bluuebunny/crossref_metadata_2025 huggingface dataset:
190
nm
excimer laser drilling of glass slices: Dependence of drilling rate and via hole shape on the diameter of the via hole
Authors: Chen K. H. , Wu Wenhsing , Chu Byung Hwan , Lo C. F. , Lin Jenshan , Wang Y. L. , Chang C. Y. , Pearton S. J. , Ren F.
Published: November 2009

The authors have investigated the effect of the via hole diameter on the laser drilling rate of glass as well as the shape of the drilled via holes. An Ar–F2 based 193nm UV excimer laser was used in this study. The via holes with a diameter of 120μm showed a 7.5°–9° angled, tapered side wall, and the drilling rate was relatively constant at around 17μm∕s. For the smaller via holes with diameters ranging from 30–80μm, significantly different results were obtained due to laser reflection from the tapered side wall of the via hole leading to the drilling rate being slightly increased and via hole becoming conical in shape. For the smallest via holes with an entrance diameter of 10μm, the resulting hole showed a very high aspect ratio, with a funnel shaped via hole and a significantly reduced drilling rate.
Frequent Release of Low Amounts of Herpes Simplex Virus from Neurons: Results of a Mathematical Model
Authors: Schiffer Joshua T. , Abu-Raddad Laith , Mark Karen E. , Zhu Jia , Selke Stacy , Magaret Amalia , Wald Anna , Corey Lawrence
Published: November 2009

A stochastic mathematical model suggests that very low numbers of herpes simplex virus 2 particles are released frequently into the genital tract in infected individuals.
Transcriptomic Response ofListeria monocytogenesto Iron Limitation andfurMutation
Authors: Ledala Nagender , Sengupta Mrittika , Muthaiyan Arunachalam , Wilkinson Brian J. , Jayaswal R. K.
Published: November 2009

Iron is required by almost all bacteria, but concentrations above physiological levels are toxic. In bacteria, intracellular iron is regulated mostly by the ferric uptake regulator, Fur, or a similar functional protein. Iron limitation results in the regulation of a number of genes, especially those involved in iron uptake. A subset of these genes is the Fur regulon under the control of Fur. In the present study, we have identified Fur- and iron-regulated genes inListeria monocytogenesby DNA microarray analysis using afurmutant and its isogenic parent. To identify genes regulated exclusively in response to iron limitation, the whole-genome transcriptional responses to the iron limitation of afurmutant and its isogenic parent were compared. Fur-regulated genes were identified by comparing the transcriptional profile of the parent with the transcriptional profile of the isogenicfurmutant. Our studies have identified genes regulated exclusively in response to iron and those that are negatively regulated by Fur. We have identified at least 14 genes that were negatively regulated directly by Fur. Under iron-limited conditions, these genes were upregulated, while the expression offurwas found to be downregulated. To further investigate the regulation offurin response to iron, an ectopicfurpromoter-lacZtranscriptional fusion strain was constructed, and its isogenicfurandperRmutant derivatives were generated inL. monocytogenes10403S. Analysis of the iron limitation of theperRmutant indicated that the regulation of genes under the negative control of Fur was significantly inhibited. Our results indicate that Fur and PerR proteins negatively regulatefurand that under iron-limited conditions, PerR is required for the negative regulation of genes controlled by Fur.
Improved Risk Analysis by Dual Direct Detection of Total and Infectious
Cryptosporidium
Oocysts on Cell Culture in Combination with Immunofluorescence Assay
Authors: Lalancette Cindy , Di Giovanni George D. , Prévost Michèle
Published: November 2009

The inactivation of
Cryptosporidium
oocysts is a main driver in the selection of water treatment disinfection strategies, and microbial risk analysis provides a sound basis for optimizing water treatment processes. U.S. Environmental Protection Agency method 1622/23 provides an estimate of the total oocyst count; however, it cannot be used directly for risk assessment, as it does not determine the fraction of infectious oocysts. Improved assessment of the risk for designated sources or in treated water requires evaluation of the total number of oocysts and an estimate of their infectivity. We developed a dual direct detection method using differential immunofluorescent staining that allows detection of both oocysts and cell culture infection foci for each sample. Using
Cryptosporidium parvum
oocysts, various pH levels, proteases, and gastroenteric compounds and substrates were assessed to determine their abilities to enhance the number of infection foci. The results showed that the key trigger for oocyst stimulation was acidification. Addition of a low concentration of
d
-glucose (50 mM) to the infection media increased rates of infectivity, while a higher dose (300 mM) was inhibitory. The total number of oocysts in each sample was determined by counting the oocysts remaining on a cell monolayer and the oocysts recovered from cell monolayer washes during processing using a simple filtration technique. With the dual direct detection on cell culture with immunofluorescence assay method, it is now possible to determine the numbers of total and infectious oocysts for a given sample in a single analysis. Direct percentages of infectivity are then calculated, which allows more accurate assessments of risk.
Cotranslocation of Methyl Parathion Hydrolase to the Periplasm and of Organophosphorus Hydrolase to the Cell Surface of
Escherichia coli
by the Tat Pathway and Ice Nucleation Protein Display System
Authors: Yang Chao , Freudl Roland , Qiao Chuanling , Mulchandani Ashok
Published: November 2009

