bluuebunny/crossref_metadata_2025_split huggingface dataset:
High-Throughput PCR Assays To Monitor Wolbachia Infection in the Dengue Mosquito (Aedes aegypti) and Drosophila simulans
Authors: Lee Siu F. , White Vanessa L. , Weeks Andrew R. , Hoffmann Ary A. , Endersby Nancy M. 
Published: April 2012

ABSTRACT

We have developed and validated two new fluorescence-based PCR assays to detect the
Wolbachia w
Mel strain in
Aedes aegypti
and the
w
Ri and
w
Au strains in
Drosophila simulans
. The new assays are accurate, informative, and cost-efficient for large-scale
Wolbachia
screening.
Squid-Derived Chitin Oligosaccharides Are a Chemotactic Signal during Colonization by Vibrio fischeri
Authors: Mandel Mark J. , Schaefer Amy L. , Brennan Caitlin A. , Heath-Heckman Elizabeth A. C. , DeLoney-Marino Cindy R. , McFall-Ngai Margaret J. , Ruby Edward G. 
Published: April 2012

ABSTRACT

Chitin, a polymer of
N
-acetylglucosamine (GlcNAc), is noted as the second most abundant biopolymer in nature. Chitin serves many functions for marine bacteria in the family
Vibrionaceae
(“vibrios”), in some instances providing a physical attachment site, inducing natural genetic competence, and serving as an attractant for chemotaxis. The marine luminous bacterium
Vibrio fischeri
is the specific symbiont in the light-emitting organ of the Hawaiian bobtail squid,
Euprymna scolopes
. The bacterium provides the squid with luminescence that the animal uses in an antipredatory defense, while the squid supports the symbiont's nutritional requirements.
V. fischeri
cells are harvested from seawater during each host generation, and
V. fischeri
is the only species that can complete this process in nature. Furthermore, chitin is located in squid hemocytes and plays a nutritional role in the symbiosis. We demonstrate here that chitin oligosaccharides produced by the squid host serve as a chemotactic signal for colonizing bacteria.
V. fischeri
uses the gradient of host chitin to enter the squid light organ duct and colonize the animal. We provide evidence that chitin serves a novel function in an animal-bacterial mutualism, as an animal-produced bacterium-attracting synomone.
Heterologous Expression in Pichia pastoris and Characterization of an Endogenous Thermostable and High-Glucose-Tolerant β-Glucosidase from the Termite Nasutitermes takasagoensis
Authors: Uchima Cristiane Akemi , Tokuda Gaku , Watanabe Hirofumi , Kitamoto Katsuhiko , Arioka Manabu 
Published: April 2012

ABSTRACT

Termites are well-known cellulose decomposers and can give researchers insights into how to utilize lignocellulosic biomass in the actual scenario of energy consumption. In this work, an endogenous β-glucosidase from the midgut of the higher termite
Nasutitermes takasagoensis
was purified to homogeneity by Ni
2+
affinity chromatography and its properties were characterized. This β-glucosidase (G1mgNtBG1), which belongs to glycoside hydrolase family 1, is a homotrimer in its native form, with a molecular mass of 169.5 kDa, as demonstrated by gel filtration chromatography. The enzyme displayed maximum activity at pH 5.5 and had broad substrate specificities toward several saccharides, including cellobiose. G1mgNtBG1 showed a relatively high temperature optimum of 65°C and one of the highest levels of glucose tolerance among several β-glucosidases already characterized, with a

K
i

of 600 mM glucose. To examine the applicability of G1mgNtBG1 in biomass conversion, we compared the thermostability and glucose tolerance of G1mgNtBG1 with those of Novozym 188. We found that G1mgNtBG1 was more thermostable after 5 h of incubation at 60°C and more resistant to glucose inhibition than Novozym 188. Furthermore, our result suggests that G1mgNtBG1 acts synergistically with Celluclast 1.5 L in releasing reducing sugars from Avicel. Thus, G1mgNtBG1 seems to be a potential candidate for use as a supplement in the hydrolysis of biomass.
Characterization of anO-Demethylase of Desulfitobacterium hafniense DCB-2
Authors: Studenik Sandra , Vogel Michaela , Diekert Gabriele 
Published: April 2012

