bluuebunny/crossref_metadata_2025 huggingface dataset:
Variability of Water Chemistry in Tundra Lakes, Petuniabukta Coast, Central Spitsbergen, Svalbard
Authors: Mazurek Małgorzata , Paluszkiewicz Renata , Rachlewicz Grzegorz , Zwoliński Zbigniew
Published: May 2012

Samples of water from small tundra lakes located on raised marine terraces on the eastern coast of Petuniabukta (Ebbadalen, Central Spitsbergen) were examined to assess the changes in water chemistry that had occurred during the summer seasons of 2001–2003 and 2006. The unique environmental conditions of the study region include the predominance of sedimentary carbonate and sulphate rocks, low precipitation values, and an active permafrost layer with a maximum thickness of 1.2 m. The average specific electric conductivity (EC) values for the three summer seasons in the four lakes ranged from 242 to 398 μS cm−1. The highest EC values were observed when the air temperature decreased and an ice cover formed (cryochemical effects). The ion composition was dominated by calcium (50.7 to 86.6%), bicarbonates (39.5 to 86.4%), and sulphate anions. The high concentrations of , , and Ca2+ions were attributed to the composition of the bedrock, which mainly consists of gypsum and anhydrite. The average proportion of marine components in the total load found in the Ebbadalen tundra lake waters was estimated to be 8.1%. Precipitation supplies sulphates (as much as 69–81%) and chlorides (14–36%) of nonsea origin. The chief source of these compounds may be contamination from the town of Longyearbyen. Most ions originate in the crust, the active layer of permafrost, but some are atmospheric in origin and are either transported or generated in biochemical processes. The concentrations of most components tend to increase during the summer months, reaching a maximum during freezing and partially precipitating onto the bottom sediments.
Lethal Effects ofAspergillus nigeragainst Mosquitoes Vector of Filaria, Malaria, and Dengue: A Liquid Mycoadulticide
Authors: Singh Gavendra , Prakash Soam
Published: May 2012

Aspergillus nigeris a fungus of the genusAspergillus. It has caused a disease called black mold on certain fruits and vegetables. The culture filtrates released from theA. nigerATCC 66566 were grown in Czapek dox broth (CDB) then filtered with flash chromatograph and were used for the bioassay after a growth of thirty days. The result demonstrated these mortalities with LC50, LC90, and LC99values ofCulex quinquefasciatus0.76, 3.06, and 4.75,Anopheles stephensi1.43, 3.2, and 3.86, andAedes aegypti1.43, 2.2, and 4.1 μl/cm2, after exposure of seven hours. We have calculated significant LT90values ofCx. quinquefasciatus4.5,An. stephensi3.54, andAe. aegypti6.0 hrs, respectively. This liquid spray of fungal culture isolate ofA. nigercan reduce malaria, dengue, and filarial transmission. These results significantly support broadening the current vector control paradigm beyond chemical adulticides.
Are Fish and Standardized FETAX Assays Protective Enough for Amphibians? A Case Study onXenopus laevisLarvae Assay with Biologically Active Substances Present in Livestock Wastes
Authors: Martini Federica , Tarazona José V. , Pablos M. Victoria
Published: May 2012

Biologically active substances could reach the aquatic compartment when livestock wastes are considered for recycling. Recently, the standardized FETAX assay has been questioned, and some researchers have considered that the risk assessment performed on fish could not be protective enough to cover amphibians. In the present study aXenopus laevisacute assay was developed in order to compare the sensitivity of larvae relative to fish or FETAX assays; veterinary medicines (ivermectin, oxytetracycline, tetracycline, sulfamethoxazole, and trimethoprim) and essential metals (zinc, copper, manganese, and selenium) that may be found in livestock wastes were used for the larvae exposure. Lethal (LC50) and sublethal effects were estimated. Available data in both, fish and FETAX studies, were in general more protective than values found out in the current study, but not in all cases. Moreover, the presence of nonlethal effects, caused by ivermectin, zinc, and copper, suggested that several physiological mechanisms could be affected. Thus, this kind of effects should be deeply investigated. The results obtained in the present study could expand the information about micropollutants from livestock wastes on amphibians.
Benchmarking Strategies for Measuring the Quality of Healthcare: Problems and Prospects
Authors: Lovaglio Pietro Giorgio
Published: May 2012

Over the last few years, increasing attention has been directed toward the problems inherent to measuring the quality of healthcare and implementing benchmarking strategies. Besides offering accreditation and certification processes, recent approaches measure the performance of healthcare institutions in order to evaluate their effectiveness, defined as the capacity to provide treatment that modifies and improves the patient’s state of health. This paper, dealing with hospital effectiveness, focuses on research methods for effectiveness analyses within a strategy comparing different healthcare institutions. The paper, after having introduced readers to the principle debates on benchmarking strategies, which depend on the perspective and type of indicators used, focuses on the methodological problems related to performing consistent benchmarking analyses. Particularly, statistical methods suitable for controlling case-mix, analyzing aggregate data, rare events, and continuous outcomes measured with error are examined. Specific challenges of benchmarking strategies, such as the risk of risk adjustment (case-mix fallacy, underreporting, risk of comparing noncomparable hospitals), selection bias, and possible strategies for the development of consistent benchmarking analyses, are discussed. Finally, to demonstrate the feasibility of the illustrated benchmarking strategies, an application focused on determining regional benchmarks for patient satisfaction (using 2009 Lombardy Region Patient Satisfaction Questionnaire) is proposed.
Ultraviolet-B Radiation and Nitrogen Affect Nutrient Concentrations and the Amount of Nutrients Acquired by Above-Ground Organs of Maize
Authors: Correia Carlos M. , Coutinho João F. , Bacelar Eunice A. , Gonçalves Berta M. , Björn Lars Olof , Moutinho Pereira José
Published: May 2012

