bluuebunny/crossref_metadata_2025_split huggingface dataset:
A novel fed-batch based cultivation method provides high cell-density and improves yield of soluble recombinant proteins in shaken cultures
Authors: Krause Mirja , Ukkonen Kaisa , Haataja Tatu , Ruottinen Maria , Glumoff Tuomo , Neubauer Antje , Neubauer Peter , Vasala Antti 
Published: February 2010

Abstract

Background
Cultivations for recombinant protein production in shake flasks should provide high cell densities, high protein productivity per cell and good protein quality. The methods described in laboratory handbooks often fail to reach these goals due to oxygen depletion, lack of pH control and the necessity to use low induction cell densities. In this article we describe the impact of a novel enzymatically controlled fed-batch cultivation technology on recombinant protein production in Escherichia coli in simple shaken cultures.

Results
The enzymatic glucose release system together with a well-balanced combination of mineral salts and complex medium additives provided high cell densities, high protein yields and a considerably improved proportion of soluble proteins in harvested cells. The cultivation method consists of three steps: 1) controlled growth by glucose-limited fed-batch to OD600 ~10, 2) addition of growth boosters together with an inducer providing efficient protein synthesis within a 3 to 6 hours period, and 3) a slow growth period (16 to 21 hours) during which the recombinant protein is slowly synthesized and folded. Cell densities corresponding to 10 to 15 g l-1 cell dry weight could be achieved with the developed technique. In comparison to standard cultures in LB, Terrific Broth and mineral salt medium, we typically achieved over 10-fold higher volumetric yields of soluble recombinant proteins.

Conclusions
We have demonstrated that by applying the novel EnBase® Flo cultivation system in shaken cultures high cell densities can be obtained without impairing the productivity per cell. Especially the yield of soluble (correctly folded) proteins was significantly improved in comparison to commonly used LB, Terrific Broth or mineral salt media. This improvement is thought to result from a well controlled physiological state during the whole process. The higher volumetric yields enable the use of lower culture volumes and can thus significantly reduce the amount of time and effort needed for downstream processing or process optimization. We claim that the new cultivation system is widely applicable and, as it is very simple to apply, could widely replace standard shake flask approaches.
Analysis of an alternative human CD133 promoter reveals the implication of Ras/ERK pathway in tumor stem-like hallmarks
Authors: Tabu Kouichi , Kimura Taichi , Sasai Ken , Wang Lei , Bizen Norihisa , Nishihara Hiroshi , Taga Tetsuya , Tanaka Shinya 
Published: February 2010

Abstract

Background
An increasing number of studies support the presence of stem-like cells in human malignancies. These cells are primarily responsible for tumor initiation and thus considered as a potential target to eradicate tumors. CD133 has been identified as an important cell surface marker to enrich the stem-like population in various human tumors. To reveal the molecular machinery underlying the stem-like features in tumor cells, we analyzed a promoter of CD133 gene using human colon carcinoma Caco-2 and synovial sarcoma Fuji cells, which endogenously express CD133 gene.

Results
A reporter analysis revealed that P5 promoter, located far upstream in a human CD133 gene locus, exhibits the highest activity among the five putative promoters (P1 to P5). Deletion and mutation analysis identified two ETS binding sites in the P5 region as being essential for its promoter activity. Electrophoretic mobility shift assays demonstrated the specific binding between nuclear factors and the ETS binding sequence. Overexpression of dominant-negative forms of Ets2 and Elk1 resulted in the significant decrease of P5 activity. Furthermore, treatment of Fuji cells with a specific MEK/ERK inhibitor, U0126, also markedly decreased CD133 expression, but there was no significant effect in Caco-2 cells, suggesting cell type-specific regulation of CD133 expression. Instead, the side population, another hallmark of TSLCs, was dramatically diminished in Caco-2 cells by U0126. Finally, Ras-mediated oncogenic transformation in normal human astrocytes conferred the stem-like capability to form neurosphere-like colonies with the increase of CD133 mRNA expression.

