bluuebunny/crossref_metadata_2025_split huggingface dataset:
Protein body formation in stable transgenic tobacco expressing elastin-like polypeptide and hydrophobin fusion proteins
Authors: Gutiérrez Sonia P , Saberianfar Reza , Kohalmi Susanne E , Menassa Rima 
Published: May 2013

Abstract

Background
Plants are recognized as an efficient and inexpensive system to produce valuable recombinant proteins. Two different strategies have been commonly used for the expression of recombinant proteins in plants: transient expression mediated by Agrobacterium; or stable transformation of the plant genome. However, the use of plants as bioreactors still faces two main limitations: low accumulation levels of some recombinant proteins and lack of efficient purification methods. Elastin-like polypeptide (ELP), hydrophobin I (HFBI) and Zera® are three fusion partners found to increase the accumulation levels of recombinant proteins and induce the formation of protein bodies (PBs) in leaves when targeted to the endoplasmic reticulum (ER) in transient expression assays. In this study the effects of ELP and HFBI fusion tags on recombinant protein accumulation levels and PB formation was examined in stable transgenic Nicotiana tabacum.

Results
The accumulation of recombinant protein and PB formation was evaluated in two cultivars of Nicotiana tabacum transformed with green fluorescent protein (GFP) fused to ELP or HFBI, both targeted and retrieved to the ER. The ELP and HFBI tags increased the accumulation of the recombinant protein and induced the formation of PBs in leaves of stable transgenic plants from both cultivars. Furthermore, these tags induced the formation of PBs in a concentration-dependent manner, where a specific level of recombinant protein accumulation was required for PBs to appear. Moreover, agro-infiltration of plants accumulating low levels of recombinant protein with p19, a suppressor of post-transcriptional gene silencing (PTGS), increased accumulation levels in four independent transgenic lines, suggesting that PTGS might have caused the low accumulation levels in these plants.

Conclusion
The use of ELP and HFBI tags as fusion partners in stable transgenic plants of tobacco is feasible and promising. In a constitutive environment, these tags increase the accumulation levels of the recombinant protein and induce the formation of PBs regardless of the cultivar used. However, a specific level of recombinant protein accumulation needs to be reached for PBs to form.
Economic evaluation of agomelatine relative to other antidepressants for treatment of major depressive disorders in Greece
Authors: Maniadakis Nikos , Kourlaba Georgia , Mougiakos Theodoros , Chatzimanolis Ioannis , Jonsson Linus 
Published: May 2013

AbstractBackgroundMajor depressive disorder (MDD) constitutes an important public health problem, as it is highly prevalent in the industrialized world and it is associated with substantial economic consequences for patients, health care providers, insurance and social security organizations and employers. To conduct an economic evaluation comparing agomelatine with other commonly used alternatives for treating patients with major depressive disorder (MDD) in Greece.MethodsAn existing international Markov model designed to evaluate the cost-effectiveness of agomelatine was adapted to the Greek setting. It reflects six different health states, in which patients may move on a monthly basis. The analysis was undertaken from a societal perspective. Transition probabilities, utilities and costs assigned to each health state were extracted from the published literature, government sources and expert opinion. Data reflects the year 2012 and was discounted using a rate of 3.5%. Probabilistic analysis was undertaken to deal with uncertainty.ResultsBase case analyses revealed that agomelatine is a dominant therapy for MDD relative to escitalopram, fluoxetine and sertraline, and it appeared to be cost-effective compared to venlafaxine (ICER: €547/QALY). Agomelatine remained a dominant treatment against generic sertraline and fluoxetine, and it appeared to be a cost-effective alternative compared to generic venlafaxine and escitalopram (ICER: €1,446/QALY and €3,303/QALY, respectively). Excluding the indirect cost from the analysis, agomelatine remained a cost-effective alternative over all comparators. In the probabilistic sensitivity analysis agomelatine was dominant in 44.5%, 89.6%, 70.6% and 84.6% of simulated samples against branded venlafaxine, escitalopram, fluoxetine and sertraline, respectively.ConclusionThe present evaluation indicates that agomelatine is either a dominant or a cost-effective alternative relative to branded or generic alternatives, in Greece.
A versatile, efficient strategy for assembly of multi-fragment expression vectors in Saccharomyces cerevisiae using 60 bp synthetic recombination sequences
Authors: Kuijpers Niels GA , Solis-Escalante Daniel , Bosman Lizanne , van den Broek Marcel , Pronk Jack T , Daran Jean-Marc , Daran-Lapujade Pascale 
Published: May 2013