UV-B radiation effects on nutrient concentrations in above-ground organs of maize were investigated at silking and maturity at different levels of applied nitrogen under field conditions. The experiment simulated a 20% stratospheric ozone depletion over Portugal. At silking, UV-B increased N, K, Ca, and Zn concentrations, whereas at maturity Ca, Mg, Zn, and Cu increased and N, P and Mn decreased in some plant organs. Generally, at maturity, N, Ca, Cu, and Mn were lower, while P, K, and Zn concentrations in stems and nitrogen-use efficiency (NUE) were higher in N-starved plants. UV-B and N effects on shoot dry biomass were more pronounced than on nutrient concentrations. Nutrient uptake decreased under high UV-B and increased with increasing N application, mainly at maturity harvest. Significant interactions UV-B x N were observed for NUE and for concentration and mass of some elements. For instance, under enhanced UV-B, N, Cu, Zn, and Mn concentrations decreased in leaves, except on N-stressed plants, whereas they were less affected by N nutrition. In order to minimize nutritional, economical, and environmental negative consequences, fertiliser recommendations based on element concentration or yield goals may need to be adjusted.
Surgical Treatment of Patients with Lennox-Gastaut Syndrome Phenotype
Authors: Liu Shi-Yong , An Ning , Fang Xiang , Singh Prabhdeep , Oommen Joseph , Yin Qing , Yang Mei-Hua , Liu Yong , Liao Wei , Gao Chang-Qing , Yang Hui
Published: May 2012

Lennox-Gastaut syndrome (LGS) is a devastating and refractory generalized epilepsy affecting children and adolescents. In this study we report the results of resective surgery in 18 patients with LGS phenotype who underwent single-lobe/lesionectomy or multilobe resection plus multiple subpial transection and/or callosotomy. After surgery, seven patients became completely seizure-free (Engel Class I) and five almost seizure-free (Engel Class II). Additional four had significant seizure control (Engel Class III), and two had no change in seizure frequency (Engel Class IV). Of the 4 patients without any lesion on brain MRI, 2 ended with Engel Class II, 1 with III and the other with IV in Engels’ classification. Mean intelligence quotient (IQ) increased from 56.1 ± 8.1 (mean ± SD) before operation to 67.4 ± 8.2 (mean ± SD) after operation, a significant improvement (P=0.001). Results also indicated that the younger the patient at surgery, or the shorter the interval between onset of seizure and resective operation, the better the intellectual outcome. Our data suggest that resective epilepsy surgery can be successful in patients with LGS phenotype as long as the EEG shows dominance of discharges in one hemisphere and corresponding ipsilateral imaging findings, even with contralateral ictal discharges.
Insecticidal Activity ofBacillus thuringiensisStrains Isolated from Soil and Water
Authors: Konecka Edyta , Baranek Jakub , Hrycak Anita , Kaznowski Adam
Published: May 2012

We attempted to search novelBacillus thuringiensisstrains that produce crystals with potential utility in plant protection and with higher activity than strains already used in biopesticide production. SevenB. thuringiensissoil and water isolates were used in the research. We predicted the toxicity of their crystals bycrygene identification employing PCR method. The isolate MPU B63 with interesting, according to us, genes content was used in evaluating its crystal toxicity againstCydia pomonellacaterpillars. The strain MPU B63 was cultured from water sample and hadcry1Ab,cry1B, andcry15genes. The LC50crystals of MPU B63 were compared to LC50of commercial bioinsecticide Foray determined againstC. pomonella(codling moth). The activity of MPU B63 inclusions against codling moth larvae was approximately 24-fold higher than that of Foray. The results are a promising introduction for further study evaluating the potential usefulness of isolate MPU B63 crystals in plant protection.
Relation of Secondhand Smoking to Mild Cognitive Impairment in Older Inpatients
Authors: Orsitto Giuseppe , Turi Vincenzo , Venezia Amedeo , Fulvio Francesco , Manca Cosimo
Published: May 2012

Up to now, controversy still exists regarding the role of secondhand smoking (SHS) in developing cognitive impairment. This study aimed to evaluate the prevalence of SHS in hospitalized older patients with cognitive deficit, particularly in those with mild cognitive impairment (MCI). Smoking history was classified into four groups: never smokers, former-active smokers/no SHS, active smokers, and secondhand smokers, and cognitive function into three levels: normal cognition (C), MCI, and dementia. A total of 933 older subjects with diagnoses of MCI (n=98), dementia (n=124), or C (n=711) were enrolled in this cross-sectional study. As expected, patients with dementia had significantly higher frequency of former-active smokers than cognitively normal. Moreover, patients with MCI showed a significantly higher frequency of active and secondhand smokers than patients with dementia or C. A smoking history is very frequent in older patients with dementia. Patients with MCI had even higher rate of exposure to active or secondhand smoking.
The Solid Phase Extraction of Some Metal Ions Using Palladium Nanoparticles Attached to Silica Gel Chemically Bonded by Silica-Bonded N-Propylmorpholine as New Sorbent prior to Their Determination by Flame Atomic Absorption Spectroscopy
Authors: Ghaedi M. , Rezakhani M. , Khodadoust S. , Niknam K. , Soylak M.
Published: May 2012