ABSTRACTBesides acetogenic bacteria, onlyDesulfitobacteriumhas been described to utilize and cleave phenyl methyl ethers under anoxic conditions; however, no ether-cleavingO-demethylases from the latter organisms have been identified and investigated so far. In this study, genes of an operon encodingO-demethylase components ofDesulfitobacterium hafniensestrain DCB-2 were cloned and heterologously expressed inEscherichia coli. Methyltransferases I and II were characterized. Methyltransferase I mediated the ether cleavage and the transfer of the methyl group to the superreduced corrinoid of a corrinoid protein.Desulfitobacteriummethyltransferase I had 66% identity (80% similarity) to that of the vanillate-demethylating methyltransferase I (OdmB) ofAcetobacterium dehalogenans. The substrate spectrum was also similar to that of the latter enzyme; however,Desulfitobacteriummethyltransferase I showed a higher level of activity for guaiacol and used methyl chloride as a substrate. Methyltransferase II catalyzed the transfer of the methyl group from the methylated corrinoid protein to tetrahydrofolate. It also showed a high identity (∼70%) to methyltransferases II ofA. dehalogenans. The corrinoid protein was produced inE. colias cofactor-free apoprotein that could be reconstituted with hydroxocobalamin or methylcobalamin to function in the methyltransferase I and II assays. Six COG3894 proteins, which were assumed to function as activating enzymes mediating the reduction of the corrinoid protein after an inadvertent oxidation of the corrinoid cofactor, were studied with respect to their abilities to reduce the recombinant reconstituted corrinoid protein. Of these six proteins, only one was found to catalyze the reduction of the corrinoid protein.
Effects of the SpoVT Regulatory Protein on the Germination and Germination Protein Levels of Spores of Bacillus subtilis
Authors: Ramirez-Peralta Arturo , Stewart Kerry-Ann V. , Thomas Stacy K. , Setlow Barbara , Chen Zhan , Li Yong-qing , Setlow Peter 
Published: April 2012

ABSTRACT

Bacillus subtilis
isolates lacking the SpoVT protein, which regulates gene expression in developing forespores, gave spores that released their dipicolinic acid (DPA) via germinant receptor (GR)-dependent germination more rapidly than wild-type spores. Non-GR-dependent germination via dodecylamine was more rapid with
spoVT
spores, but germination via Ca-DPA was slower. The effects of a
spoVT
mutation on spore germination were seen with spores made in rich and poor media, and levels of SpoVT-LacZ were elevated 2-fold in poor-medium spores; however, elevated SpoVT levels were not the only cause of the slower GR-dependent germination of poor-medium spores. The
spoVT
spores had ≥5-fold higher GerA GR levels, ∼2-fold elevated GerB GR levels, wild-type levels of a GerK GR subunit and the GerD protein required for normal GR-dependent germination, ∼2.5-fold lower levels of the SpoVAD protein involved in DPA release in spore germination, and 30% lower levels of DNA protective α/β-type small, acid-soluble spore proteins. With one exception, the effects on protein levels in
spoVT
spores are consistent with the effects of SpoVT on forespore transcription. The
spoVT
spores were also more sensitive to UV radiation and outgrew slowly. While
spoVT
spores' elevated GR levels were consistent with their more rapid GR-dependent germination, detailed analysis of the results suggested that there is another gene product crucial for GR-dependent spore germination that is upregulated in the absence of SpoVT. Overall, these results indicate that SpoVT levels during spore formation have a major impact on the germination and the resistance of the resultant spores.
Can a single question provide an accurate measure of physical activity?
Authors: Milton Karen , Clemes Stacy , Bull Fiona 
Published: April 2012