AbstractBackgroundIn vivorecombination of overlapping DNA fragments for assembly of large DNA constructs in the yeastSaccharomyces cerevisiaeholds great potential for pathway engineering on a small laboratory scale as well as for automated high-throughput strain construction. However, the currentin vivoassembly methods are not consistent with respect to yields of correctly assembled constructs and standardization of parts required for routine laboratory implementation has not been explored. Here, we present and evaluate an optimized and robust method forin vivoassembly of plasmids from overlapping DNA fragments inS. cerevisiae.ResultsTo minimize occurrence of misassembled plasmids and increase the versatility of the assembly platform, two main improvements were introduced; i) the essential elements of the vector backbone (yeast episome and selection marker) were disconnected and ii) standardized 60 bp synthetic recombination sequences non-homologous with the yeast genome were introduced at each flank of the assembly fragments. These modifications led to a 100 fold decrease in false positive transformants originating from the backbone as compared to previous methods. Implementation of the 60 bp synthetic recombination sequences enabled high flexibility in the design of complex expression constructs and allowed for fast and easy construction of all assembly fragments by PCR. The functionality of the method was demonstrated by the assembly of a 21 kb plasmid out of nine overlapping fragments carrying six glycolytic genes with a correct assembly yield of 95%. The assembled plasmid was shown to be a high fidelity replica of thein silicodesign and all glycolytic genes carried by the plasmid were proven to be functional.ConclusionThe presented method delivers a substantial improvement for assembly of multi-fragment expression vectors inS. cerevisiae. Not only does it improve the efficiency ofin vivoassembly, but it also offers a versatile platform for easy and rapid design and assembly of synthetic constructs. The presented method is therefore ideally suited for the construction of complex pathways and for high throughput strain construction programs for metabolic engineering purposes. In addition its robustness and ease of use facilitate the construction of any plasmid carrying two or more genes.
Growth hormone response to submaximal doses of ghrelin remains unchanged during the follicular phase of the cycle
Authors: Messini Christina I , Dafopoulos Konstantinos , Malandri Maria , Georgoulias Panagiotis , Anifandis George , Messinis Ioannis E 
Published: May 2013

Abstract

Background
Previous data have shown that ghrelin-induced growth hormone (GH) secretion is augmented in women by exogenous but not by endogenous estrogens. The purpose of this study was to examine the response of GH to low-dose scheme of ghrelin administration in relation to physiological changes in estradiol levels during the normal menstrual cycle.

Methods
Ten normally cycling women were studied in two menstrual cycles. Two consecutive dosages of ghrelin (0.15 μg/kg and 0.30 μg/kg) were injected intravenously at 0 and 90 min in the early and late follicular phases of one cycle. Saline was injected in the preceding cycle. Blood samples were taken at −15, 0, 30, 60, 90, 120, 150 and 180 min. The GH response was assessed.

Results
Serum estradiol concentrations were significantly higher in the late than in the early follicular phase. After ghrelin, but not after saline administration, plasma ghrelin and serum GH levels increased significantly in both phases, peaking at 30 min and 120 min. The peak value at 120 min was significantly higher than at 30 min (P<0.001). There were no significant differences in ghrelin and GH levels between the two phases at all time points.

Conclusions
The present results show no difference in GH response to two consecutive submaximal doses of ghrelin between the early and the late follicular phase of the cycle. It is suggested that estradiol is not possibly involved in the physiological process that regulates ghrelin-induced GH secretion in women during the normal menstrual cycle.
Delayed brain ischemia tolerance induced by electroacupuncture pretreatment is mediated via MCP-induced protein 1
Authors: Jin Zhuqing , Liang Jian , Wang Jing , Kolattukudy Pappachan E 
Published: May 2013

Abstract

Background
Emerging studies have demonstrated that pretreatment with electroacupuncture (EA) induces significant tolerance to focal cerebral ischemia. The present study seeks to determine the involvement of monocyte chemotactic protein-induced protein 1 (MCPIP1), a recently identified novel modulator of inflammatory reactions, in the cerebral neuroprotection conferred by EA pretreatment in the animal model of focal cerebral ischemia and to elucidate the mechanisms of EA pretreatment-induced ischemic brain tolerance.

