bluuebunny/crossref_metadata_2025_split huggingface dataset:
One-Step Sequence- and Ligation-Independent Cloning as a Rapid and Versatile Cloning Method for Functional Genomics Studies
Authors: Jeong Jae-Yeon , Yim Hyung-Soon , Ryu Ji-Young , Lee Hyun Sook , Lee Jung-Hyun , Seen Dong-Seung , Kang Sung Gyun 
Published: May 2012

ABSTRACT
We developed one-step sequence- and ligation-independent cloning (SLIC) as a simple, cost-effective, time-saving, and versatile cloning method. Highly efficient and directional cloning can be achieved by direct bacterial transformation 2.5 min after mixing any linearized vector, an insert(s) prepared by PCR, and T4 DNA polymerase in a tube at room temperature.
Decreased Toxicity of Bacillus thuringiensis subsp. israelensis to Mosquito Larvae after Contact with Leaf Litter
Authors: Tetreau Guillaume , Stalinski Renaud , Kersusan Dylann , Veyrenc Sylvie , David Jean-Philippe , Reynaud Stéphane , Després Laurence 
Published: May 2012

ABSTRACTBacillus thuringiensissubsp.israelensisis a bacterium producing crystals containing Cry and Cyt proteins, which are toxic for mosquito larvae. Nothing is known about the interaction between crystal toxins and decaying leaf litter, which is a major component of several mosquito breeding sites and represents an important food source. In the present work, we investigated the behavior ofB. thuringiensissubsp.israelensistoxic crystals sprayed on leaf litter. In the presence of leaf litter, a 60% decrease in the amount of Cyt toxin detectable by immunology (enzyme-linked immunosorbent assays [ELISAs]) was observed, while the respective proportions of Cry toxins were not affected. The toxicity of Cry toxins towardAedes aegyptilarvae was not affected by leaf litter, while the synergistic effect of Cyt toxins on allB. thuringiensissubsp.israelensisCry toxins was decreased by about 20% when mixed with leaf litter. The toxicity of two commercialB. thuringiensissubsp.israelensisstrains (VectoBac WG and VectoBac 12AS) and a laboratory-producedB. thuringiensissubsp.israelensisstrain decreased by about 70% when mixed with leaf litter. Taken together, these results suggest that Cyt toxins interact with leaf litter, resulting in a decreased toxicity ofB. thuringiensissubsp.israelensisin litter-rich environments and thereby dramatically reducing the efficiency of mosquitocidal treatments.
A Gain-of-Function Mutation in Gating of Corynebacterium glutamicum NCgl1221 Causes Constitutive Glutamate Secretion
Authors: Nakayama Yoshitaka , Yoshimura Kenjiro , Iida Hidetoshi 
Published: May 2012

ABSTRACT

The A-to-V mutation at position 111 (A111V) in the mechanosensitive channel NCgl1221 (MscCG) causes constitutive glutamate secretion in
Corynebacterium glutamicum
. Patch clamp experiments revealed that NCgl1221 (A111V) had a significantly smaller gating threshold than the wild-type counterpart and displayed strong hysteresis, suggesting that the gain-of-function mutation in the gating of NCgl1221 leads to the oversecretion of glutamate.
Microbial Quality of Tropical Inland Waters and Effects of Rainfall Events
Authors: Santiago-Rodriguez Tasha M. , Tremblay Raymond L. , Toledo-Hernandez Carlos , Gonzalez-Nieves Joel E. , Ryu Hodon , Santo Domingo Jorge W. , Toranzos Gary A. 
Published: May 2012