ObjectiveThe ‘single-item measure’ was developed as a short self-report tool for assessing physical activity. The aim of this study was to test the criterion validity of the single-item measure against accelerometry.DesignParticipants (n=66, 65% female, age: 39±11 years) wore an accelerometer (ActiGraph GT3X) over a 7-day period and on day 8, completed the single-item measure. The number of days of ≥30 min of accelerometer-determined moderate to vigorous intensity physical activity (MVPA) were calculated using two approaches; first by including all minutes of MVPA and second by including only MVPA accumulated in bouts of ≥10 min (counts/min ≥1952). Associations between the single-item measure and accelerometer were examined using Spearman correlations and 95% limits of agreement. Percent agreement and κ statistic were used to assess agreement between the tools in classifying participants as sufficiently/insufficiently active.ResultsCorrelations between the number of days of ≥30 min MVPA recorded by the single-item and accelerometer ranged from 0.46 to 0.57. Participants underreported their activity on the single-item measure (−1.59 days) when compared with all objectively measured MVPA, but stronger congruence was observed when compared with MVPA accumulated in bouts of ≥10 min (0.38 days). Overall agreement between the single-item and accelerometry in classifying participants as sufficiently/insufficiently active was 58% (k=0.23, 95% CI 0.05 to 0.41) when including all MVPA and 76% (k=0.39, 95% CI 0.14 to 0.64) when including activity undertaken in bouts of ≥10 min.ConclusionsThe single-item measure is a valid screening tool to determine whether respondents are sufficiently active to benefit their health.
The molecular characterisation of unusual subcutaneous spindle cell lesion of breast
Authors: Takano Elena A , Rogers Toni-Maree , Young Richard J , Rayoo Mukta , Kostos Phillip , Ferguson Ross , Campbell Ian G , Debiec-Rychter Maria , Fox Stephen B 
Published: April 2012

BackgroundSpindle cell lesions of the breast represent an interesting diagnostic challenge as they comprise a wide range of tumours that are rare. Differentiating dermatofibrosarcoma protuberans (DFSP) from other dermatofibromas using CD34 immunohistochemistry alone is difficult; therefore, fluorescence in situ hybridisation (FISH) analysis is often employed to identify typicalCOL1A1–PDGFBfusion or gene rearrangement. Although molecular confirmation of diagnosis is unnecessary in the majority of DFSP cases, the detection of chromosomal rearrangement is valuable in tumours that show unusual clinicopathological features as in this study the authors report a case of DFSP of breast that did not show any typical known molecular features.Methods and resultsMorphological and immunohistochemical study was highly suggestive of the diagnosis of DFSP. To further investigate this case, DNA copy number alterations were investigated by the 250 K Affymetrix SNP Mapping array. DNA analysis did not show any of the known translocations reported in DFSP or any known solid tumour category. However, in addition to copy number changes on chromosome 1, amplification of chromosome 7p which contains theepidermal growth factor receptor(EGFR) gene was observed. Results fromEGFRFISH showed an increase inEGFRgene to chromosome 7 ratio (3:1) suggesting amplification of theEGFRgene.ConclusionThis case of an unusual DFSP demonstrates that genomic interrogation provides additional potential targets such as a therapeutic avenue with anti-EGFRtherapies and shows the power of molecular characterisation of unusual tumours for a personalised medicine approach.
The prognostic significance of early stage lymph node positivity in operable invasive breast carcinoma: number or stage
Authors: Rakha Emad A , Morgan David , Macmillan Douglas 
Published: April 2012