Methods
Twenty-four hours after the end of the last EA pretreatment, focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) for 90 minutes in male C57BL/6 mice and MCPIP1 knockout mice. Transcription and expression of MCPIP1 gene was monitored by qRT-PCR, Western blot and immunohistochemistry. The neurobehavioral scores, infarction volumes, proinflammatory cytokines and leukocyte infiltration in brain and NF-κB signaling were evaluated after ischemia/reperfusion.

Results
MCPIP1 protein and mRNA levels significantly increased specifically in mouse brain undergoing EA pretreatment. EA pretreatment significantly attenuated the infarct volume, neurological deficits, upregulation of proinflammatory cytokines and leukocyte infiltration in the brain of wild-type mice after MCAO compared with that of the non-EA group. MCPIP1-deficient mice failed to evoke EA pretreatment-induced tolerance compared with that of the control MCPIP1 knockout group without EA treatment. Furthermore, the activation of NF-κB signaling was significantly reduced in EA-pretreated wild-type mice after MCAO compared to that of the non-EA control group and MCPIP1-deficient mice failed to confer the EA pretreatment-induced inhibition of NF-κB signaling after MCAO.

Conclusions
Our data demonstrated that MCPIP1 deficiency caused significant lack of EA pretreatment-induced cerebral protective effects after MCAO compared with the control group and that MCPIP1 is involved in EA pretreatment-induced delayed brain ischemia tolerance.
The Caenorhabditis elegans voltage-gated calcium channel subunits UNC-2 and UNC-36 and the calcium-dependent kinase UNC-43/CaMKII regulate neuromuscular junction morphology
Authors: Caylor Raymond C , Jin Yishi , Ackley Brian D 
Published: May 2013

Abstract

Background
The conserved Caenorhabditis elegans proteins NID-1/nidogen and PTP-3A/LAR-RPTP function to efficiently localize the presynaptic scaffold protein SYD-2/α-liprin at active zones. Loss of function in these molecules results in defects in the size, morphology and spacing of neuromuscular junctions.

Results
Here we show that the Cav2-like voltage-gated calcium channel (VGCC) proteins, UNC-2 and UNC-36, and the calmodulin kinase II (CaMKII), UNC-43, function to regulate the size and morphology of presynaptic domains in C. elegans. Loss of function in unc-2, unc-36 or unc-43 resulted in slightly larger GABAergic neuromuscular junctions (NMJs), but could suppress the synaptic morphology defects found in nid-1/ nidogen or ptp-3/ LAR mutants. A gain-of-function mutation in unc-43 caused defects similar to those found in nid-1 mutants. Mutations in egl-19, Cav1-like, or cca-1, Cav3-like, α1 subunits, or the second α2/δ subunit, tag-180, did not suppress nid-1, suggesting a specific interaction between unc-2 and the synaptic extracellular matrix (ECM) component nidogen. Using a synaptic vesicle marker in time-lapse microscopy studies, we observed GABAergic motor neurons adding NMJ-like structures during late larval development. The synaptic bouton addition appeared to form in at least two ways: (1) de novo formation, where a cluster of vesicles appeared to coalesce, or (2) when a single punctum became enlarged and then divided to form two discrete fluorescent puncta. In comparison to wild type animals, we found unc-2 mutants exhibited reduced NMJ dynamics, with fewer observed divisions during a similar stage of development.

Conclusions
We identified UNC-2/UNC-36 VGCCs and UNC-43/CaMKII as regulators of C. elegans synaptogenesis. UNC-2 has a modest role in synapse formation, but a broader role in regulating dynamic changes in the size and morphology of synapses that occur during organismal development. During the late 4th larval stage (L4), wild type animals exhibit synaptic morphologies that are similar to those found in animals lacking NID-1/PTP-3 adhesion, as well as those with constitutive activation of UNC-43. Genetic evidence indicates that the VGCCs and the NID-1/PTP-3 adhesion complex provide opposing functions in synaptic development, suggesting that modulation of synaptic adhesion may underlie synapse development in C. elegans.
A standard vector for the chromosomal integration and characterization of BioBrick™ parts in Escherichia coli
Authors: Zucca Susanna , Pasotti Lorenzo , Politi Nicolò , Cusella De Angelis Maria Gabriella , Magni Paolo 
Published: May 2013

Abstract

Background
The chromosomal integration of biological parts in the host genome enables the engineering of plasmid-free stable strains with single-copy insertions of the desired gene networks. Although different integrative vectors were proposed, no standard pre-assembled genetic tool is available to carry out this task. Synthetic biology concepts can contribute to the development of standardized and user friendly solutions to easily produce engineered strains and to rapidly characterize the desired genetic parts in single-copy context.