ABSTRACT

Novel markers of fecal pollution in tropical waters are needed since conventional methods recommended for other geographical regions may not apply. To address this, the prevalence of thermotolerant coliforms, enterococci, coliphages, and enterophages was determined by culture methods across a watershed. Additionally, human-, chicken-, and cattle-specific PCR assays were used to identify potential fecal pollution sources in this watershed. An enterococcus quantitative PCR (qPCR) assay was tested and correlated with culture methods at three sites since water quality guidelines could incorporate this technique as a rapid detection method. Various rainfall events reported before sample collection at three sites were considered in the data analyses. Thermotolerant coliforms, enterococci, coliphages, and enterophages were detected across the watershed. Human-specific
Bacteroides
bacteria, unlike the cattle- and chicken-specific bacteria, were detected mostly at sites with the corresponding fecal impact. Enterococci were detected by qPCR as well, but positive correlations with the culture method were noted at two sites, suggesting that either technique could be used. However, no positive correlations were noted for an inland lake tested, suggesting that qPCR may not be suitable for all water bodies. Concentrations of thermotolerant coliforms and bacteriophages were consistently lower after rainfall events, pointing to a possible dilution effect. Rainfall positively correlated with enterococci detected by culturing and qPCR, but this was not the case for the inland lake. The toolbox of methods and correlations presented here could be potentially applied to assess the microbial quality of various water types.
SPO71
Mediates Prospore Membrane Size and Maturation in Saccharomyces cerevisiae
Authors: Parodi Emily M. , Baker Crystal S. , Tetzlaff Cayla , Villahermosa Sasha , Huang Linda S. 
Published: May 2012

ABSTRACT

The mechanisms that control the size and shape of membranes are not well understood, despite the importance of these structures in determining organelle and cell morphology. The prospore membrane, a double lipid bilayer that is synthesized
de novo
during sporulation in
S. cerevisiae
, grows to surround the four meiotic products. This membrane determines the shape of the newly formed spores and serves as the template for spore wall deposition. Ultimately, the inner leaflet of the prospore membrane will become the new plasma membrane of the cell upon germination. Here we show that Spo71, a pleckstrin homology domain protein whose expression is induced during sporulation, is critical for the appropriate growth of the prospore membrane. Without
SPO71
, prospore membranes surround the nuclei but are abnormally small, and spore wall deposition is disrupted. Sporulating
spo71
Δ cells have prospore membranes that properly localize components to their growing leading edges yet cannot properly localize septin structures. We also found that
SPO71
genetically interacts with
SPO1
, a gene with homology to the phospholipase B gene that has been previously implicated in determining the shape of the prospore membrane. Together, these results show that
SPO71
plays a critical role in prospore membrane development.
Functional Analyses of Mycobacterial Lipoprotein Diacylglyceryl Transferase and Comparative Secretome Analysis of a Mycobacterial
lgt
Mutant
Authors: Tschumi Andreas , Grau Thomas , Albrecht Dirk , Rezwan Mandana , Antelmann Haike , Sander Peter 
Published: May 2012

ABSTRACT

Preprolipopoprotein diacylglyceryl transferase (Lgt) is the gating enzyme of lipoprotein biosynthesis, and it attaches a lipid structure to the N-terminal part of preprolipoproteins. Using Lgt from
Escherichia coli
in a BLASTp search, we identified the corresponding Lgt homologue in
Mycobacterium tuberculosis
and two homologous (MSMEG_3222 and MSMEG_5408) Lgt in
Mycobacterium smegmatis. M. tuberculosis lgt
was shown to be essential, but an
M. smegmatis
ΔMSMEG_3222 mutant could be generated. Using Triton X-114 phase separation and [
14
C]palmitic acid incorporation, we demonstrate that MSMEG_3222 is the major Lgt in
M. smegmatis
. Recombinant
M. tuberculosis
lipoproteins Mpt83 and LppX are shown to be localized in the cell envelope of parental
M. smegmatis
but were absent from the cell membrane and cell wall in the
M. smegmatis
ΔMSMEG_3222 strain. In a proteomic study, 106 proteins were identified and quantified in the secretome of wild-type
M. smegmatis
, including 20 lipoproteins. All lipoproteins were secreted at higher levels in the ΔMSMEG_3222 mutant. We identify the major Lgt in
M. smegmatis
, show that lipoproteins lacking the lipid anchor are secreted into the culture filtrate, and demonstrate that
M. tuberculosis lgt
is essential and thus a validated drug target.
Identification of a Third Osmoprotectant Transport System, the OsmU System, in Salmonella enterica
Authors: Frossard Stephen M. , Khan Aftab A. , Warrick Eric C. , Gately Jonathan M. , Hanson Andrew D. , Oldham Michael L. , Sanders David Avram , Csonka Laszlo N. 
Published: May 2012