A genetically engineered
Escherichia coli
strain coexpressing organophosphorus hydrolase (OPH) and methyl parathion hydrolase (MPH) was constructed for the first time by cotransforming two compatible plasmids. Since these two enzymes have different substrate specificities, the coexpression strain showed a broader substrate range than strains expressing either one of the hydrolases. To reduce the mass transport limitation of organophosphates (OPs) across the cell membrane, MPH and OPH were simultaneously translocated to the periplasm and cell surface of
E. coli
, respectively, by employing the twin-arginine translocation (Tat) pathway and ice nucleation protein (INP) display system. The resulting recombinant strain showed sixfold-higher whole-cell activity than the control strain expressing cytosolic OP hydrolases. The correct localization of MPH and OPH was demonstrated by cell fractionation, immunoblotting, and enzyme activity assays. No growth inhibition was observed for the recombinant
E. coli
strain, and suspended cultures retained almost 100% of the activity over a period of 2 weeks. Owing to its high level of activity and superior stability, the recombinant
E. coli
strain could be employed as a whole-cell biocatalyst for detoxification of OPs. This strategy of utilizing dual translocation pathways should open up new avenues for cotranslocating multiple functional moieties to different extracytosolic compartments of a bacterial cell.
Iron Sources Used by the Nonpathogenic Lactic Acid Bacterium
Lactobacillus sakei
as Revealed by Electron Energy Loss Spectroscopy and Secondary-Ion Mass Spectrometry
Authors: Duhutrel Philippe , Bordat Christian , Wu Ting-Di , Zagorec Monique , Guerquin-Kern Jean-Luc , Champomier-Vergès Marie-Christine
Published: November 2009

Lactobacillus sakei
is a lactic acid bacterium naturally found on meat. Although it is generally acknowledged that lactic acid bacteria are rare species in the microbial world which do not have iron requirements, the genome sequence of
L. sakei
23K has revealed quite complete genetic equipment dedicated to transport and use of this metal. Here, we aimed to investigate which iron sources could be used by this species as well as their role in the bacterium's physiology. Therefore, we developed a microscopy approach based on electron energy loss spectroscopy (EELS) analysis and nano-scale secondary-ion mass spectrometry (SIMS) in order to analyze the iron content of
L. sakei
cells. This revealed that
L. sakei
can use iron sources found in its natural ecosystem, myoglobin, hemoglobin, hematin, and transferrin, to ensure long-term survival during stationary phase. This study reveals that analytical image methods (EELS and SIMS) are powerful complementary tools for investigation of metal utilization by bacteria.
Analysis of Lipid Export in Hydrocarbonoclastic Bacteria of the GenusAlcanivorax: Identification of Lipid Export-Negative Mutants ofAlcanivorax borkumensisSK2 andAlcanivorax jadensisT9
Authors: Manilla-Pérez Efraín , Reers Christina , Baumgart Meike , Hetzler Stephan , Reichelt Rudolf , Malkus Ursula , Kalscheuer Rainer , Wältermann Marc , Steinbüchel Alexander
Published: November 2009

Triacylglycerols (TAGs), wax esters (WEs), and polyhydroxyalkanoates (PHAs) are the major hydrophobic compounds synthesized in bacteria and deposited as cytoplasmic inclusion bodies when cells are cultivated under imbalanced growth conditions. The intracellular occurrence of these compounds causes high costs for downstream processing.Alcanivoraxspecies are able to produce extracellular lipids when the cells are cultivated on hexadecane or pyruvate as the sole carbon source. In this study, we developed a screening procedure to isolate lipid export-negative transposon-induced mutants of bacteria of the genusAlcanivoraxfor identification of genes required for lipid export by employing the dyes Nile red and Solvent Blue 38. Three transposon-induced mutants ofA. jadensisand seven ofA. borkumensisimpaired in lipid secretion were isolated. All isolated mutants were still capable of synthesizing and accumulating these lipids intracellularly and exhibited no growth defect. In theA. jadensismutants, the transposon insertions were mapped in genes annotated as encoding a putative DNA repair system specific for alkylated DNA (Aj17), a magnesium transporter (Aj7), and a transposase (Aj5). In theA. borkumensismutants, the insertions were mapped in genes encoding different proteins involved in various transport processes, like genes encoding (i) a heavy metal resistance (CZCA2) in mutant ABO_6/39, (ii) a multidrug efflux (MATE efflux) protein in mutant ABO_25/21, (iii) an alginate lyase (AlgL) in mutants ABO_10/30 and ABO_19/48, (iv) a sodium-dicarboxylate symporter family protein (GltP) in mutant ABO_27/29, (v) an alginate transporter (AlgE) in mutant ABO_26/1, or (vi) a two-component system protein in mutant ABO_27/56. Site-directedMATE,algE, andalgLgene disruption mutants, which were constructed in addition, were also unable to export neutral lipids and confirmed the phenotype of the transposon-induced mutants. The putative localization of the different gene products and their possible roles in lipid excretion are discussed. Beside this, the composition of the intra- and extracellular lipids in the wild types and mutants were analyzed in detail.
Thioredoxin System from
Deinococcus radiodurans
Authors: Obiero Josiah , Pittet Vanessa , Bonderoff Sara A. , Sanders David A. R.
Published: November 2009

This paper describes the cloning, purification, and characterization of thioredoxin (Trx) and thioredoxin reductase (TrxR) and the structure determination of TrxR from the ionizing radiation-tolerant bacterium
Deinococcus radiodurans
strain R1. The genes from
D. radiodurans
encoding Trx and TrxR were amplified by PCR, inserted into a pET expression vector, and overexpressed in
Escherichia coli.
The overexpressed proteins were purified by metal affinity chromatography, and their activity was demonstrated using well-established assays of insulin precipitation (for Trx), 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB) reduction, and insulin reduction (for TrxR). In addition, the crystal structure of oxidized TrxR was determined at 1.9-Å resolution. The overall structure was found to be very similar to that of
E. coli
TrxR and homodimeric with both NADPH- and flavin adenine dinucleotide (FAD)-binding domains containing variants of the canonical nucleotide binding fold, the Rossmann fold. The