In this research at first palladium nanoparticle attached to a new chemically bonded silica gel has been synthesized and has been characterized with different techniques such as X-ray diffraction (XRD), fourier transform infrared (FT-IR), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). Then, this new sorbent (chemically modified silica gel with N-propylmorpholine (PNP-SBNPM)) was efficiently used for preconcentration of some metal ions in various food samples. The influence of effective variables including mass of sorbent, flow rate, pH of sample solutions and condition of eluent such as volume, type and concentration on the recoveries of understudy metal ions were investigated. Following the optimization of variables, the interfering effects of some foreign ions on the preconcentration and determination of the investigated metal ions described. At optimum values of variables, all investigated metal ions were efficiently recovered with efficiency more than 95%, relative standard deviation (RSD) between 2.4 and 2.8, and detection limit in the range of 1.4–2.7 ng mL−1. The present method is simple and rapidly applicable for the determination of the understudied metal ions (ng mL−1) in different natural food samples.
Identification of Proteins with Potential Osteogenic Activity Present in the Water-Soluble Matrix Proteins fromCrassostrea gigasNacre Using a Proteomic Approach
Authors: Oliveira Daniel V. , Silva Tomé S. , Cordeiro Odete D. , Cavaco Sofia I. , Simes Dina C.
Published: May 2012

Nacre, when implantedin vivoin bones of dogs, sheep, mice, and humans, induces a biological response that includes integration and osteogenic activity on the host tissue that seems to be activated by a set of proteins present in the nacre water-soluble matrix (WSM). We describe here an experimental approach that can accurately identify the proteins present in the WSM of shell mollusk nacre. Four proteins (three gigasin-2 isoforms and a cystatin A2) were for the first time identified in WSM ofCrassostrea gigasnacre using 2DE and LC-MS/MS for protein identification. These proteins are thought to be involved in bone remodeling processes and could be responsible for the biocompatibility shown between bone and nacre grafts. These results represent a contribution to the study of shell biomineralization process and opens new perspectives for the development of new nacre biomaterials for orthopedic applications.
Aerobic Fitness Evaluation during Walking Tests Identifies the Maximal Lactate Steady State
Authors: Puga Guilherme Morais , Kokubun Eduardo , Simões Herbert Gustavo , Nakamura Fabio Yuzo , Campbell Carmen Sílvia Grubert
Published: May 2012

Objective. The aim of this study was to verify the possibility of lactate minimum (LM) determination during a walking test and the validity of such LM protocol on predicting the maximal lactate steady-state (MLSS) intensity.Design. Eleven healthy subjects ( yr;  kg;  cm) performed LM tests on a treadmill, consisting of walking at 5.5  and with 20–22% of inclination until voluntary exhaustion to induce metabolic acidosis. After 7 minutes of recovery the participants performed an incremental test starting at 7% incline with increments of 2% at each 3 minutes until exhaustion. A polynomial modeling approach (LMp) and a visual inspection (LMv) were used to identify the LM as the exercise intensity associated to the lowest [bLac] during the test. Participants also underwent to 2–4 constant intensity tests of 30 minutes to determine the MLSS intensity.Results. There were no differences among LMv (%), LMp (%), and MLSS (%) and the Bland and Altman plots evidenced acceptable agreement between them.Conclusion. It was possible to identify the LM during walking tests with intensity imposed by treadmill inclination, and it seemed to be valid on identifying the exercise intensity associated to the MLSS.
Recognition for Positive Behavior as a Critical Youth Development Construct: Conceptual Bases and Implications on Youth Service Development
Authors: Law Ben M. F. , Siu Andrew M. H. , Shek Daniel T. L.
Published: May 2012

Recognition for positive behavior is an appropriate response of the social environment to elicit desirable external behavior among the youth. Such positive responses, rendered from various social systems, include tangible and intangible reinforcements. The following theories are used to explain the importance of recognizing positive behavior: operational conditioning, observational learning, self-determination, and humanistic perspective. In the current work, culturally and socially desirable behaviors are discussed in detail with reference to Chinese adolescents. Positive behavior recognition is especially important to adolescent development because it promotes identity formation as well as cultivates moral reasoning and social perspective thinking from various social systems. The significance of recognizing positive behavior is illustrated through the support, tutorage, invitation, and subsidy provided by Hong Kong’s social systems in recognition of adolescent volunteerism. The practical implications of positive behavior recognition on youth development programs are also discussed in this work.
Routine Pulmonary Function Test Can Estimate the Extent of Tuberculous Destroyed Lung
Authors: Lee Eun Joo , Lee Sang Yeub , In Kwang Ho , Yoo Se Hwa , Choi Eun Jeong , Oh Yu Whan , Park Sanghoon
Published: May 2012

Tuberculous destroyed lung (TDL) is diagnosed by a clear past history of tuberculosis with findings of parenchymal destruction verified by chest X-ray. Despite the resultant deterioration of lung function and quality of lives seen in TDL patients, the exact mechanism or characteristics of pulmonary function worsening have not been clearly studied. We investigated the feature of respiratory impairment of TDL patients, and studied whether extent of destroyed lung measured with chest CT has any correlation with routine lung function. To evaluate the degree of destruction, the Goddard classification scoring system was modified into a novel scoring system (destroyed lung score, (DLS)) with a score from 0 to 4. Twenty-five subjects were enrolled. TDL predominantly manifested as an obstructive pattern (64%, 16/25). Median value of DLS of the entire lung was 2.6 (1.7–3.9). Absolute values of FEV1 and FVC were both negatively associated with DLS (r=-0.78,P=0.001, andr=-0.61,P=0.021). Percentage of predicted value of FEV1and FVC were also negatively associated with DLS (r=-0.62,P=0.019, andr=-0.76,P=0.002). Our study shows that lung function of TDL patients were notably correlated with the extent of destroyed lung measured with chest CT scan.
Trace Level Arsenic Quantification through Cloud Point Extraction: Application to Biological and Environmental Samples
Authors: Suresh Kumar Kempahanumakkagari , Pandurangappa Malingappa
Published: May 2012