ABSTRACTThe growth ofSalmonella entericaserovar Typhimurium mutants lacking the ProP and ProU osmoprotectant transport systems is stimulated by glycine betaine in high-osmolarity media, suggesting that this organism has an additional osmoprotectant transport system. Bioinformatic analysis revealed that the genome of this organism contains a hitherto-unidentified operon, designatedosmU, consisting of four genes whose products show high similarity to ABC-type transport systems for osmoprotectants in other bacteria. TheosmUoperon was inactivated by a site-directed deletion, which abolished the ability of glycine betaine to alleviate the inhibitory effect of high osmolarity and eliminated the accumulation of [14C]glycine betaine and [14C]choline-O-sulfate in high-osmolarity media in a strain lacking the ProP and ProU systems. Although the OsmU system can take up glycine betaine and choline-O-sulfate, these two osmoprotectants are recognized at low affinity by this transporter, suggesting that there might be more efficient substrates that are yet to be discovered. The transcription ofosmUis induced 23-fold by osmotic stress (0.3 M NaCl). TheosmUoperon is present in the genomes of a number ofEnterobacteriaceae, and orthologs of the OsmU system can be recognized in a wide variety ofBacteriaandArchaea. The structure of the periplasmic binding protein component of this transporter, OsmX, was modeled on the crystallographic structure of the glycine betaine-binding protein ProX ofArchaeoglobus fulgidus; the resultant model indicated that the amino acids that constitute substrate-binding site, including an “aromatic cage” made up of four tyrosines, are conserved between these two proteins.
Extracellular Identification of a Processed Type II ComR/ComS Pheromone of
Streptococcus mutans
Authors: Khan Rabia , Rukke Håkon V. , Ricomini Filho Antonio Pedro , Fimland Gunnar , Arntzen Magnus Ø. , Thiede Bernd , Petersen Fernanda C. 
Published: May 2012

ABSTRACT

The competence-stimulating peptide (CSP) and the
sigX
-inducing peptide (XIP) are known to induce
Streptococcus mutans
competence for genetic transformation. For both pheromones, direct identification of the native peptides has not been accomplished. The fact that extracellular XIP activity was recently observed in a chemically defined medium devoid of peptides, as mentioned in an accompanying paper (

K. Desai, L. Mashburn-Warren, M. J. Federle, and D. A. Morrison, J. Bacteriol.
194
:3774–3780, 2012

), provided ideal conditions for native XIP identification. To search for the XIP identity, culture supernatants were filtered to select for peptides of less than 3 kDa, followed by C
18
extraction. One peptide, not detected in the supernatant of a
comS
deletion mutant, was identified by tandem mass spectrometry (MS/MS) fragmentation as identical to the ComS C-terminal sequence GLDWWSL. ComS processing did not require Eep, a peptidase involved in processing or import of bacterial small hydrophobic peptides, since
eep
deletion had no inhibitory effect on XIP production or on synthetic XIP response. We investigated whether extracellular CSP was also produced. A reporter assay for CSP activity detection, as well as MS analysis of supernatants, revealed that CSP was not present at detectable levels. In addition, a mutant with deletion of the CSP-encoding gene
comC
produced endogenous XIP levels similar to those of a nondeletion mutant. The results indicate that XIP pheromone production is a natural phenomenon that may occur in the absence of natural CSP pheromone activity and that the heptapeptide GLDWWSL is an extracellular processed form of ComS, possibly the active XIP pheromone. This is the first report of direct identification of a ComR/ComS pheromone.
The natural history and predictors of persistent problem behaviours in cystic fibrosis: a multicentre, prospective study
Authors: Sheehan Jane , Massie John , Hay Margaret , Jaffe Adam , Glazner Judith , Armstrong David , Hiscock Harriet 
Published: May 2012