AimThe earlier detection of breast cancer through mammographic screening has resulted in a shift in stage distribution with patients who are node-positive tending to present with a lower number of positive lymph nodes (LN). This study aims to assess the prognostic value of absolute number of positive nodes in the pN1 TNM stage (1–3 positive LN) and whether the prognostic value of the number of nodes in this clinically important stage justifies its consideration in management decisions.MethodsThis study is based on a large and well-characterised consecutive series of operable breast cancer (3491 cases), treated according to standard protocols in a single institution, with a long-term follow-up.ResultsLN stages and the absolute number of LN are associated with both breast cancer specific survival (BCSS) and distant metastasis free survival (DMFS). In the pN1 stage, patients with three positive LN (14% of pN1) show shorter BCSS (HR=1.9, (95% CI 1.3 to 2.6)) and shorter DMFS (HR=2.2, (95% CI 1.6 to 2.9)) when compared with one and/or two positive nodes. This effect is noted in the whole series as well as in different subgroups based on tumour size (pT1c and pT2), histological grade (grade 2 and 3), vascular invasion and oestrogen receptor status (both positive and negative). Multivariable analyses showed that three positive LN, compared with one and two positive LN, are an independent predictor of shorter BCSS and DMFS.ConclusionThe number of LN in the pN1 stage yielded potentially informative risk assignments with three positive LN providing an independent predictor of poorer outcome.
Prospective associations between meth/amphetamine (speed) and MDMA (ecstasy) use and depressive symptoms in secondary school students
Authors: Brière Frédéric N , Fallu Jean-Sébastien , Janosz Michel , Pagani Linda S 
Published: April 2012

BackgroundResearch has raised significant concern regarding the affective consequences of synthetic drug use. However, little evidence from well-controlled longitudinal studies exists on these consequences. The aim of this study was to determine whether use of meth/amphetamine (speed) and ±3,4-methylenedioxymethamphetamine (MDMA, ecstasy) is independently predictive of subsequent depressive symptoms in adolescents.MethodsA sample of 3880 adolescents from secondary schools in disadvantaged areas of Quebec, Canada, were followed over time (2003–2008). Logistic regression was used to test the association between meth/amphetamine and MDMA use in grade 10 (ages 15–16 years) and elevated depressive symptoms on an abridged Center for Epidemiologic Studies-Depression scale in grade 11, controlling for pre-existing individual and contextual characteristics.ResultsAfter adjustment, both MDMA use (OR 1.7, 95% CI 1.1 to 2.6) and meth/amphetamine use (OR 1.6, 95% CI 1.1 to 2.3) in grade 10 significantly increased the odds of elevated depressive symptoms in grade 11. These relationships did not vary by gender or pre-existing depressive symptoms. Increased risk was particularly observed in concurrent usage (OR 1.9, 95% CI 1.2 to 2.9).ConclusionsAdolescent use of meth/amphetamine and MDMA (particularly concurrent use) is independently associated with subsequent depressive symptoms. Further enquiry must determine whether these associations reflect drug-induced neurotoxicity and whether adolescence is a period of increased vulnerability to the hazards of synthetic drug exposure.
Novel TOPK Inhibitor HI-TOPK-032 Effectively Suppresses Colon Cancer Growth
Authors: Kim Dong Joon , Li Yan , Reddy Kanamata , Lee Mee-Hyun , Kim Myoung Ok , Cho Yong-Yeon , Lee Sung-Young , Kim Jong-Eun , Bode Ann M. , Dong Zigang 
Published: April 2012

Abstract
The serine-threonine mitogen-activated protein kinase kinase family member T-LAK cell–originated protein kinase (TOPK/PBK) is heavily involved in tumor development, cancer growth, apoptosis, and inflammation. Despite the identification of TOPK as a promising novel therapeutic target, no inhibitor of TOPK has yet been reported. In this study, we screened 36 drug candidates using an in vitro kinase assay and identified the novel TOPK inhibitor HI-TOPK-032. In vitro, HI-TOPK-032 strongly suppressed TOPK kinase activity but had little effect on extracellular signal–regulated kinase 1 (ERK1), c-jun—NH2—kinase 1, or p38 kinase activities. HI-TOPK-032 also inhibited anchorage-dependent and -independent colon cancer cell growth by reducing ERK-RSK phosphorylation as well as increasing colon cancer cell apoptosis through regulation of the abundance of p53, cleaved caspase-7, and cleaved PARP. In vivo, administration of HI-TOPK-032 suppressed tumor growth in a colon cancer xenograft model. Our findings therefore show that HI-TOPK-032 is a specific inhibitor of TOPK both in vitro and in vivo that may be further developed as a potential therapeutic against colorectal cancer. Cancer Res; 72(12); 3060–8. ©2012 AACR.
STEAP Proteins: From Structure to Applications in Cancer Therapy
Authors: Gomes Inês M. , Maia Cláudio J. , Santos Cecília R. 
Published: April 2012