K
m

(5.7 μM) of
D. radiodurans
TrxR for
D. radiodurans
Trx was determined and is about twofold higher than that of the
E. coli
thioredoxin system. However,
D. radiodurans
TrxR has a much lower affinity for
E. coli
Trx (

K
m

, 44.4 μM). Subtle differences in the surface charge and shape of the Trx binding site on TrxR may account for the differences in recognition. Because it has been suggested that TrxR from
D. radiodurans
may have dual cofactor specificity (can utilize both NADH and NADPH),
D. radiodurans
TrxR was tested for its ability to utilize NADH as well. Our results show that
D. radiodurans
TrxR can utilize only NADPH for activity.
GyrA Interacts with MarR To Reduce Repression of themarRABOperon inEscherichia coli
Authors: Domain Francis , Levy Stuart B.
Published: November 2009

Bacterial two-hybrid studies of randomly clonedEscherichia coliDNA identified a physical interaction between GyrA, subunit A of gyrase, and MarR, a repressor of themarRABoperon. GyrA-His immobilized on Ni-nitrilotriacetic acid (NiNTA) resin bound MarR, while MarR alone did not bind. GyrA interfered with MarR binding tomarO, as detected by electrophoretic mobility assays. In a strain bearing themarRABoperon and amarO-lacZreporter, overexpression of GyrA increased LacZ activity, indicating decreased repression ofmarO-lacZby MarR. These results were confirmed by an increased survival of cells treated with quinolones and other antibiotics when GyrA was overexpressed. This work, like a previous study examining TktA (12), shows that unrelated proteins can regulate MarR activity. The findings reveal an unexpected regulatory function of GyrA in antibiotic resistance.
Genetic Variants of Human Parvovirus B19 in South Africa: Cocirculation of Three Genotypes and Identification of a Novel Subtype of Genotype 1
Authors: Corcoran Craig , Hardie Diana , Yeats Jane , Smuts Heidi
Published: November 2009

Parvovirus B19 comprises three distinct genotypes (1, 2, and 3). The distribution of B19 genotypes has not before been examined in South Africa. Two hundred thirty-nine laboratory samples submitted to a diagnostic virology laboratory for parvovirus DNA detection were analyzed retrospectively. Of the 53 PCR-positive samples investigated, 40 (75.4%) were identified as genotype 1 by genotype-specific PCR or consensus NS1 PCR and sequencing and 3 (5.7%) as genotype 2 and 10 (18.9%) as genotype 3 by analysis of NS1 sequences. Furthermore, phylogenetic analysis identified two genotype 1 sequences which were distinct from the previously described genotypes 1A and 1B. Interestingly, a genotype 2 virus was detected in the serum of an 11-year-old child, providing evidence for its recent circulation. This is the first study to demonstrate the concurrent circulation of all three genotypes of B19 in South Africa and the provisional identification of a novel subtype of genotype 1. The implications of parvovirus B19 variation are discussed.
First Report of
Gordonibacter pamelaeae
Bacteremia
Authors: Woo Patrick C. Y. , Teng Jade L. L. , Lam Kay K. M. , Tse Cindy W. S. , Leung Kit-Wah , Leung Anthony W. S. , Lau Susanna K. P. , Yuen Kwok-Yung
Published: November 2009

We report the first case of
Gordonibacter pamelaeae
bacteremia, identified by phenotypic tests and 16S rRNA sequencing in a patient with disseminated rectosigmoid carcinoma and responsive to amoxicillin-clavulanate. The bacterium was a nonsporulating, anaerobic, Gram-positive, nonmotile, coccobacillus that was catalase, arginine dihydrolase, and arginine acrylamidase positive. The gastrointestinal tract is probably its reservoir.
Transmission of Methicillin-Resistant
Staphylococcus aureus
to Household Contacts
Authors: Mollema F. P. N. , Richardus J. H. , Behrendt M. , Vaessen N. , Lodder W. , Hendriks W. , Verbrugh H. A. , Vos M. C.
Published: November 2009

The frequency of and risk factors for methicillin-resistant
Staphylococcus aureus
(MRSA) transmission from a MRSA index person to household contacts were assessed in this prospective study. Between January 2005 and December 2007, 62 newly diagnosed MRSA index persons (46 patients and 16 health care workers) and their 160 household contacts were included in the study analysis. Transmission of MRSA from an index person to household contacts occurred in nearly half of the cases (47%;
n
= 29). These 29 index persons together had 84 household contacts, of which two-thirds (67%;
n
= 56) became MRSA positive. Prolonged exposure time to MRSA at home was a significant risk factor for MRSA transmission to household contacts. In addition, MRSA colonization at least in the throat, younger age, and eczema in index persons were significantly associated with MRSA transmission; the presence of wounds was negatively associated with MRSA transmission. Furthermore, an increased number of household contacts and being the partner of a MRSA index person were household-related risk factors for MRSA acquisition from the index person. No predominant pulsed-field gel electrophoresis (PFGE) type was observed to be transmitted more frequently than other PFGE types. To date, screening household contacts and providing MRSA eradication therapy to those found positive simultaneously with the index person is not included in the “search-and-destroy” policy. We suggest including both in MRSA prevention guidelines, as this may reduce further spread of MRSA.
Theiler's Murine Encephalomyelitis Virus L* Amino Acid Position 93 Is Important for Virus Persistence and Virus-Induced Demyelination
Authors: Stavrou Spyridon , Baida Gleb , Viktorova Ekaterina , Ghadge Ghanashyam , Agol Vadim I. , Roos Raymond P.
Published: November 2009