Results
In this work we report the design of a novel integrative vector that allows the genomic integration of biological parts compatible with the RFC10, RFC23 and RFC12 BioBrick™ standards in Escherichia coli. It can also be specialized by using BioBrick™ parts to target the desired integration site in the host genome. The usefulness of this vector has been demonstrated by integrating a set of BioBrick™ devices in two different loci of the E. coli chromosome and by characterizing their activity in single-copy. Construct stability has also been evaluated and compared with plasmid-borne solutions.

Conclusions
Physical modularity of biological parts has been successfully applied to construct a ready-to-engineer BioBrick™ vector, suitable for a stable chromosomal insertion of standard parts via the desired recombination method, i.e. the bacteriophage integration mechanism or homologous recombination. In contrast with previously proposed solutions, it is a pre-assembled vector containing properly-placed restriction sites for the direct transfer of various formats of BioBrick™ parts. This vector can facilitate the characterization of parts avoiding copy number artefacts and the construction of antibiotic resistance-free engineered microbes, suitable for industrial use.
In vitro evaluation of β-carboline alkaloids as potential anti-Toxoplasma agents
Authors: Alomar Maria L , Rasse-Suriani Federico AO , Ganuza Agustina , Cóceres Verónica M , Cabrerizo Franco M , Angel Sergio O 
Published: May 2013

Abstract

Background
Toxoplasmosis is a worldwide infection caused by the protozoan parasite Toxoplasma gondii, which causes chorioretinitis and neurological defects in congenitally infected newborns or immunodeficient patients. The efficacy of the current treatment is limited, primarily by serious host toxicity. In recent years, research has focused on the development of new drugs against T. gondii. β-Carbolines (βCs), such as harmane, norharmane and harmine, are a group of naturally occurring alkaloids that show microbicidal activity. In this work, harmane, norharmane and harmine were tested against T. gondii.

Findings
The treatment of extracellular tachyzoites with harmane, norharmane and harmine showed a 2.5 to 3.5-fold decrease in the invasion rates at doses of 40 μM (harmane and harmine) and 2.5 μM (norharmane) compared with the untreated parasites. Furthermore, an effect on the replication rate could also be observed with a decrease of 1 (harmane) and 2 (norharmane and harmine) division rounds at doses of 5 to 12.5 μM. In addition, the treated parasites presented either delayed or no monolayer lysis compared with the untreated parasites.

Conclusions
The three βC alkaloids studied (norharmane, harmane and harmine) exhibit anti-T. gondii effects as evidenced by the partial inhibition of parasite invasion and replication. A dose–response effect was observed at a relatively low drug concentration (< 40 μM), at which no cytotoxic effect was observed on the host cell line (Vero).
Accuracy of tympanic temperature measurement using an infrared tympanic membrane thermometer
Authors: Gasim Gasim I , Musa Imad R , Abdien Mohamed T , Adam Ishag 
Published: May 2013

Abstract

Background
During investigation and diagnosis of patients, accurate temperature measurement is of great importance. The advantages of tympanic membrane thermometry are speed (temperature reading available within seconds), safety, and ease of use. The aim of this study was to compare the accuracy of infrared tympanic thermometers in comparison to mercury thermometers in measurement of body temperature.

Methods
Axillary and tympanic temperature was measured simultaneously in consecutive patients using mercury glass and infrared tympanic thermometers at Omdurman Hospital, Sudan during October 2012.

Results
In total, temperature was measured in 174 patients, 95 of whom (54.6%) were male. The mean (SD) patient age and weight was 33.18 (25.07) years and 52.13 (69.85) kg. There was no significant difference in mean (SD) temperature measurement between mercury and infrared tympanic membrane thermometers, 37.29°C (0.91) versus 37.38°C (0.95), P = 0.373, respectively. There was a significant positive correlation between axillary and tympanic body temperature measurements (r = 0.697, P < 0.001). The mean difference between the two readings (with limits of agreements) was - 0.093 (−0.20; 0.02) °C.