Abstract
The human 6-transmembrane epithelial antigen of prostate (STEAP) family comprises STEAP1, STEAP2, STEAP3, and STEAP4. All of these proteins are unique to mammals and share an innate activity as metalloreductases, indicating their importance in metal metabolism. Overall, they participate in a wide range of biologic processes, such as molecular trafficking in the endocytic and exocytic pathways and control of cell proliferation and apoptosis. STEAP1 and STEAP2 are overexpressed in several types of human cancers, namely prostate, bladder, colon, pancreas, ovary, testis, breast, cervix, and Ewing sarcoma, but their clinical significance and role in cancer cells are not clear. Still, their localization in the cell membrane and differential expression in normal and cancer tissues make STEAP proteins potential candidates as biomarkers of several cancers, as well as potential targets for new immunotherapeutic strategies for disease attenuation or treatment. This review brings together the current knowledge about each STEAP protein, giving an overview of the roles of this family of proteins in human physiology and disease, and analyzes their potential as immunotherapeutic agents in cancer research. Mol Cancer Res; 10(5); 573–87. ©2012 AACR.
The Synaptic Proteins β-Neurexin and Neuroligin Synergize With Extracellular Matrix-Binding Vascular Endothelial Growth Factor A During Zebrafish Vascular Development
Authors: Rissone Alberto , Foglia Efrem , Sangiorgio Lorenzo , Cermenati Solei , Nicoli Stefania , Cimbro Simona , Beltrame Monica , Bussolino Federico , Cotelli Franco , Arese Marco 
Published: April 2012

Objective—
The goal of this study was to determine the in vivo functions of the synaptic proteins neurexins and neuroligins in embryonic vascular system development using zebrafish as animal model.

Methods and Results—

In the present study, we show that the knockdown of the α-form of neurexin 1a induces balance defects and reduced locomotory activity, whereas β-neurexin 1a and neuroligin 1 morphants present defects in sprouting angiogenesis and vascular remodeling, in particular in the caudal plexus and subintestinal vessels. Coinjection of low doses of morpholinos for β-neurexin 1a and neuroligin 1 together or in combination with morpholinos targeting the ­heparin-­binding isoforms of vascular endothelial growth factor A (encoded by the
VEGFAb
gene) recapitulates the observed abnormalities, suggesting synergistic activity of these molecules. Similar coinjection experiments with morpholinos, targeting the enzyme heparan sulfate 6-­O-­sulfotransferase 2, confirm the presence of a functional correlation between extracellular matrix maturation and β-neurexin 1a or neuroligin 1.

Conclusion—
Our data represent the first in vivo evidence of the role of neurexin and neuroligin in embryonic blood vessel formation and provide insights into their mechanism of action
Intense Physiological Light Upregulates Vascular Endothelial Growth Factor and Enhances Choroidal Neovascularization via Peroxisome Proliferator-Activated Receptor γ Coactivator-1α in Mice
Authors: Ueta Takashi , Inoue Tatsuya , Yuda Kentaro , Furukawa Takahisa , Yanagi Yasuo , Tamaki Yasuhiro 
Published: April 2012

Objectives—
Toxicity of intense light to facilitate the development of neovascular age-related macular degeneration has been a health concern although the mechanism remains unclear.

