolucencies showed recovery, although longterm observation is under way. From these results, the authors conclude that this simple chairside anaerobic culture test is effective for evaluating periapical lesion treatment procedures.
Impact of Orlistat-Induced Weight Loss on Diastolic Function and Heart Rate Variability in Severely Obese Subjects with Diabetes
Authors: Martin Julie , Paquette Carmen , Marceau Simon , Hould Frédéric-Simon , Lebel Stéfane , Simard Serge , Dumesnil Jean-Gaston , Poirier Paul
Published: December 2010

Objective. Determine the impact of Orlistat-induced weight loss on metabolic profile and cardiovascular function in severely obese patients with type 2 diabetes.Methods. Twenty-nine patients were randomized either to a nonplacebo control group or to a treatment group with Orlistat thrice a day. Metabolic profile, anthropometric parameters, heart rate variability indices, and echocardiographic variables were measured before and after a 12-week treatment period.Results. Treatment with Orlistat induced a modest but significant weight loss compared to controls (3.7 ± 3.0 versus 0.5 ± 2.2 kg, resp.;P=.003). There was significant decrease in fasting glycemia (7.9 ± 3.0 versus 6.7 ± 2.2 mmol/L;P=.03) and significant improvements in left ventricular diastolic function (P=.03) and in the sympathovagal balance (LF/HF ratio) (P=.04) in the Orlistat group.Conclusion. These results suggest that a modest weight loss improves fasting glycemia, left ventricular diastolic function, and sympathovagal balance in severely obese patients with type 2 diabetes.
An Update on the Genetics of Usher Syndrome
Authors: Millán José M. , Aller Elena , Jaijo Teresa , Blanco-Kelly Fiona , Gimenez-Pardo Ascensión , Ayuso Carmen
Published: December 2010

Usher syndrome (USH) is an autosomal recessive disease characterized by hearing loss, retinitis pigmentosa (RP), and, in some cases, vestibular dysfunction. It is clinically and genetically heterogeneous and is the most common cause underlying deafness and blindness of genetic origin. Clinically, USH is divided into three types. Usher type I (USH1) is the most severe form and is characterized by severe to profound congenital deafness, vestibular areflexia, and prepubertal onset of progressive RP. Type II (USH2) displays moderate to severe hearing loss, absence of vestibular dysfunction, and later onset of retinal degeneration. Type III (USH3) shows progressive postlingual hearing loss, variable onset of RP, and variable vestibular response. To date, five USH1 genes have been identified:MYO7A(USH1B),CDH23(USH1D),PCDH15(USH1F),USH1C(USH1C), andUSH1G(USH1G). Three genes are involved in USH2, namely,USH2A(USH2A),GPR98(USH2C), andDFNB31(USH2D). USH3 is rare except in certain populations, and the gene responsible for this type isUSH3A.
Construction, Characterization, and Preliminary BAC‐End Sequence Analysis of a Bacterial Artificial Chromosome Library of the Tea Plant (Camellia sinensis)
Authors: Lin Jinke , Kudrna Dave , Wing Rod A.
Published: December 2010

We describe the construction and characterization of a publicly available BAC library for the tea plant, Camellia sinensis. Using modified methods, the library was constructed with the aim of developing public molecular resources to advance tea plant genomics research. The library consists of a total of 401,280 clones with an average insert size of 135 kb, providing an approximate coverage of 13.5 haploid genome equivalents. No empty vector clones were observed in a random sampling of 576 BAC clones. Further analysis of 182 BAC‐end sequences from randomly selected clones revealed a GC content of 40.35% and low chloroplast and mitochondrial contamination. Repetitive sequence analyses indicated that LTR retrotransposons were the most predominant sequence class (86.93%–87.24%), followed by DNA retrotransposons (11.16%–11.69%). Additionally, we found 25 simple sequence repeats (SSRs) that could potentially be used as genetic markers.
Current Concepts: Mouse Models of Sjögren′s Syndrome
Authors: Lavoie Tegan N. , Lee Byung Ha , Nguyen Cuong Q.
Published: December 2010