A sensitive solvent-free extraction protocol for the quantification of arsenic at trace level has been described. It is based on the reaction of arsenic (V) with molybdate in acidic medium in presence of antimony (III) and ascorbic acid as a reducing agent to form a blue-colored arsenomolybdenum blue complex. The complex has been extracted into surfactant phase using Triton X-114, and its absorbance was measured at 690 nm. The detection limit, working range, and the relative standard deviation were found to be 1 ng mL−1, 10–200 ng mL−1, and 1.2%, respectively. The effect of common ions was studied, and the method has been applied to determine trace levels of As(III) and As(V) from a variety of samples like environmental, biological, and commercially procured chemicals.
Rodent Damage to Natural and Replanted Mountain Forest Regeneration
Authors: Heroldová Marta , Bryja Josef , Jánová Eva , Suchomel Josef , Homolka Miloslav
Published: May 2012

Impact of small rodents on mountain forest regeneration was studied in National Nature Reserve in the Beskydy Mountains (Czech Republic). A considerable amount of bark damage was found on young trees (20%) in spring after the peak abundance of field voles (Microtus agrestis) in combination with long winter with heavy snowfall. In contrast, little damage to young trees was noted under high densities of bank voles (Myodes glareolus) with a lower snow cover the following winter. The bark of deciduous trees was more attractive to voles (22% damaged) than conifers (8%). Young trees growing in open and grassy localities suffered more damage from voles than those under canopy of forest stands (, ). Natural regeneration in Nature Reserve was less damaged compared to planted trees (, ). The main factors influencing the impact of rodent species on tree regeneration were open, grassy habitat conditions, higher abundance of vole species, tree species preferences- and snow-cover condition. Under these conditions, the impact of rodents on forest regeneration can be predicted. Foresters should prefer natural regeneration to the artificial plantings.
Purification and Characterization of Hemagglutinating Proteins from Poker-Chip Venus (Meretrix lusoria) and Corbicula Clam (Corbicula fluminea)
Authors: Cheng Chin-Fu , Hung Shao-Wen , Chang Yung-Chung , Chen Ming-Hui , Chang Chen-Hsuan , Tsou Li-Tse , Tu Ching-Yu , Lin Yu-Hsing , Liu Pan-Chen , Lin Shiun-Long , Wang Way-Shyan
Published: May 2012

Hemagglutinating proteins (HAPs) were purified from Poker-chip Venus (Meretrix lusoria) and Corbicula clam (Corbicula fluminea) using gel-filtration chromatography on a Sephacryl S-300 column. The molecular weights of the HAPs obtained from Poker-chip Venus and Corbicula clam were 358 kDa and 380 kDa, respectively. Purified HAP from Poker-chip Venus yielded two subunits with molecular weights of 26 kDa and 29 kDa. However, only one HAP subunit was purified from Corbicula clam, and its molecular weight was 32 kDa. The two Poker-chip Venus HAPs possessed hemagglutinating ability (HAA) for erythrocytes of some vertebrate animal species, especially tilapia. Moreover, HAA of the HAP purified from Poker-chip Venus was higher than that of the HAP of Corbicula clam. Furthermore, Poker-chip Venus HAPs possessed better HAA at a pH higher than 7.0. When the temperature was at 4°C–10°C or the salinity was less than 0.5‰, the two Poker-chip Venus HAPs possessed better HAA compared with that of Corbicula clam.
Program Implementers' Evaluation of the Project P.A.T.H.S.: Findings Based on Different Datasets over Time
Authors: Shek Daniel T. L. , Ma Cecilia M. S.
Published: May 2012

This paper integrates the evaluation findings based on program implementers in nine datasets collected from 2005 to 2009 (244 schools and 7,926 implementers). Using consolidated data with schools as the unit of analysis, results showed that program implementers generally had positive perceptions of the program, themselves, and benefits of the program, with more than four-fifths of the implementers regarding the program as beneficial to the program participants. The subjective outcome evaluation instrument was found to be internally consistent. Multiple regression analyses revealed that perceived qualities of the program and program implementers predicted perceived effectiveness of the program. In conjunction with evaluation findings based on other sources, the present study provides support for the effectiveness of the Tier 1 Program of the Project P.A.T.H.S. (Positive Adolescent Training through Holistic Social Programmes) in Hong Kong.
Photoinduced Synthesis of New Diisochromenochromen-4-ones and Their Antimicrobial Activities
Authors: Yusuf Mohamad , Solanki Indu , Jain Payal
Published: May 2012