Methods and Results—

Effects of intense, but within physiological range, light on retinal pigment epithelium, a major pathogenic origin of age-related macular degeneration were studied in mice. Intense physiological light upregulated vascular endothelial growth factor (VEGF) expression in retinal pigment epithelium, independent of circadian rhythm, which resulted in enhancement of choroidal neovascularization. In
rd1/rd1
mice or
Crx
−/−
mice that do not possess outer segment structure, light exposure did not induce VEGF, indicating that VEGF upregulation by light depended on increased outer segment phagocytosis by retinal pigment epithelium. In retinal pigment epithelium cells phagocytosing increased amount of outer segment, peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) not hypoxia-inducible factor-1α was induced, leading to VEGF upregulation. The VEGF upregulation and choroidal neovascularization enhancement were abrogated in
PGC-1α
−/−
mice and estrogen-related receptor-α
−/−
mice, indicating the involvement of PGC-1α/estrogen-related receptor-α pathway.

Conclusions—
Intense physiological light is involved in choroidal neovascularization through excess outer segment phagocy-tosis and VEGF upregulation mediated by PGC-1α in vivo.
Role of Src Tyrosine Kinases in Experimental Pulmonary Hypertension
Authors: Pullamsetti Soni Savai , Berghausen Eva Maria , Dabral Swati , Tretyn Aleksandra , Butrous Elsa , Savai Rajkumar , Butrous Ghazwan , Dahal Bhola Kumar , Brandes Ralf P. , Ghofrani Hossein Ardeschir , Weissmann Norbert , Grimminger Friedrich , Seeger Werner , Rosenkranz Stephan , Schermuly Ralph Theo 
Published: April 2012

Objective—
Pulmonary arterial hypertension is a progressive pulmonary vascular disorder with high morbidity and mortality. Compelling evidence suggests that receptor tyrosine kinases, such as platelet-derived growth factor (PDGF) are closely involved in the pathogenesis of pulmonary arterial hypertension. We investigated the effects of 2 novel PDGF inhibitors, nilotinib/AMN107 (Abl kinases/PDGF receptor inhibitor) and dasatinib/BMS-354825 (Abl kinases/PDGF receptor/Src inhibitor), on the proliferation and migration of pulmonary arterial smooth muscle cells (PASMCs) and on the hemodynamics and pulmonary vascular remodeling in experimental pulmonary hypertension, and determined the expression and regulation of Src family kinases.

Methods and Results—
Human PASMCs were stimulated by PDGF alone or multiple growth factors to induce proliferation and migration in vitro. Dasatinib (0.03 μmol/L), nilotinib (0.3 μmol/L), and imatinib (1 μmol/L) potently inhibited PDGF-induced signal transducer and activator of transcription 3 and Akt phosphorylation. All 3 inhibitors decreased PDGF-induced proliferation, cell cycle gene regulation, and migration. In contrast, only dasatinib inhibited multiple growth factor–induced PASMC proliferation, and this was associated with the inhibition of Src phosphorylation. Combination of specific Src inhibitors (phosphoprotein phosphatase 1, phosphoprotein phosphatase 2) with either imatinib or nilotinib reduced multiple growth factor–induced proliferation to a similar extent as dasatinib. Importantly, Src phosphorylation increased in pulmonary arterial hypertension PASMCs compared with control PASMCs. Finally, in vivo dasatinib (15 mg/kg per body weight) treatment caused a complete reversal of pulmonary vascular remodeling and achieved similar effectiveness as imatinib (100 mg/kg per body weight) in both monocrotaline- and hypoxia-induced pulmonary hypertension models.

Conclusion—
We suggest that dual inhibition of PDGF receptor and Src kinases potently inhibits mitogenic and motogenic responses to growth factors in PASMCs and pulmonary vascular remodeling in vivo so that dual inhibition may represent an alternative therapeutic approach for pulmonary arterial hypertension.