Sjögren′s syndrome (SjS) is a complex chronic autoimmune disease of unknown etiology which primarily targets the exocrine glands, resulting in eventual loss of secretory function. The disease can present as either primary SjS or secondary SjS, the latter of which occurs concomitantly with another autoimmune disease such as rheumatoid arthritis, systemic lupus erythematosus, scleroderma, or primary biliary cirrhosis. Current advancements in therapeutic prevention and treatment for SjS are impeded by lack of understanding in the pathophysiological and clinical progression of the disease. Development of appropriate mouse models for both primary and secondary SjS is needed in order to advance knowledge of this disease. This paper details important features, advantages, and pitfalls of current animal models of SjS, including spontaneous, transgenic, knockout, immunization, and transplantation chimera mouse models, and emphasizes the need for a better model in representing the human SjS phenotype.
Iterative Methods for Variational Inequalities over the Intersection of the Fixed Points Set of a Nonexpansive Semigroup in Banach Spaces
Authors: Mohamadi Issa
Published: December 2010

This paper presents a framework of iterative methods for finding specific common fixed points of a nonexpansive self-mappings semigroup in a Banach space. We prove, with appropriate conditions, the strong convergence to the solution of some variational inequalities.
Correlating 18FDG PET-CT with SPET-CT Lymphoscintigraphy Optimizes the Surgical Management of the Axilla in Patients with T2 or T3 Breast Cancer.
Authors: Soler C. , Bouteille C. , Steiner A. , Geissler B. , Grémillet E.
Published: December 2010

Purpose: the aim of this study was to assess the impact of 18 FDG PET-CT (FDG-PET) and sentinel lymph node (SLN) localization by lymphoSPET-CT for the surgical management of the axilla in patients with T2 or T3 breast cancer.Patients and methods: 69 patients with T2 or T3 breast cancer (20 to 40 mm) were enrolled. Both FDG-PET and lymphoSPET-CT were performed for all 69 patients. FDG-PET was performed first to analyze breast and axillary area. Two to 4 days after, SLN SPET-CT was performed (4 to 12 hours before SLN biopsy and breast surgery).When a focal accumulation of FDG was noted and related to one or more possible axillary metastases, it was compared with SLN as identified by SPECT-CT lymphoscintigraphy. Concordance and discordance of node localization were noted.Results: a focal axillary FDG uptake was found in 30 patients (43,4%). For these 30 patients, preoperative lymphoSPET-CT revealed SLN in 26. All the 26 SLNs were in the same CT location as the focal FDG uptake. SLN biopsy procedure was done in these 26 patients and revealed node metastases in 16 cases (61, 5%). Then, axillary lymph node dissection (ALND) was performed when SLN was metastatic.For the 4 patients with focal axillary FDG uptake but no SLN detected by lymphoSPET-CT, ALND was performed and revealed axillary metastases in all cases (4/4).For the 39 patients (56.6%) for whom no focal FDG uptake around axillary region was found, lymphoSPET-CT always identified SLN which was when biopsied and followed by ALND only when metastatic. 3/39 (7,5%) SLN were metastatic, 36/39 (92, 5%) SLN were healthy.So correlating 18FDG PET-CT with lymphoSPET-CT seems to be very important in cases of T2 and T3 breast cancer:1- When a focal FDG uptake is found and no SLN is found by lymphoSPET-CT, ALND is indicated.2- When a focal FDG uptake is found and is anatomically correlated with SLN as identified by lymphoSPET-CT, SLN biopsy procedure seems to be allowed and safe because node biopsy is related certainly with PET suspect node3- In case of no focal FDG uptake in axillary area, SLN biopsy procedure is probably as safe as for T1 patients.Conclusion: if these preliminary results were to be confirmed, correlating 18FDG PET-CT with SPET-CT lymphoscintigraphy will allow avoiding systematic ALND in patients with T2 or T3 breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 1024.