The DA strain and other members of the TO subgroup of Theiler's murine encephalomyelitis virus (TMEV) induce a persistent central nervous system infection associated with an inflammatory white matter demyelinating disease. TO subgroup strains synthesize an 18-kDa protein, L*, out of frame with the polyprotein from an initiation codon 13 nucleotides downstream from the polyprotein's AUG codon. We previously generated a mutant virus from our infectious DA full-length clone that has a change of the L* AUG codon to ACG (with no change in the polyprotein's amino acid sequence). Studies of this mutant virus showed that L* was key to the TO subgroup phenotype because the mutant had a decreased ability to persist and demyelinate. This work was initially called into question because a similar mutant derived from a different full-length DA infectious clone persisted and demyelinated similarly to wild-type DA virus (O. van Eyll and T. Michiels, J. Virol. 74:9071-9077, 2000). We now report that (i) the sequence of the L* coding region differs in the two infectious clones, resulting in a Ser or Leu as the predicted amino acid at position 93 of L* (with no change in the polyprotein's amino acid sequence), (ii) the difference in this amino acid is key to the phenotypic differences between the two mutants, and (iii) the change in amino acid 93 may affect L* phosphorylation. It is of interest that this amino acid only appears critical in determining the virus phenotype when L* is present in a significantly reduced amount (i.e., following translation from an ACG initiating codon).
Multimerization of Hepatitis Delta Antigen Is a Critical Determinant of RNA Binding Specificity
Authors: Lin Brian C. , Defenbaugh Dawn A. , Casey John L.
Published: November 2009

Hepatitis delta virus (HDV) RNA forms an unbranched rod structure that is associated with hepatitis delta antigen (HDAg) in cells replicating HDV. Previous in vitro binding experiments using bacterially expressed HDAg showed that the formation of a minimal ribonucleoprotein complex requires an HDV unbranched rod RNA of at least about 300 nucleotides (nt) and suggested that HDAg binds the RNA as a multimer of fixed size. The present study specifically examines the role of HDAg multimerization in the formation of the HDV ribonucleoprotein complex (RNP). Disruption of HDAg multimerization by site-directed mutagenesis was found to profoundly alter the nature of RNP formation. Mutant HDAg proteins defective for multimerization exhibited neither the 300-nt RNA size requirement for binding nor specificity for the unbranched rod structure. The results unambiguously demonstrate that HDAg binds HDV RNA as a multimer and that the HDAg multimer is formed prior to binding the RNA. RNP formation was found to be temperature dependent, which is consistent with conformational changes occurring on binding. Finally, analysis of RNPs constructed with unbranched rod RNAs successively longer than the minimum length indicated that multimeric binding is not limited to the first HDAg bound and that a minimum RNA length of between 604 and 714 nt is required for binding of a second multimer. The results confirm the previous proposal that HDAg binds as a large multimer and demonstrate that the multimer is a critical determinant of the structure of the HDV RNP.
Retrograde Axon Transport of Herpes Simplex Virus and Pseudorabies Virus: a Live-Cell Comparative Analysis
Authors: Antinone Sarah Elizabeth , Smith Gregory Allan
Published: November 2009

Upon entry, neuroinvasive herpesviruses traffic from axon terminals to the nuclei of neurons resident in peripheral ganglia, where the viral DNA is deposited. A detailed analysis of herpes simplex virus type 1 (HSV-1) transport dynamics in axons following entry is currently lacking. Here, time lapse fluorescence microscopy was used to compare the postentry viral transport of two neurotropic herpesviruses: HSV-1 and pseudorabies virus (PRV). HSV-1 capsid transport dynamics were indistinguishable from those of PRV and did not differ in neurons of human, mouse, or avian origin. Simultaneous tracking of capsids and tegument proteins demonstrated that the composition of actively transporting HSV-1 is remarkably similar to that of PRV. This quantitative assessment of HSV-1 axon transport following entry demonstrates that HSV-1 and PRV share a conserved mechanism for postentry retrograde transport in axons and provides the foundation for further studies of the retrograde transport process.
The pH of Activation of the Hemagglutinin Protein Regulates H5N1 Influenza Virus Pathogenicity and Transmissibility in Ducks
Authors: Reed Mark L. , Bridges Olga A. , Seiler Patrick , Kim Jeong-Ki , Yen Hui-Ling , Salomon Rachelle , Govorkova Elena A. , Webster Robert G. , Russell Charles J.
Published: November 2009