Conclusions
In conclusion, the Ras/ERK pathway at least in part contributes to the maintenance and the acquisition of stem-like hallmarks, although the extent of its contribution is varied in a cell type-specific manner. These findings could help our comprehensive understanding of tumor stemness, and also improve the development of eradicative therapies against human malignancies.
Anti-inflammatory activity of nanocrystalline silver-derived solutions in porcine contact dermatitis
Authors: Nadworny Patricia L , Wang JianFei , Tredget Edward E , Burrell Robert E 
Published: February 2010

AbstractBackgroundNanocrystalline silver dressings have anti-inflammatory activity, unlike solutions containing Ag+only, which may be due to dissolution of multiple silver species. These dressings can only be used to treat surfaces. Thus, silver-containing solutions with nanocrystalline silver properties could be valuable for treating hard-to-dress surfaces and inflammatory conditions of the lungs and bowels. This study tested nanocrystalline silver-derived solutions for anti-inflammatory activity.MethodsInflammation was induced on porcine backs using dinitrochlorobenzene. Negative and positive controls were treated with distilled water. Experimental groups were treated with solutions generated by dissolving nanocrystalline silver in distilled water adjusted to starting pHs of 4 (using CO2), 5.6 (as is), 7, and 9 (using Ca(OH)2). Solution samples were analyzed for total silver. Daily imaging, biopsying, erythema and oedema scoring, and treatments were performed for three days. Biopsies were processed for histology, immunohistochemistry (for IL-4, IL-8, IL-10, TNF-α, EGF, KGF, KGF-2, and apoptotic cells), and zymography (MMP-2 and -9). One-way ANOVAs with Tukey-Kramer post tests were used for statistical analyses.ResultsAnimals treated with pH 7 and 9 solutions showed clear visual improvements. pH 9 solutions resulted in the most significant reductions in erythema and oedema scores. pH 4 and 7 solutions also reduced oedema scores. Histologically, all treatment groups demonstrated enhanced re-epithelialisation, with decreased inflammation. At 24 h, pMMP-2 expression was significantly lowered with pH 5.6 and 9 treatments, as was aMMP-2 expression with pH 9 treatments. In general, treatment with silver-containing solutions resulted in decreased TNF-α and IL-8 expression, with increased IL-4, EGF, KGF, and KGF-2 expression. At 24 h, apoptotic cells were detected mostly in the dermis with pH 4 and 9 treatments, nowhere with pH 5.6, and in both the epidermis and dermis with pH 7. Solution anti-inflammatory activity did not correlate with total silver content, as pH 4 solutions contained significantly more silver than all others.ConclusionsNanocrystalline silver-derived solutions appear to have anti-inflammatory/pro-healing activity, particularly with a starting pH of 9. Solutions generated differently may have varying concentrations of different silver species, only some of which are anti-inflammatory. Nanocrystalline silver-derived solutions show promise for a variety of anti-inflammatory treatment applications.
Oxygen consumption is depressed in patients with lactic acidosis due to biguanide intoxication
Authors: Protti Alessandro , Russo Riccarda , Tagliabue Paola , Vecchio Sarah , Singer Mervyn , Rudiger Alain , Foti Giuseppe , Rossi Anna , Mistraletti Giovanni , Gattinoni Luciano 
Published: February 2010

Abstract
Introduction
Lactic acidosis can develop during biguanide (metformin and phenformin) intoxication, possibly as a consequence of mitochondrial dysfunction. To verify this hypothesis, we investigated whether body oxygen consumption (VO2), that primarily depends on mitochondrial respiration, is depressed in patients with biguanide intoxication.

Methods
Multicentre retrospective analysis of data collected from 24 patients with lactic acidosis (pH 6.93 ± 0.20; lactate 18 ± 6 mM at hospital admission) due to metformin (n = 23) or phenformin (n = 1) intoxication. In 11 patients, VO2 was computed as the product of simultaneously recorded arterio-venous difference in O2 content [C(a-v)O2] and cardiac index (CI). In 13 additional cases, C(a-v)O2, but not CI, was available.

Results
On day 1, VO2 was markedly depressed (67 ± 28 ml/min/m2) despite a normal CI (3.4 ± 1.2 L/min/m2). C(a-v)O2 was abnormally low in both patients either with (2.0 ± 1.0 ml O2/100 ml) or without (2.5 ± 1.1 ml O2/100 ml) CI (and VO2) monitoring. Clearance of the accumulated drug was associated with the resolution of lactic acidosis and a parallel increase in VO2 (P < 0.001) and C(a-v)O2 (P < 0.05). Plasma lactate and VO2 were inversely correlated (R2 0.43; P < 0.001, n = 32).