Conclusion
In this study, tympanic membrane thermometry is as reliable and accurate as axillary mercury glass thermometry. Thus, tympanic thermometry can be used in clinical practice, especially in the emergency setting, where ease of use and speed of obtaining the temperature reading are important.
A randomized trial of video self-instruction in cardiopulmonary resuscitation for lay persons
Authors: Godfred Rachel , Huszti Ella , Fly Deborah , Nichol Graham 
Published: May 2013

Abstract

Background
Cardiopulmonary resuscitation (CPR) improves outcomes after cardiac arrest. Much of the lay public is untrained in CPR skills. We evaluated the effectiveness of a compression-only CPR video self-instruction (VSI) with a personal manikin in the lay public.

Methods
Adults without prior CPR training in the past year or responsibility to provide medical care were randomized into one of three groups: 1) Untrained before testing, 2) 10-minute VSI in compressions-only CPR (CPR Anytime, American Heart Association, Dallas, TX), or 3) 22-minute VSI in compressions and ventilations (CPR Anytime). CPR proficiency was assessed using a sensored manikin. The primary outcome was composite skill competence of 90% during five minutes of skill demonstration. Evaluated were alternative cut-points for skill competence and individual components of CPR. 488 subjects (143 in untrained group, 202 in compressions-only group and 143 in compressions and ventilation group) were required to detect 21% competency with compressions-only versus 7% with untrained and 34% with compressions and ventilations.

Results
Analyzable data were available for the untrained group (n = 135), compressions-only group (n = 185) and the compressions and ventilation group (n = 119). Four (3%) achieved competency in the untrained group (p-value = 0.57 versus compressions-only), nine (4.9%) in the compressions-only group, and 12 (10.1%) in the compressions and ventilations group (p-value 0.13 vs. compressions-only). The compressions-only group had a greater proportion of correct compressions (p-value = 0.028) and compressions with correct hand placement (p-value = 0.0004) compared to the untrained group.

Conclusions
VSI in compressions-only CPR did not achieve greater overall competency but did achieve some CPR skills better than without training.
Rapid, responsive, relevant (R3) research: a call for a rapid learning health research enterprise
Authors: Riley William T , Glasgow Russell E , Etheredge Lynn , Abernethy Amy P 
Published: May 2013

AbstractOur current health research enterprise is painstakingly slow and cumbersome, and its results seldom translate into practice. The slow pace of health research contributes to findings that are less relevant and potentially even obsolete. To produce more rapid, responsive, and relevant research, we propose approaches that increase relevance via greater stakeholder involvement, speed research via innovative designs, streamline review processes, and create and/or better leverage research infrastructure. Broad stakeholder input integrated throughout the research process can both increase relevance and facilitate study procedures. More flexible and rapid research designs should be considered before defaulting to the traditional two‐arm randomized controlled trial (RCT), but even traditional RCTs can be designed for more rapid findings. Review processes for grant applications, IRB protocols, and manuscript submissions can be better streamlined to minimize delays. Research infrastructures such as rapid learning systems and other health information technologies can be leveraged to rapidly evaluate new and existing treatments, and alleviate the extensive recruitment delays common in traditional research. These and other approaches are feasible but require a culture shift among the research community to value not only methodological rigor, but also the pace and relevance of research.
Generation of gene disruptions by transcription activator-like effector nucleases (TALENs) in Xenopus tropicalis embryos
Authors: Lei Yong , Guo Xiaogang , Deng Yi , Chen Yonglong , Zhao Hui 
Published: May 2013

Abstract
Transcription activator-like effector nucleases (TALENs) are novel engineered DNA nucleases, and have been proven to be effective for gene specific targeting in various species. Recently we reported gene disruptions in Xenopus embryos by using TALENs. Here we summarize the protocol that is used in our studies for gene disruption. This protocol covers selection of TALEN targeting sites, TALEN assembly with a modified Golden Gate method, and injection of TALEN mRNAs into Xenopus tropicalis embryos. We also provide details for detection of somatic and germ line transmitted mutations. And finally, we briefly describe establishment of knockout Xenopus lines. This protocol will facilitate broader applications of TALENs in studies of Xenopus biology.