The diisochromenochromen-4-one3a-3b,4a-4c,5a-6a  & 7have been prepared from the photocyclization reaction of bischromen-4-one2a-2e. The later compounds are obtained from the O-alkylation of the suitable 3-hydroxy-2-aryl-4H-chromen-4-one1a-1ewith 4,4′-bischloromethyl-diphenyl in dry acetone, anhydrous K2CO3, and PTC (Bu4N+I−) under refluxing conditions. The structures of compounds2a-2e,3a-3b,4a-4c,5a-6a  & 7have been characterized from the rigorous analysis of their IR,1H-NMR,13C-NMR, ESI-Mass, and elemental analysis. The antibacterial and antifungal activities of the synthesized products were also evaluated against theKlebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Bacillus subtilis, andAspergillus janusandPenicillium glabrum, respectively. Some of the tested compounds showed significant activity against the above-said microorganisms.
Codependency of H2B monoubiquitination and nucleosome reassembly on Chd1
Authors: Lee Jung-Shin , Garrett Alexander S. , Yen Kuangyu , Takahashi Yoh-Hei , Hu Deqing , Jackson Jessica , Seidel Christopher , Pugh B. Franklin , Shilatifard Ali
Published: May 2012

Monoubiquitination of histone H2B on Lys 123 (H2BK123ub) is a transient histone modification considered to be essential for establishing H3K4 and H3K79 trimethylation by Set1/COMPASS and Dot1, respectively. Here, we identified Chd1 as a factor that is required for the maintenance of high levels of H2B monoubiquitination, but not for H3K4 and H3K79 trimethylation. Loss of Chd1 results in a substantial loss of H2BK123ub levels with little to no effect on the genome-wide pattern of H3K4 and H3K79 trimethylation. Our data show that nucleosomal occupancy is reduced in gene bodies in both chd1Δ and, as has been shown, K123A mutant backgrounds. We also demonstrated that Chd1's function in maintaining H2BK123ub levels is conserved from yeast to humans. Our study provides evidence that only small levels of H2BK123ub are necessary for full levels of H3K4 and H3K79 trimethylation in vivo and points to a possible role for Chd1 in positively regulating gene expression through promoting nucleosome reassembly coupled with H2B monoubiquitination.
Formaldehyde Fixation of Drosophila Tissues onto Slides for Whole-Mount FISH
Authors: Dernburg Abby F.
Published: May 2012

This protocol describes the formaldehyde fixation of Drosophila tissues other than egg chambers. It provides a general approach to fixing tissue or cells onto microscope slides in a manner compatible with whole-mount fluorescent in situ hybridization (FISH) analysis. The procedure includes a simple formaldehyde fixation in buffer, followed by postfixation in cold ethanol. Postfixation of the tissue in methanol or ethanol can markedly improve permeability. In some instances, it may be preferable to fix tissue onto coverslips, because microscope optics are usually designed to optimize imaging immediately adjacent to the coverslip. On the other hand, coverslips are much more fragile and harder to manipulate.
Site-Specific Integration with Bacteriophage ɸC31 Integrase
Authors: Hillman R. Tyler , Calos Michele P.
Published: May 2012

Few nonviral techniques exist for efficient and stable eukaryotic gene transfer and fewer still are broadly useful in both cell culture and whole-organism applications. ɸC31 integrase, a site-specific bacteriophage recombinase, is able to catalyze chromosomal transgene insertion under a diverse range of experimental and therapeutic conditions. The enzyme recognizes and catalyzes unidirectional recombination between attachment motifs found in phage and bacterial genomes (attP and attB sites, respectively). Use of ɸC31 integrase for gene transfer requires that an attB sequence be cloned into a transgene-bearing plasmid. When this modified plasmid is introduced into cells alongside integrase-expressing plasmid, ɸC31 integrase is able to catalyze insertion of the transgene plasmid into one of a limited pool of sites in the target genome that show sequence similarity to wild-type attP. Efficient delivery of ɸC31 integrase and attB donor plasmid to the tissue or cells of interest remains the most challenging aspect of the system. Unlike viral methods of genome manipulation, use of ɸC31 integrase almost always requires an additional method of stimulating cellular DNA uptake. However, the relative simplicity of the plasmid-based system means that nearly any proven method of introducing exogenous DNA into cells can be used with ɸC31 integrase. This protocol describes the use of ɸC31 integrase in mammalian cell culture for the creation of clonal lines showing robust and stable expression of an experimental transgene.
Preparing Drosophila Larvae for Feeding Assays
Authors: Shen Ping
Published: May 2012

The food responses of Drosophila larvae offer an excellent opportunity to study the genetic and neural regulation of feeding behavior. Compared with fed larvae, hungry larvae are more likely to display aggressive foraging, rapid food intake, compensatory feeding, and stress-resistant food procurement. Behavioral assays have been developed to quantitatively assess particular aspects of the hunger-driven food response. In combination, these assays help define the specific role of signaling molecules or neurons in the regulation of feeding behavior in foraging larvae. Proper rearing of larvae is of paramount importance for all behavioral experiments. A time-tested procedure is described here to produce highly synchronized third-instar feeding larvae. To ensure synchronized growth of larvae, the rearing conditions must be optimized by controlling food quality, larval density, humidity, and temperature. Under such conditions, it takes ∼ 72 h for a wild-type or white egg to develop into a third-instar larva. One reliable way to judge the age of third-instar larvae is to determine the time of the second molting (into the third instar). A feeding step is included to ensure the selection of young third instars that are actively foraging to use in behavioral assays.
Analysis of Feeding Behavior of Drosophila Larvae on Liquid Food
Authors: Shen Ping
Published: May 2012