BackgroundIn cystic fibrosis (CF), problems with sleep, eating/mealtime behaviours, physiotherapy adherence and parental mental health issues are common, yet their natural history and the extent of service use to address them are unknown.ObjectiveFollow up the 2007 cohort to determine: (1) prevalence of child sleep, eating/mealtime behaviours, physiotherapy adherence, and externalising/internalising problem behaviours and primary caregiver mental health status after a 3-year period; (2) natural history of child behaviours; (3) potentially modifiable predictors of persistent problems; and (4) service use for behaviours.DesignProspective cohort. Setting: Royal Children's Hospital, Monash Medical Centre and Sydney Children's Hospital (Australia) CF clinics.ParticipantsCaregivers, of children aged 3–8 years, who completed the baseline questionnaire.Main outcome measuresChild sleep, eating/mealtime behaviours, adherence with therapy and externalising and internalising behaviours. Predictors: parenting style (low warmth), caregiver mental health status and sleep quality at baseline.Results102 of 116 (88%) families participated. The prevalence of sleep and eating/mealtime problems at follow-up was similar to baseline. The prevalence of caregiver mental health symptoms remained higher than the community data. 71 out of 102 (70%) of the children experienced at least one persistent behaviour problem. Caregiver mental health difficulties predicted persistent child moderate to severe sleep problems (adjusted OR 6.5, 95% CI 1.2 to 36.2, p=0.03) and their persistently poor adherence to the physiotherapy regimen (adjusted OR 3.5, 95% CI 1.3 to 9.2, p=0.01).ConclusionsChild problem behaviours are common in children with CF, persist and are often predicted by the mental health of the parent. Routine surveillance for and management of child problem behaviours are recommended.
Sports injuries and illnesses during the second Asian Beach Games
Authors: Al-Shaqsi Sultan , Al-Kashmiri Ammar , Al-Risi Ahmed , Al-Mawali Suleiman 
Published: May 2012

BackgroundPrevention of sport injuries and illnesses is a focus for epidemiological surveillance.ObjectivesTo record and analyse all sports injuries and illnesses registered during the second Asian Beach Games.DesignA descriptive epidemiological study using the International Olympic Committee Surveillance system to register injuries and illnesses during the second Asian Beach Games.MethodsThe second Asian Beach Games hosted 1132 athletes from 43 countries competing in 14 beach sports. All National Olympic Committees' physicians of the participating teams were invited to report all injuries and illnesses. In addition, medical officers at the different Olympic venues and the main Olympic village reported injuries and illnesses treated at the clinics on a daily basis.ResultsA total of 177 injuries were reported equating to an incidence rate of 156.4 per 1000 registered athletes. Tent pegging recorded the highest incidence of injuries with 357 per 1000 registered athletes. The most prevalent injuries were in the foot/toe with 14.1% of all reported injuries. The majority of injuries were incurred during competition (75.4%). In addition, the most common mechanism of injury was contact with another athlete (n=42, 23.7%) and combined sudden and gradual overuse contributed to 30% of the total injury burden. Furthermore, 118 illnesses were reported resulting in an incidence rate of 104.2 illnesses per 1000 registered athletes. The most affected system was the respiratory tract (39.1%) with infection being the most common cause (n=33, 38.0%). The incidence of injury and illness differed significantly among the 14 sports.ConclusionThe data indicate that the risk of injury from beach games is sport dependant. This means that any preventive measures have to be tailored for each discipline. Furthermore, the study showed that respiratory infections are the commonest illness in beach sports and therefore, event organisers should focus improving public health measures and hygiene awareness.
The use of recombinant activated factor VII (rFVIIa) in the management of patients with major haemorrhage in military hospitals over the last 5 years
Authors: Smith J E 
Published: May 2012