While the molecular mechanism of membrane fusion by the influenza virus hemagglutinin (HA) protein has been studied extensively
in vitro
, the role of acid-dependent HA protein activation in virus replication, pathogenesis, and transmission
in vivo
has not been characterized. To investigate the biological significance of the pH of activation of the HA protein, we compared the properties of four recombinant viruses with altered HA protein acid stability to those of wild-type influenza virus A/chicken/Vietnam/C58/04 (H5N1)
in vitro
and in mallards. Membrane fusion by wild-type virus was activated at pH 5.9. Wild-type virus had a calculated environmental persistence of 62 days and caused extensive morbidity, mortality, shedding, and transmission in mallards. An N114K mutation that increased the pH of HA activation by 0.5 unit resulted in decreased replication, genetic stability, and environmental stability. Changes of +0.4 and −0.5 unit in the pH of activation by Y23H and K58I mutations, respectively, reduced weight loss, mortality, shedding, and transmission in mallards. An H24Q mutation that decreased the pH of activation by 0.3 unit resulted in weight loss, mortality, clinical symptoms, and shedding similar to those of the wild type. However, the HA-H24
1
Q virus was shed more extensively into drinking water and persisted longer in the environment. The pH of activation of the H5 HA protein plays a key role in the propagation of H5N1 influenza viruses in ducks and may be a novel molecular factor in the ecology of influenza viruses. The data also demonstrate that H5N1 neuraminidase activity increases the pH of activation of the HA protein
in vitro
.
Hepatoprotective Effect ofShidagonglaoon Acute Liver Injury Induced by Carbon Tetrachloride
Authors: Chao Jung , Lee Meng-Shiou , Amagaya Sakae , Liao Jiunn-Wang , Wu Jin-Bin , Ho Li-Kang , Peng Wen-Huang
Published: November 2009

This study investigates the hepatoprotective activity of ethanol extract from Shidagonglao roots (SDGLEtOH). The hepatoprotective effect of SDGLEtOH(20, 100 and 500 mg/kg) was analyzed on carbon tetrachloride ( CCl4)-induced acute liver injury. Rats pretreated orally with SDGLEtOH(100 and 500 mg/kg) and silymarin (200 mg/kg) for 3 consecutive days prior to the administration of a single dose of 50% CCl4(0.10 ml/100 g of bw, ip) significantly prevented the increases in the activities of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in CCl4-treated rats. Histological analysis also showed that SDGLEtOH(100 and 500 mg/kg) and silymarin reduced the incidence of liver lesions including vacuole formation, neutrophil infiltration and necrosis of hepatocytes induced by CCl4in rats. Moreover, the SDGLEtOH(100 and 500 mg/kg) increased the activities of anti-oxidative enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GRd) and decreased malondialdehyde (MDA) level in liver, as compared to those in the CCl4-treated group. Furthermore, SDGLEtOH(100 and 500 mg/kg) and silymarin attenuated the increased levels of tumor necrosis factor-α (TNF-α) in serum and nitric oxide ( NO ) in liver as compared to the CCl4-treated group. The hepatoprotective mechanisms of SDGLEtOHare likely related to inhibition of TNF-α, MDA and NO productions via increasing the activities of antioxidant enzymes (SOD, GPx and GRd). These experimental results suggest that SDGLEtOHcan attenuate CCl4-induced acute liver injury in rats.
Rhizomes of Gastrodia elata BL Possess Antidepressant-Like Effect via Monoamine Modulation in Subchronic Animal Model
Authors: Chen Pei-Ju , Hsieh Ching-Liang , Su Kuan-Pin , Hou Yu-Chi , Chiang Hsiu-Mei , Sheen Lee-Yan
Published: November 2009

Depression is one of the most common psychiatric disorders that affects over 10% of individuals. However there is no desirable therapeutic approach nowadays. Thus, looking for an alternative treatment is important. The objectives of this study are to investigate the antidepressant effects of the rhizomes of Gastradia elata BL, a traditional Chinese medicine, in the animal model forced-swimming test (FST), and to analyze the monoamine content in the rats' brain regions in order to better understand the possible mechanisms of the water extract of the rhizomes of Gastrodia elata BL (WGE). Subchronic administration of 0.5 or 1.0 mg/kg bw WGE by gavage for 21 consecutive days significantly reduced the immobility duration of rats in the test session of FST as compared to the control group (p < 0.05), with the equivalent effects as the antidepressant drug fluoxetine. The concentration of serotonin (5-HT) in the frontal cortex and dopamine (DA) in striatum were significantly increased after WGE administration (p < 0.05). The ratio of 5-HIAA/5-HT and (DOPAC + HVA)/DA, i.e. the turnover of 5-HT and DA respectively, were also significantly lowered in WGE groups (p < 0.05). In summary, the present study showed that the WGE possesses antidepressant-like effect on rats in the animal model FST, and the possible mechanism may be via regulating both the serotonergic and dopaminergic systems.
Anti-Proliferative Efficacy of Icariin on HepG2 Hepatoma and Its Possible Mechanism of Action
Authors: Yang Jin-Xia , Fichtner Iduna , Becker Monika , Lemm Margit , Wang Xue-Mei
Published: November 2009

The aim of the present work was to explore the anti-hepatoma effects of icariin both in vitro and in vivo and to elucidate its potential mechanism of action. The MTT assay was applied to test the anti-proliferative effects of icariin in vitro. HepG2 bearing NMRI nu/nu mice were used to test the anticancer effects of icariin in vivo. Immunohistochemical assay and flow cytometry assay (FACS) were applied to detect the possible mechanisms of action of icariin. MTT assay illustrated that icariin inhibited the proliferation of HepG2 cells in a concentration dependent manner; meanwhile, icariin inhibited the tumor growth in HepG2 bearing NMRI nu/nu mice. The tumor weight was inhibited by 55.6% and tumor volume was inhibited by 47.2%. Icariin did not influence the spleen and body weights or blood parameters. Immunohistochemical analysis indicated that the expressions of both CD31 and Ki67 in the icariin treated group were significantly lower than those in the control group (p < 0.01). FACS assay showed that icariin dramatically decreased the percentage of CD4+ and CD8+ cells in bone marrow and CD19+ cells in blood on day 8. On day 17, the percentage of CD8+ cells in blood was lower than those in the control group. CD4/CD8 ratio in icariin group was significantly elevated in bone marrow on day 17. Icariin showed anticancer efficacy both in vitro and in vivo. The possible mechanism of action could be related to its anti-angiogenesis and anti-proliferative effects in tumors.
Anti-Inflammatory Effect of Resveratrol by Inhibition of IL-8 Production in LPS-Induced THP-1 Cells
Authors: Oh You-Chang , Kang Ok-Hwa , Choi Jang-Gi , Chae Hee-Sung , Lee Young-Seob , Brice Obiang-Obounou , Jung Hyun Ju , Hong Seung-Heon , Lee Young-Mi , Kwon Dong-Yeul
Published: November 2009