The food responses of Drosophila larvae offer an excellent opportunity to study the genetic and neural regulation of feeding behavior. Compared with fed larvae, hungry larvae are more likely to display aggressive foraging, rapid food intake, compensatory feeding, and stress-resistant food procurement. Behavioral assays have been developed to quantitatively assess particular aspects of the hunger-driven food response. In combination, these assays help define the specific role of signaling molecules or neurons in the regulation of feeding behavior in foraging larvae. This protocol describes the analysis of larvae feeding on liquid food. The test is designed for quantitative assessment of the food ingestion rate of individual larvae under different energy states. It provides a simple and reliable way to measure the graded modification of the baseline feeding rate of larvae as food deprivation is prolonged. The test is applicable to routine functional testing and larger-scale screening of genetic mutations and biologics that might affect food consumption.
Fluorescent Labeling of Myosin V for Polarized Total Internal Reflection Fluorescence Microscopy (polTIRFM) Motility Assays
Authors: Beausang John F. , Sun Yujie , Quinlan Margot E. , Forkey Joseph N. , Goldman Yale E.
Published: May 2012

Polarized total internal reflection fluorescence microscopy (polTIRFM) can be used to detect the spatial orientation and rotational dynamics of single molecules. polTIRFM determines the three-dimensional angular orientation and the extent of wobble of a fluorescent probe bound to the macromolecule of interest. This protocol describes how to exchange bifunctional rhodamine–calmodulin (BR-CaM) for wild-type calmodulin (WT-CaM) on the lever arm of myosin V. BR-CaM is exchanged at low stoichiometry (∼0.4 BR-CaM per double-headed myosin V) to obtain myosin V molecules with one BR-CaM and to limit the proportion of myosin V molecules with two or more probes. The stoichiometry is very sensitive to the concentration of calcium during the exchange reaction. The labeled myosin V can subsequently be used for investigating the motility of myosin V in vitro with a polTIRFM processive motility assay, which is performed on substrate-attached actin.
17βH-Periplogenin, a cardiac aglycone from the root bark of Periploca sepium Bunge
Authors: Zhang Yu-Wei , Bao Yong-Li , Wu Yin , Yu Chun-Lei , Li Yu-Xin
Published: May 2012

The title compound {systematic name: 4-[(3S,5S,8R,9S,10R,13R,14S,17S)-3,5,14-trihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl]furan-2(5H)-one}, C23H34O5, was isolated from the roots of Periploca sepium Bunge, a famous Chinese traditional herbal medicine. The three six-membered rings adopt chair conformations, the cyclopentane ring displays an approximate envelope conformation (with the C atom bearing the methyl substituent at the flap) and the five-membered lactone ring adopts an essentially planar [maximum deviation of 0.004 (8) Å] conformation. In the crystal, molecules are linked into helical chains along [010] by O—H...O hydrogen bonds and weak C—H...O interactions. Two intramolecular O—H...O hydrogen bonds are also present.
2,6-Dichloro-7-isopropyl-7H-purine
Authors: Hloušková Nikola , Rouchal Michal , Nečas Marek , Vícha Robert
Published: May 2012

In the title molecule, C8H8Cl2N4, the essentially planar imidazole and pyrimidine rings [maximum deviations of 0.0030 (15) and 0.0111 (15) Å, respectively] make a dihedral angle of 1.32 (8)°. In the crystal, the fused-ring systems are stacked approximately parallel to the bc plane, with a centroid–centroid distance between inversion-related pyrimidine rings of 3.5189 (9) Å.
Dimethyl 2-[22,24-dimethyl-23-oxo-8,11,14-trioxa-25-azatetracyclo[19.3.1.02,7.015,20]pentacosa-2,4,6,15(20),16,18-hexaen-25-yl]but-2-enedioate
Authors: Anh Le Tuan , Hieu Truong Hong , Soldatenkov Anatoly T. , Kolyadina Nadezhda M. , Khrustalev Victor N.
Published: May 2012

The title compound, C29H33NO8, is a product of the Michael addition of the cyclic secondary amine subunit of the aza-14-crown-4 ether to dimethyl acetylenedicarboxylate. The piperidinone ring exhibits a distorted chair conformation, and the dimethyl ethylenedicarboxylate fragment has a cis configuration with a dihedral angle of 78.96 (5)° between the two carboxylate groups. The crystal packing is stabilized by weak C—H...O hydrogen bonds.
9-(2-Hydroxy-4,4-dimethyl-6-oxocyclohex-1-en-1-yl)-3,3-dimethyl-2,3,4,9-tetrahydro-1H-xanthen-1-one
Authors: Kurbanova Malahat M. , Gurbanov Atash V. , Mahmudov Kamran T. , Maharramov Abel M. , Ng Seik Weng
Published: May 2012

The cyclohexene ring that constitutes a part of the tetrahydroxanthene fused-ring system of the title compound, C23H26O4, adopts a flattened half-chair conformation that approximates an envelope conformation (in which the methylene C atom bearing the two methyl substituents represents the flap) as five of the six atoms lie approximately on a plane (r.m.s. deviation = 0.020 Å). The mean plane of the cyclohexene ring with the hydroxy substituent is approximately perpendicular to the mean plane of the tetrahydroxanthene system. In the crystal, adjacent molecules are linked by O—H...Ocarbonyl hydrogen bonds into a chain running along the b axis.
Effect of sub‐core scale heterogeneities on acoustic and electrical properties of a reservoir rock: a CO2 flooding experiment of brine saturated sandstone in a computed tomography scanner
Authors: Alemu Binyam L. , Aker Eyvind , Soldal Magnus , Johnsen Øistein , Aagaard Per
Published: May 2012

The effect of sub‐core scale heterogeneity on fluid distribution pattern, and the electrical and acoustic properties of a typical reservoir rock was studied by performing drainage and imbibition flooding tests with CO2 and brine in a laboratory. Moderately layered Rothbach sandstone was used as a test specimen. Two core samples were drilled; one perpendicular and the other parallel to the layering to allow injection of fluids along and normal to the bedding plane. During the test 3D images of fluid distribution and saturation levels were mapped by an industrial X‐ray CT‐scanner together with simultaneous measurement of electrical resistivity, ultrasonic velocities as well as amplitudes.