IntroductionRecombinant activated factor VII (rFVIIa) has been used in the management of traumatic haemorrhage for a decade. Anecdotally, its use is decreasing. The aim of this study was to define the use of rFVIIa in UK deployed military hospitals over the last 5 years.MethodsA retrospective database review was performed, using the Joint Theatre Trauma Registry, for the period January 2006 to June 2011. Data collected included use of rFVIIa, injury severity score (ISS), survival and injury pattern. The temporal trend of rFVIIa use, taking into account the number of severely injured patients presenting during each time period, was then analysed.ResultsDuring the period January 2006 to June 2011, 156 injured patients received rFVIIa. 146 of these (94%) had an ISS >15; there were 45 fatalities. The median ISS among the group receiving rFVIIa was 30, and 20 patients had an ISS in the range 60–75. There was a significant reduction in the use of rFVIIa in the second half of 2010 and the first half of 2011, compared with the previous 12-month period.ConclusionThe use of rFVIIa in UK deployed military hospitals has declined since 2010, which is likely due to a combination of factors, including a change in resuscitation practice in these units, and a change in emphasis of manufacturer's guidance.
Macrolide resistance testing and molecular subtyping ofTreponema pallidumstrains from southern Africa
Authors: Müller Etienne E , Paz-Bailey Gabriela , Lewis David A 
Published: May 2012

ObjectivesTo determine whether the 23S ribosomal RNA (rRNA) A2058G and A2059G mutations that confer macrolide resistance are present among southern African strains ofTreponema pallidumand to determine their subtype distribution.Methods117 genital ulcer specimens, collected between March 2005 and April 2010 in South Africa and Lesotho and previously determined to be positive forT pallidumDNA by molecular testing, were retested using a commercial real-time PCR assay. Those specimens that were still positive forT pallidumDNA were screened for the macrolide resistance-encoding point mutations in the 23S rRNA gene using rapid PCR-based restriction digest assays. Molecular characterisation of two variable treponemal genes,arpandtpr, was used to subtype theT pallidumstrains.Results1 of 100T pallidum-positive specimens, collected in Lesotho, contained the A2058G macrolide resistance-encoding 23S rRNA gene mutation, whereas the A2059G mutation was absent. It was possible to fully type 97/100 of allT pallidumDNA-positive samples. A total of ninearprepeat sizes, ninetprpatterns and a combined total of 20 subtypes were identified. Overall, the most common subtypes were 14d (32%), followed by 17d (12%), 14a (10%), 14b (8%), 22b (6%) and 14i (5%). Subtypes 14d and 14a were the predominant subtypes in samples from South Africa (43%) and Lesotho (22%), respectively.ConclusionsMacrolide resistance amongT pallidumstrains appears to be uncommon in southern Africa. Although a high degree of genetic heterogeneity was observed among the strains tested,T pallidumsubtype 14d appears to be the predominant circulating strain.
Vandetanib, an Inhibitor of VEGF Receptor-2 and EGF Receptor, Suppresses Tumor Development and Improves Prognosis of Liver Cancer in Mice
Authors: Inoue Kinya , Torimura Takuji , Nakamura Toru , Iwamoto Hideki , Masuda Hiroshi , Abe Mitsuhiko , Hashimoto Osamu , Koga Hironori , Ueno Takato , Yano Hirohisa , Sata Michio 
Published: May 2012