Resveratrol is a polyphenol compound and prominent anti-inflammatory agent found in plants, including the fruits of Morus alba. However, the therapeutic mechanisms of resveratrol remain largely unclear. To gain insight into the biological effects of resveratrol, we examined its influence on LPS-induced IL-8 production in the human monocytic cell line, THP-1. In inflammatory diseases, IL-8 plays a central role in the initiation and maintenance of inflammatory response. In the present study, IL-8 production was measured by ELISA and RT-PCR, while MAPK activation, IκBα degradation, nuclear factor (NF)-κB activation and cyclooxygenase (COX)-2 expression were determined by Western blot analysis. Resveratrol inhibited LPS-induced IL-8 production in a dose-dependent manner. Furthermore, resveratrol inhibited extracellular signal-regulated kinase (ERK) and p38 MAPK phosphorylation, IκBα degradation, NF-κB activation and cyclooxygenase (COX)-2 expression, which suggest that resveratrol inhibits IL-8 secretion by blocking MAPK phosphorylation and NF-κB activation. Taken together, these findings may help elucidate the mechanism by which resveratrol modulates THP-1 cell activation under inflammatory conditions.
Concept Maps as a Tool for Supporting Knowledge Management in Collaborative Research Projects
Authors: Loss Leandro
Published: November 2009

The successful writing and management of collaborative research projects requires a common understanding of the project's objectives by all partners involved. It also requires a clear vision of the project inputs and outputs. This paper presents two case-studies where concept maps were used as a knowledge management tool in order to support both project life-cycle management and project proposal writing. Concept maps were drawn in both cases in order to describe the main activities executed in the project work packages (WPs) and to show how the WPs were linked to each other.
Empirical Validation of Knowledge Packages as Facilitators for Knowledge Transfer
Authors: Ardimento Pasquale , Baldassarre Maria Teresa , Cimitile Marta , Visaggio Giuseppe
Published: November 2009

Transfer of research results in production systems requires, among others, that knowledge be explicit and understandable by stakeholders. Such transfer is demanding, as so many researchers have been studying alternative ways to classic approaches such as books and papers that favour knowledge acquisition on behalf of users. In this context, we propose the concept of Knowledge Experience Package (KEP) with a specific structure as an alternative. The KEP contains both the conceptual model(s) of the research results which make up the innovation, including all the necessary documentation ranging from papers or book chapters; and the experience collected in acquiring it in business processes, appropriately structured. The structure allows the identification of the knowledge chunk(s) that the developer, who is acquiring the knowledge, needs in order to simplify the acquisition process. The experience is needed to point out the scenarios that the user will most likely face and therefore refer to. Both structure and experience are important factors for the innovation transferability and efficacy. Furthermore, we have carried out an experiment which compared the efficacy of this instrument with the classic ones, along with the comprehensibility of the information enclosed in a KEP rather than in a set of Papers. The experiment has pointed out that knowledge packages are more effective than traditional ones for knowledge transfer.
THE ROLE OF COLLABORATION, MARKET AND INTELLECTUAL PROPERTY RIGHTS AWARENESS IN UNIVERSITY TECHNOLOGY COMMERCIALIZATION
Authors: GOVINDARAJU V. G. R. CHANDRAN , GHAPAR FARHA ABDOL , PANDIYAN VEERA
Published: November 2009

University technology commercialization has become an important topic of discussion in an era where the results of university research may create opportunities for new processes and products. Despite the government's allocation of research grants to universities in developing countries such as Malaysia, the level of technology commercialization is still low. While numerous factors contribute to the poor commercialization rate, the internal dynamics are one of the major driving factors. It is in this context that we analyze the influence of awareness of market requirements and intellectual property issues and collaboration on technology commercialization. Taking the researchers as the unit of analysis, while controlling for experience and other individual characteristics, we found that researchers' awareness of market requirements, intellectual property rights and university-industry collaborations play an important role in determining the success of university technology commercialization.
INVOLVED PARTIES' CONTENTMENT WITH ENVIRONMENTAL IMPACT ASSESSMENT SYSTEM IN ESTONIA
Authors: PÕDER TÕNIS , LUKKI TIIT
Published: November 2009

Besides other approaches, interviewing main actors (decision-makers, consultants, developers) can provide valuable information about their subjective attitude as well as indicate probable weak areas and help in formulation of strategy for further research and EIA system development. This paper considers results of the survey conducted in Estonia in early 2008 as a part of national EIA system analysis. The survey covered main actors' contentment with different aspects such as EIA legislation, public participation, EIA outcomes, experts' qualification and impartiality, etc. As evidenced by what they have pointed out, the EIA system leaves much to be desired. All actors seemed to be worried about the low effectiveness of EIA. Deficiencies in public participation were also brought forward. Decision-makers trusted consultants' qualifications, but many of them felt that consultants were biased. However, the obtained results did not indicate any correlation between expert bias and contentment with EIA outcomes, perceived by decision-makers.
ASYMPTOTIC COHOMOLOGY OF CIRCULAR UNITS
Authors: BELLIARD JEAN-ROBERT
Published: November 2009