Conclusions
VO2 is abnormally low in patients with lactic acidosis due to biguanide intoxication. This finding is in line with the hypothesis of inhibited mitochondrial respiration and consequent hyperlactatemia.
Auditory Attraction: Activation of Visual Cortex by Music and Sound in Williams Syndrome
Authors: Thornton-Wells Tricia A. , Cannistraci Christopher J. , Anderson Adam W. , Kim Chai-Youn , Eapen Mariam , Gore John C. , Blake Randolph , Dykens Elisabeth M. 
Published: February 2010

AbstractWilliams syndrome is a genetic neurodevelopmental disorder with a distinctive phenotype, including cognitive–linguistic features, nonsocial anxiety, and a strong attraction to music. We performed functional MRI studies examining brain responses to musical and other types of auditory stimuli in young adults with Williams syndrome and typically developing controls. In Study 1, the Williams syndrome group exhibited unforeseen activations of the visual cortex to musical stimuli, and it was this novel finding that became the focus of two subsequent studies. Using retinotopy, color localizers, and additional sound conditions, we identified specific visual areas in subjects with Williams syndrome that were activated by both musical and nonmusical auditory stimuli. The results, similar to synesthetic-like experiences, have implications for cross-modal sensory processing in typical and atypical neurodevelopment.
Iothalamate Quantification by Tandem Mass Spectrometry to Measure Glomerular Filtration Rate
Authors: Seegmiller Jesse C , Burns Bradley E , Fauq Abdul H , Mukhtar Naveen , Lieske John C , Larson Timothy S 
Published: February 2010

Abstract
Background: Glomerular filtration rate (GFR) can be determined by measuring renal clearance of the radiocontrast agent iothalamate. Current analytic methods for quantifying iothalamate concentrations in plasma and urine using liquid chromatography or capillary electrophoresis have limitations such as long analysis times and susceptibility to interferences. We developed a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method to overcome these limitations.
Methods: Urine and plasma samples were deproteinized using acetonitrile and centrifugation. The supernatant was diluted in water and analyzed by LC-MS/MS using a water:methanol gradient. We monitored 4 multiple reaction monitoring transitions: m/z 614.8–487.0, 614.8–456.0, 614.8–361.1, and 614.8–177.1. We compared the results to those obtained via our standard capillary electrophoresis (CE-UV) on samples from 53 patients undergoing clinical GFR testing.
Results: Mean recovery was 90%–110% in both urine and plasma matrices. Imprecision was ≤15% for the m/z 614.8–487.0 and 614.8–456.0 transitions over a 10-day period at 1 mg/L. Method comparison for 159 patient samples (53 clearances) provided the following Passing–Bablok regressions: plasma iothalamate LC-MS/MS (y) vs CE-UV (x), y = 0.99x + 0.36; urine iothalamate LC-MS/MS vs CE-UV, y = 1.01x + 0.31; corrected GFR LC-MS/MS vs CE-UV, y = 1.00x + 0.00. Interfering substances prevented accurate iothalamate quantification by CE-UV in 2 patients, whereas these samples could be analyzed by LC-MS/MS.
Conclusions: Iothalamate can be quantified by LC-MS/MS for GFR measurement. This method circumvents potential problems with interfering substances that occasionally confound accurate GFR determinations.
Trends in Childhood Leukemia in Basrah, Iraq, 1993–2007
Authors: Hagopian Amy , Lafta Riyadh , Hassan Jenan , Davis Scott , Mirick Dana , Takaro Tim 
Published: February 2010

Objectives. Through a sister-university relationship between the University of Basrah and the University of Washington, we analyzed Ibn Ghazwan Hospital's leukemia registry data to evaluate trends in childhood leukemia since 1993. Methods. We documented leukemia cases among children aged 0 to 14 years for each of the last 15 years. Population data were obtained from a 1997 census and various subsequent estimates to calculate rates. Results. We observed 698 cases of childhood leukemia between 1993 and 2007, ranging between 15 cases (2.6 per 100 000 annual rate) in the first year and 56 cases (6.9 per 100 000 annual rate) in the final year, reaching a peak of 97 cases in 2006 (12.2 per 100 000 annual rate). Conclusions. Childhood leukemia rates in Basrah more than doubled over a 15-year period. The test for trend was significant (P = .03). Basrah's childhood leukemia rate compared unfavorably with neighboring Kuwait and nearby Oman, as well as the United States, the European Union, and other countries.
Effect of a Two-Year Obesity Prevention Intervention on Percentile Changes in Body Mass Index and Academic Performance in Low-Income Elementary School Children
Authors: Hollar Danielle , Messiah Sarah E. , Lopez-Mitnik Gabriela , Hollar T. Lucas , Almon Marie , Agatston Arthur S. 
Published: February 2010