Abstract
Purpose: VEGF, EGF, and TGF-α are expressed in hepatocellular carcinomas (HCC) and play a role in its growth. Vandetanib, a multikinase inhibitor, suppresses the phosphorylation of VEGF receptor 2 (VEGFR-2) and EGF receptor (EGFR). The aim of this study was to clarify the antitumor effect of vandetanib in mouse HCCs.
Experimental Design: We evaluated the effects of vandetanib on proliferation of human umbilical vein endothelial cells (HUVEC) and three hepatoma cell lines, as well as the phosphorylation of VEGFR-2 and EGFR in these cells. Mice were implanted with hepatoma cells subcutaneously or orthotopically in the liver and treated with 50 or 75 mg/kg vandetanib. We analyzed the effects of treatment on tumor cell proliferation and apoptosis, vessel density, phosphorylation of VEGFR-2 and EGFR, and production of VEGF, TGF-α, and EGF in tumor tissues. Adverse events on vandetanib administration were also investigated.
Results: Vandetanib suppressed phosphorylation of VEGFR-2 in HUVECs and EGFR in hepatoma cells and inhibited cell proliferation. In tumor-bearing mice, vandetanib suppressed phosphorylation of VEGFR-2 and EGFR in tumor tissues, significantly reduced tumor vessel density, enhanced tumor cell apoptosis, suppressed tumor growth, improved survival, reduced number of intrahepatic metastases, and upregulated VEGF, TGF-α, and EGF in tumor tissues. Treatment with vandetanib was not associated with serious adverse events, including alanine aminotransferase abnormality, bone marrow suppression, or body weight loss.
Conclusions: The antitumor effects of vandetanib in mice suggest that it is a potentially suitable and safe chemotherapeutic agent for HCCs. Clin Cancer Res; 18(14); 3924–33. ©2012 AACR.
MLN0905, a Small-Molecule PLK1 Inhibitor, Induces Antitumor Responses in Human Models of Diffuse Large B-cell Lymphoma
Authors: Shi Judy Quiju , Lasky Kerri , Shinde Vaishali , Stringer Bradley , Qian Mark G. , Liao Debra , Liu Ray , Driscoll Denise , Nestor Michelle Tighe , Amidon Benjamin S. , Rao Youlan , Duffey Matt O. , Manfredi Mark G. , Vos Tricia J. , Amore Natalie D' , Hyer Marc L. 
Published: May 2012

Abstract
Diffuse large B-cell lymphoma (DLBCL) is the most common of the non–Hodgkin lymphomas, accounting for up to 30% of all newly diagnosed lymphoma cases. Current treatment options for this disease are effective, but not always curative; therefore, experimental therapies continue to be investigated. We have discovered an experimental, potent, and selective small-molecule inhibitor of PLK1, MLN0905, which inhibits cell proliferation in a broad range of human tumor cells including DLBCL cell lines. In our report, we explored the pharmacokinetic, pharmacodynamic, and antitumor properties of MLN0905 in DLBCL xenograft models grown in mice. These studies indicate that MLN0905 modulates the pharmacodynamic biomarker phosphorylated histone H3 (pHisH3) in tumor tissue. The antitumor activity of MLN0905 was evaluated in three human subcutaneous DLBCL xenograft models, OCI LY-10, OCI LY-19, and PHTX-22L (primary lymphoma). In each model, MLN0905 yielded significant antitumor activity on both a continuous (daily) and intermittent dosing schedule, underscoring dosing flexibility. The antitumor activity of MLN0905 was also evaluated in a disseminated xenograft (OCI LY-19) model to better mimic human DLBCL disease. In the disseminated model, MLN0905 induced a highly significant survival advantage. Finally, MLN0905 was combined with a standard-of-care agent, rituximab, in the disseminated OCI LY-19 xenograft model. Combining rituximab and MLN0905 provided both a synergistic antitumor effect and a synergistic survival advantage. Our findings indicate that PLK1 inhibition leads to pharmacodynamic pHisH3 modulation and significant antitumor activity in multiple DLBCL models. These data strongly suggest evaluating PLK1 inhibitors as DLBCL anticancer agents in the clinic. Mol Cancer Ther; 11(9); 2045–53. ©2012 AACR.