The results showed how the layering and the flooding direction influenced the fluid distribution pattern and the saturation level of the fluids. For a given fluid saturation level, the measured changes in the acoustic and electrical parameters were affected by both the fluid distribution pattern and the layering orientation relative to the measurement direction. The P‐wave amplitude and the electrical resistivity were more sensitive to small changes in the fluid distribution patterns than the P‐wave velocity. The change in amplitude was the most affected by the orientation of the layering and the resulting fluid distribution patterns. In some instances the change due to the fluid distribution pattern was higher than the variation caused by the change in CO2 saturation. As a result the Gassmann relation based on ‘uniform' or ‘patchy' saturation pattern was not suitable to predict the P‐wave velocity variation. Overall, the results demonstrate the importance of core‐imaging to improve our understanding of fluid distribution patterns and the associated effects on measured rock‐physics properties.
True amplitude elastic Gaussian beam imaging of multicomponent walkaway vertical seismic profiling data
Authors: Protasov M.I. , Tcheverda V.A.
Published: May 2012

We develop the true‐amplitude prestack migration of multicomponent data based on the use of elastic Gaussian beams for walkaway vertical seismic profile (VSP) acquisition systems. It consists in a weighted summation of multishot data with specific weights, computed by tracing elastic Gaussian beams from each imaging point of the target area towards the sources and receivers. Each pair of beams may be connected with either a pair of P‐rays (PP‐image) or the P‐ray towards sources and the S‐ray to receivers (PS‐image) and is uniquely determined by dip (the angle of the bisector between the rays and the vertical direction) and opening (the angle between the rays) angles. Shooting from the bottom towards the acquisition system helps to avoid well‐known troubles, in particular multipathing for the imaging conditions in complex velocity models. The ability to fix the dip angle and implement summation over opening angles leads to the so‐called selective images that contain mostly interfaces with desired slopes. On the other hand, a set of images computed for a range of opening angles by summation over all available dip angles is used as input of an AVO‐like inversion procedure for the recovery of elastic parameters.

The feasibility of this imaging procedure is verified by synthetic data for 2D realistic elastic models.
Transposon proliferation in an asexual parasitoid
Authors: KRAAIJEVELD KEN , ZWANENBURG BRECHTJE , HUBERT BENJAMIN , VIEIRA CRISTINA , De PATER SYLVIA , Van ALPHEN JACQUES J. M. , Den DUNNEN JOHAN T. , De KNIJFF PETER
Published: May 2012

The widespread occurrence of sex is one of the most elusive problems in evolutionary biology. Theory predicts that asexual lineages can be driven to extinction by uncontrolled proliferation of vertically transmitted transposable elements (TEs), which accumulate because of the inefficiency of purifying selection in the absence of sex and recombination. To test this prediction, we compared genome‐wide TE load between a sexual lineage of the parasitoid wasp Leptopilina clavipes and a lineage of the same species that is rendered asexual by Wolbachia‐induced parthenogenesis. We obtained draft genome sequences at 15–20× coverage of both the sexual and the asexual lineages using next‐generation sequencing. We identified transposons of most major classes in both lineages. Quantification of TE abundance using coverage depth showed that copy numbers in the asexual lineage exceeded those in the sexual lineage for DNA transposons, but not LTR and LINE‐like elements. However, one or a small number of gypsy‐like LTR elements exhibited a fourfold higher coverage in the asexual lineage. Quantitative PCR showed that high loads of this gypsy‐like TE were characteristic for 11 genetically distinct asexual wasp lineages when compared to sexual lineages. We found no evidence for an overall increase in copy number for all TE types in asexuals as predicted by theory. Instead, we suggest that the expansions of specific TEs are best explained as side effects of (epi)genetic manipulations of the host genome by Wolbachia. Asexuality is achieved in a myriad of ways in nature, many of which could similarly result in TE proliferation.
Role of Advanced Glycation Endproducts and Potential Therapeutic Interventions in Dialysis Patients
Authors: Mallipattu Sandeep K. , He John C. , Uribarri Jaime
Published: May 2012

It has been nearly 100 years since the first published report of advanced glycation end products (AGEs) by the French chemist Maillard. Since then, our understanding of AGEs in diseased states has dramatically changed. Especially in the last 25 years, AGEs have been implicated in complications related to aging, neurodegenerative diseases, diabetes, and chronic kidney disease. Although AGE formation has been well characterized by both in vitro and in vivo studies, few prospective human studies exist demonstrating the role of AGEs in patients on chronic renal replacement therapy. As the prevalence of end‐stage renal disease (ESRD) in the United States rises, it is essential to identify therapeutic strategies that either delay progression to ESRD or improve morbidity and mortality in this population. This article reviews the role of AGEs, especially those of dietary origin, in ESRD patients as well as potential therapeutic anti‐AGE strategies in this population.
The Effect of Early Postpartum Home Visits by Health Visitors: A Natural Experiment
Authors: Kronborg Hanne , Væth Michael , Kristensen Ingeborg
Published: May 2012

Objective

The purpose of this study was to assess if the absence of early home visits influenced the mothers’ breastfeeding duration and use of medical services.