Let F be a number field, abelian over ℚ, and fix a prime p ≠ 2. Consider the cyclotomic ℤp-extension F∞/F and denote Fn the nth finite subfield and Cn its group of circular units as defined by Sinnott. Then the Galois groups Gm,n = Gal (Fm/Fn) act naturally on the Cm 's (for any m ≥ n ≫ 0). We compute the Tate cohomology groups [Formula: see text] for i = -1,0 without assuming anything else neither on F nor on p.
ARITHMETICAL RESULTS ON CERTAIN q-SERIES, II
Authors: BUNDSCHUH PETER , VÄÄNÄNEN KEIJO
Published: November 2009

As in Part I, entire transcendental solutions of certain mth order linear q-difference equations are investigated arithmetically, where now the polynomial coefficients are much more general. The purpose of this paper is to produce again lower bounds for the dimension of the K-vector space generated by 1 and the values of these solutions at m successive powers of q, where K is the rational or an imaginary quadratic field. A new feature in the proof is to use simultaneously positive and negative powers of q as interpolation points leading to an extra parameter in the main result extending its applicability.
Extensive mononuclear infiltration and myogenesis characterize recovery of dysferlin-null skeletal muscle from contraction-induced injuries
Authors: Roche Joseph A. , Lovering Richard M. , Roche Renuka , Ru Lisa W. , Reed Patrick W. , Bloch Robert J.
Published: November 2009

We studied the response of dysferlin-null and control skeletal muscle to large- and small-strain injuries to the ankle dorsiflexors in mice. We measured contractile torque and counted fibers retaining 10-kDa fluorescein dextran, necrotic fibers, macrophages, and fibers with central nuclei and expressing developmental myosin heavy chain to assess contractile function, membrane resealing, necrosis, inflammation, and myogenesis. We also studied recovery after blunting myogenesis with X-irradiation. We report that dysferlin-null myofibers retain 10-kDa dextran for 3 days after large-strain injury but are lost thereafter, following necrosis and inflammation. Recovery of dysferlin-null muscle requires myogenesis, which delays the return of contractile function compared with controls, which recover from large-strain injury by repairing damaged myofibers without significant inflammation, necrosis, or myogenesis. Recovery of control and dysferlin-null muscles from small-strain injury involved inflammation and necrosis followed by myogenesis, all of which were more pronounced in the dysferlin-null muscles, which recovered more slowly. Both control and dysferlin-null muscles also retained 10-kDa dextran for 3 days after small-strain injury. We conclude that dysferlin-null myofibers can survive contraction-induced injury for at least 3 days but are subsequently eliminated by necrosis and inflammation. Myogenesis to replace lost fibers does not appear to be significantly compromised in dysferlin-null mice.
Stimulatory effects of arachidonic acid on myosin ATPase activity and contraction of smooth muscle via myosin motor domain
Authors: Katayama Takeshi , Watanabe Masaru , Tanaka Hideyuki , Hino Mizuki , Miyakawa Takuya , Ohki Takashi , Ye Li-Hong , Xie Ce , Yoshiyama Shinji , Nakamura Akio , Ishikawa Ryoki , Tanokura Masaru , Oiwa Kazuhiro , Kohama Kazuhiro
Published: November 2009

We have been searching for a mechanism to induce smooth muscle contraction that is not associated with phosphorylation of the regulatory light chain (RLC) of smooth muscle myosin (Nakamura A, Xie C, Zhang Y, Gao Y, Wang HH, Ye LH, Kishi H, Okagaki T, Yoshiyama S, Hayakawa K, Ishikawa R, Kohama K. Biochem Biophys Res Commun 369: 135–143, 2008). In this article, we report that arachidonic acid (AA) stimulates ATPase activity of unphosphorylated smooth muscle myosin with maximal stimulation (Rmax) of 6.84 ± 0.51 relative to stimulation by the vehicle and with a half-maximal effective concentration (EC50) of 50.3 ± 4.2 μM. In the presence of actin, Rmaxwas 1.72 ± 0.08 and EC50was 26.3 ± 2.3 μM. Our experiments with eicosanoids consisting of the AA cascade suggested that they neither stimulated nor inhibited the activity. Under conditions that did not allow RLC to be phosphorylated, AA stimulated contraction of smooth muscle tissue with an Rmaxof 1.45 ± 0.07 and an EC50of 27.0 ± 4.4 μM. In addition to the ATPase activities of the myosin, AA stimulated those of heavy meromyosin, subfragment 1 (S1), S1 from which the RLC was removed, and a recombinant heavy chain consisting of the myosin head. The stimulatory effects of AA on these preparations were about twofold. The site of AA action was indicated to be the step-releasing inorganic phosphate (Pi) from the reaction intermediate of the myosin-ADP-Picomplex. The enhancement of Pirelease by AA was supported by computer simulation indicating that AA docked in the actin-binding cleft of the myosin motor domain. The stimulatory effect of AA was detectable with both unphosphorylated myosin and the myosin in which RLC was fully phosphorylated. The AA effect on both myosin forms was suggested to cause excess contraction such as vasospasm.
Changes in left ventricular function and remodeling after myocardial infarction in hypothyroid rats
Authors: Chen Yue-Feng , Redetzke Rebecca A. , Said Suleman , Beyer April J. , Gerdes A. Martin
Published: November 2009