Objectives. We assessed the effects of a school-based obesity prevention intervention that included dietary, curricula, and physical activity components on body mass index (BMI) percentiles and academic performance among low-income elementary school children. Methods. The study had a quasi-experimental design (4 intervention schools and 1 control school; 4588 schoolchildren; 48% Hispanic) and was conducted over a 2-year period. Data are presented for the subset of the cohort who qualified for free or reduced-price school lunches (68% Hispanic; n = 1197). Demographic and anthropometric data were collected in the fall and spring of each year, and academic data were collected at the end of each year. Results. Significantly more intervention than control children stayed within normal BMI percentile ranges both years (P = .02). Although not significantly so, more obese children in the intervention (4.4%) than in the control (2.5%) decreased their BMI percentiles. Overall, intervention schoolchildren had significantly higher math scores both years (P < .001). Hispanic and White intervention schoolchildren were significantly more likely to have higher math scores (P < .001). Although not significantly so, intervention schoolchildren had higher reading scores both years. Conclusions. School-based interventions can improve health and academic performance among low-income schoolchildren.
Three-Phase Measurements of Non-wetting Phase Trapping in Unconsolidated Sand Packs
Authors: Al Mansoori Saleh K. , Pentland Christopher H. , Iglauer Stefan , Blunt Martin J. 
Published: February 2010

Abstract
We perform a series of experiments in water-wet sand packs to measure the trapped saturations of oil and gas as a function of initial saturation. We start with brine-saturated columns and inject octane (oil) to reach irreducible water saturation followed by displacement by air (gas) from the top, allowing oil and air to drain under gravity for different amounts of time, then finally brine is injected from the bottom to trap both oil and gas. The columns are sliced and a sensitive and accurate measurement of saturation along the column is made using gas chromatography. The maximum residual gas saturation is over 20%, compared to 14% for two-phase flow (Al Mansoori et al. 2009). For lower initial gas saturation, the amount of trapping is similar to that reached in an equivalent two-phase experiment. We also find that the amount of oil trapped is insensitive to either the initial gas saturation or the amount of gas that is trapped. More oil is trapped than would be predicted from an equivalent two-phase system, although the trapped saturation is never larger than the maximum reached in two-phase flow (around 11%) (Pentland et al. 2008). These initially surprising results are explained in the context of oil layer stability and the competition between snap-off and piston-like advance. In unconsolidated two-phase water-wet systems, displacement is principally by cooperative piston-like advance with relatively little trapping, whereas in consolidated media snap-off is generally more significant. However, during three-phase waterflooding, oil layer collapse events rapidly trap the oil which acts as a barrier to direct water-gas displacement, except by snap-off, leading to enhanced gas trapping.
Preliminary malacological investigation on the loess profile at Zmajevac, Croatia
Authors: Molnár David , Hupuczi Júlia , Galović Lidija , Sümegi Pál 
Published: February 2010

Abstract
More than 5 500 specimens of 37 terrestrial snail species were collected from a loess paleosol section at Zmajevac village, Croatia. The presence of Ena montana, Mastus bielzi, Cochlodina laminata, Macrogastra ventricosa, Clausilia pumila, Clausilia dubia, Pseudofusulus varians, Trichia unidentata and Trichia edentula species from the lower paleosol layer suggests that this malacological zone formed during the initial phase of a Middle Pleistocene interglacial cycle.
Metabolite concentrations in supraventricular white matter from teenage to early old age: A short echo time 1h magnetic resonance spectroscopy (MRS) study
Authors: Raininko Raili , Mattsson Peter 
Published: February 2010