Design

Data from mothers who had given birth during a strike period were compared with data from a reference period with usual work practice.

Sample

The study included 3,834 newborn and 375 health visitors, 75 of whom worked during the strike period.

Intervention

All families were offered nonstandardized home visits after discharge in the reference period. During the strike, the service was based on individual risk assessment.

Results

Overall, no difference in breastfeeding duration between the two periods was seen, but unvisited mothers in the strike period had shorter durations of full breastfeeding than a comparable group of mothers in the reference period (p < .005). Moreover, mothers in the strike period reported a significantly higher use of medical services. The mothers’ needs for postnatal visits differed depending on parity: primiparae underlined uncertainty, multiparae underlined previous breastfeeding experience. Mothers had missed out on guidance on all areas of the health visitors’ service.

Conclusion

Nonstandardized home visits by health visitors were associated with a longer breastfeeding duration. The postnatal visits depended on parity and unmet needs increased the use of medical services.
The Use of Asian Ficus Species for Restoring Tropical Forest Ecosystems
Authors: Kuaraksa Cherdsak , Elliott Stephen
Published: May 2012

Fig (Ficus spp.) trees have been promoted as framework species for tropical forest restoration throughout Asia, because they are considered to be keystone species. This article presents optimal propagation and planting techniques for six Asian dioecious Ficus species, which will enable their inclusion in forest restoration plantings across the Asia‐Pacific region: Ficus auriculata, F. fulva, F. hispida, F. oligodon, F. semicordata, and F. variegata. Nursery experiments compared the growth performance of propagating planting stock from seed and from leafy cuttings, whereas field experiments assessed the cost‐effectiveness and the relative performance of (1) direct seeding, (2) planting stock from seed, and (3) planting stock from cuttings. The most efficient method of producing Ficus spp. was from seed. Propagation from cuttings was much less successful. Seedlings produced from seed had the highest rates of growth and survival both in the nursery and in field trials. In field trials, use of planting stock from seed was also more cost‐effective than direct seeding and vegetative propagation. Establishment costs calculated on the basis of “per plant established” were $1.14 for seed, $6.95 for cutting, and $25.88 for direct seeding.
The phenology of molting, breeding and their overlap in central Amazonian birds
Authors: Johnson Erik I. , Stouffer Philip C. , Bierregaard, Jr Richard O.
Published: May 2012

The energetically challenging periods of molting and breeding are usually temporally separated in temperate birds, but can occur simultaneously in tropical birds, a condition known as molt–breeding overlap. Here, we document great variation in the timing and duration of molting and breeding, and in the extent of molt–breeding overlap, among 87 species of understory passerines in central Amazonia. We analyzed molt and breeding from 26 871 birds captured over a 30‐yr period near Manaus, Brazil. Although most species typically bred during the late dry season (about October through January), many thamnophilids apparently bred year‐round, whereas a few other species from a variety of families bred mainly during the wet season (about January through May). Of all breeding birds with an active brood patch, 12.7% were simultaneously molting. Molt–breeding overlap was more frequently observed among suboscines (13.3%), especially thamnophilids (23.0%), than oscines (6.4%). Some families had <5% molt–breeding overlap frequency, including Tyrannidae (4.4%), Tityridae (0.0%), Pipridae (1.5%), Turdidae (0.0%), and Thraupidae (0.0%), indicating that not all tropical species exhibit molt–breeding overlap. Among 31 well‐sampled species (n ≥15 brood patches), variation in molt–breeding overlap frequency was positively correlated with each species’ average duration of flight feather replacement (range 98–301 d). We also measured feather growth rates of individual birds in nine species; in five of these, slower‐growing feathers increased with an individual's probability of having molt–breeding overlap. Among furnariids, molt–breeding overlap occurred either at the beginning or end of the molt cycle, suggesting that physiological mechanisms typically separate molting from breeding. Thamnophilids showed a much different pattern; molt–breeding overlap occurred at any stage of feather replacement, apparently not regulated to be independent of breeding. These results reveal substantial life‐history variation among Amazonian birds. Future work to resolve the physiological regulation of molting and breeding in tropical birds will greatly contribute to understanding these patterns and their relevance to avian diversity.
Fault bars timing and duration: the power of studying feather fault bars and growth bands together
Authors: Jovani Roger , Diaz‐Real Javier
Published: May 2012

Growth bands and fault bars, widespread features of feathers that form during regeneration, have largely been studied independently. Growth bands result from normal regeneration: each pair of dark/light bands forms every 24 h. Fault bars are a response to stress during regeneration, creating a translucent line that can break the feather. We studied the relative position and width of these two structures in feathers of nestling and adult white storksCiconia ciconia. We first confirmed that one growth band represents 24 h of feather regeneration. Fault bars did not occur at random within growth bands: 65.7% (in nestlings) and 45.6% (in adults) of them occurred in one out of six defined segments within a growth band, namely that segment corresponding to the first one‐third of night time hours. The width of fault bars relative to growth bands suggested that fault bars were produced during a median (range) of 7.0 h (2.7–27.0) in nestlings and 3.7 h (1.8–7.9) in adults. Fault bars were concentrated at feather tips in nestlings, but at central locations in adult feathers. Our results suggest that, in general, fault bars are a discrete event of a finite duration occurring mainly during the night (particularly in nestlings). This, along with current knowledge, suggests that acute stressors, rather than chronic ones, are responsible for fault bar formation. Thus, such acute punctual stressors (a matter of minutes) can have long‐lasting (months−years) physiological effects due to the wing load increase from feather breakage caused by fault bars.