It has been shown that hypothyroidism may lead to delayed wound healing after experimental myocardial infarction (MI) in rats and increased infarct size in dogs. However, the long-term effect of hypothyroidism on left ventricular (LV) remodeling after MI has not been determined. Adult female Sprague-Dawley rats with and without surgical thyroidectomy (TX) were used in the study. Four weeks after TX, MI or sham MI was performed on TX and non-TX rats. Rats from all groups were examined 4 wk later. Four weeks after TX, hypothyroid-induced LV dysfunction was confirmed by echocardiography. In terminal experiments 4 wk after MI, TX sham-MI rats showed smaller hearts and impaired LV function compared with non-TX sham-MI controls. TX + MI rats showed smaller hearts with bigger infarct areas, higher LV end-diastolic pressures, and greater impairment of relaxation (−dP/d t) compared with non-TX MI rats. Relative changes after MI between TX and non-TX rats for most other hemodynamic and echocardiographic indexes were similar. These results suggest that preexisting hypothyroidism exaggerates post-MI remodeling and worsens LV function, particularly diastolic function.
Mitochondria depletion abolishes agonist-induced Ca2+plateau in airway smooth muscle cells: potential role of H2O2
Authors: Chen Taoxiang , Zhu Liping , Wang Tao , Ye Hong , Huang Kewu , Hu Qinghua
Published: November 2009

The mechanisms by which mitochondria regulate the sustained phase of agonist-induced responses in cytosolic Ca2+concentration as an independent organelle in whole is not clear. By exposing to ethidium bromide and supplying pyruvate and uridine, we established mitochondrial DNA (mtDNA)-depleted rat airway smooth muscle cells (RASMCs) with maintained cellular energy. Upon an exposure to 2 μM histamine, [Ca2+]iin control RASMCs increased to a peak followed by a plateau above baseline, whereas [Ca2+]iin mtDNA-depleted RASMCs jumped to a peak and then declined to baseline without any plateau. mtDNA depletion apparently attenuated intracellular reactive oxygen species generation induced by histamine. By coexposure to 2 μM histamine and 0.1 μM exogenous H2O2, which did not affect [Ca2+]iby itself, the above difference in [Ca2+]ikinetics in mtDNA-depleted RASMCs was reversed. Intracellular H2O2decomposition abolishes histamine-induced sustained elevation in [Ca2+]iin RASMCs. Thus, mitochondria regulate agonist-induced sustained [Ca2+]ielevation by a H2O2-dependent mechanism.
Constitutively active endothelial Notch4 causes lung arteriovenous shunts in mice
Authors: Miniati Doug , Jelin Eric B. , Ng Jennifer , Wu Jianfeng , Carlson Timothy R. , Wu Xiaoqing , Looney Mark R. , Wang Rong A.
Published: November 2009

Lung arteriovenous (AV) shunts or malformations cause significant morbidity and mortality in several distinct clinical syndromes. For most patients with lung AV shunts, there is still no optimal treatment. The underlying molecular and cellular etiology for lung AV shunts remains elusive, and currently described animal models have insufficiently addressed this problem. Using a tetracycline-repressible system, we expressed constitutively active Notch4 (Notch4*) specifically in the endothelium of adult mice. More than 90% of mice developed lung hemorrhages and respiratory insufficiency and died by 6–7 wk after gene expression began. Vascular casting and fluorescent microsphere analysis showed evidence of lung AV shunts in affected mice. Cessation of Notch4* expression reversed these pathophysiological effects. Assessment of the vascular morphology revealed enlarged, tortuous vessels in the lungs that resembled arteriovenous malformations. By using whole lung organ culture, we demonstrated the effects of constitutively active Notch4 on the lung vasculature to be a primary lung phenomenon. Together, our results indicate the importance of Notch signaling in maintaining the lung vasculature and offer a new, reliable model with which to study the pathobiology of lung arteriovenous shunts and malformations.
Effect of dietary probiotics on clownfish: a molecular approach to define how lactic acid bacteria modulate development in a marine fish
Authors: Avella Matteo A , Olivotto Ike , Silvi Stefania , Place Allen R. , Carnevali Oliana
Published: November 2009

We set out to determine whether probiotic addition would improve larval development in the false percula clownfish Amphiprion ocellaris and to determine what molecular responses could be observed in the larvae following probiotic exposure. We supplied the probiotic bacterial strain Lactobacillus rhamnosus IMC 501 to clownfish larvae from the first day posthatch simultaneously by live prey and with addition to rearing water ( group 2) and exclusively by live prey ( group 3). We observed twofold higher body weight in both clownfish larvae and juveniles when probiotics were supplied via live prey and added to the rearing water. In addition, development was accelerated with metamorphosis occurring 3 days earlier in fingerlings treated with probiotic. Alteration in molecular biomarkers supported the faster growth observation. There was significantly increased gene expression of factors involved in growth and development (insulin-like growth factors I and II, myostatin, peroxisome proliferator-activated receptors α and β, vitamin D receptor α, and retinoic acid receptor γ) when probiotics were delivered via live prey and added to the rearing water. Moreover, probiotic treatment lessened the severity of the general stress response as exhibited by lower levels of glucocorticoid receptor and 70-kDa heat shock protein gene expression. Furthermore, an improvement of skeletal head development was observed, with a 10–20% reduction in deformities for juveniles treated with probiotic. All data suggest a potent effect on development resulting from the administration of lactic acid bacteria to larval clownfish, and this study provides a preliminary molecular entry path into the investigation of mechanisms responsible for probiotic enhancement in fish development.