Background: Age- and sex-related changes of metabolites in healthy adult brains have been examined with different 1H magnetic resonance spectroscopy (MRS) methods in varying populations, and with differing results. A long repetition time and short echo time technique reduces quantification errors due to T1 and T2 relaxation effects and makes it possible to measure metabolites with short T2 relaxation times. Purpose: To examine the effect of age on the metabolite concentrations measured by 1H MRS in normal supraventricular white matter using a long repetition time (TR) and a short echo time (TE). Material and Methods: Supraventricular white matter of 57 healthy subjects (25 women, 32 men), aged 13 to 72 years, was examined with a single-voxel MRS at 1.5T using a TR of 6000 ms and a TE of 22 ms. Tissue water was used as a reference in quantification. Results: Myoinositol increased slightly and total N-acetyl aspartate (NAA) decreased slightly with increasing age. Glutamine/glutamate complex (Glx) showed U-shaped age dependence, with highest concentrations in the youngest and oldest subjects. No significant age dependence was found in total choline and total creatine. No gender differences were found. Macromolecule/ lipid (ML) fractions were reliably measurable only in 36/57 or even fewer subjects and showed very large deviations. Conclusion: The concentrations of several metabolites in cerebral supraventricular white matter are age dependent on 1H MRS, even in young and middle-aged people, and age dependency can be nonlinear. Each 1H MRS study of the brain should therefore take age into account, whereas sex does not appear to be so important. The use of macromolecule and lipid evaluations is compromised by less successful quantification and large variations in healthy people.
Tempest in the Tea Leaves: Broadcasting the Esoteric Arts and Mystic Sciences, 1937–1953
Authors: Kuffert Len 
Published: February 2010

Abstract: From 1937 until 1953, it was contrary to Canadian broadcasting regulations to broadcast programs with speakers claiming to have supernatural abilities, or to offer personal advice over the airwaves. As the broadcast regulator, the Canadian Broadcasting Corporation was responsible for enforcing the ban, which meant preventing privately operated stations from producing such programs. The struggle over what sort of programming was suitable not only mirrored the differences between audiences and regulators or broadcasters and regulators, but also attitudes toward esoteric lifestyles, religion, science, and the place of entertainment on the air. Regulators uncomfortable with the potential for exploitation or exposure of listeners' private affairs repeatedly turned back attempts by broadcasters to circumvent the regulation, and in the process revealed some deep-seated class and gender-based assumptions about radio's early audiences.
‘A Species of Treason?’: Deportation and Nation-Building in the Case of Tomo Čačić, 1931–1934
Authors: Molinaro Dennis G. 
Published: February 2010

Abstract: Deportation was used to remove political radicals from Canada during the early 1930s, as well as immigrants receiving poverty or unemployment relief. Studies of deportation in North America are limited and have rarely focused on ideas of ‘the nation’ as a primary focus of debates over deportation; and they have seldom dwelt upon the local efforts of those resisting deportation. Arguably the most influential works in this area are Barbara Roberts's Whence They Came: Deportation in Canada 1900–1935 and Anthony Rasporich's ‘Tomo Čačić: Rebel without a Country.’ This paper builds on the work of both authors by explaining not only the mechanisms, but also the ideological moorings, of Depression-era deportation policies, through the revealing case study of Tomo Čačić, a transnational radical who thought globally and acted locally. The power to deport provided the state with a powerful nation-shaping tool. Deportation of political radicals served a dual purpose: it both isolated and removed those identified as political enemies of the political order and, conversely, demonstrated by implication the qualities of those who were qualified to figure as citizens within the emerging Canadian nation.
Parasites from ‘Alien Shores’: The Decline of Canada's Freshwater Fishing Industry
Authors: Piper Liza 
Published: February 2010

Abstract: After 1900, the Northwest fishing industry expanded substantially from southern Manitoba to the Northwest Territories, with production directed principally to American urban markets. In response to the economic crisis of the 1930s, new international trade regulations prohibited the importation of fish infected with the Triaenophorus parasite and included two of the most important commercial freshwater fish species: tullibee and lake whitefish. In the following decades, in spite of repeated state and scientific interventions in the fishery, Canada's freshwater fishing industry was unable to fully recover from the economic impacts of the new trade regulations. This failure reflected the unprecedented influence of the parasite and its ecological relationships to other species and the freshwaters in which it was found. This paper goes beyond a conventional narrative of the impacts of economic protectionism to ascertain how the natural context in which resources such as fish are found intervenes in human socio-economic affairs.