Microfluidic chromatography for early stage evaluation of biopharmaceutical binding and separation conditions

Optimization of separation conditions for biopharmaceuticals requires evaluation of a large number of process variables. To miniaturize this evaluation a microfluidic column (1.5 mu L volume and 1cm height) was fabricated and packed with a typical process scale resin. The device was assessed by comparison to a protein separation at conventional laboratory scale. This was based upon measurement of the quality of packing and generation of breakthrough and elution curves. Dynamic binding capacities from the microfluidic column compared well with the laboratory scale. Microfluidic scale gradient elution separations also equated to the laboratory column three orders of magnitude larger in scale

Ariel - Volume 6 Number 3

Editors Mark Dembert J.D. Kanofsky Frank Chervenak John Lammie Curt Cummings Staff Ken Jaffe Bob Sklaroff Halley Faust Jim Burke Nancy Redfern Hans Weltin Photographer Larry Glazerman Overseas Editor Mike Sinason Humorist Jim McCan

Cyclin D activates the Rb tumor suppressor by mono-phosphorylation

The widely accepted model of G1 cell cycle progression proposes that cyclin D:Cdk4/6 inactivates the Rb tumor suppressor during early G1 phase by progressive multi-phosphorylation, termed hypo-phosphorylation, to release E2F transcription factors. However, this model remains unproven biochemically and the biologically active form(s) of Rb remains unknown. In this study, we find that Rb is exclusively mono-phosphorylated in early G1 phase by cyclin D:Cdk4/6. Mono-phosphorylated Rb is composed of 14 independent isoforms that are all targeted by the E1a oncoprotein, but show preferential E2F binding patterns. At the late G1 Restriction Point, cyclin E:Cdk2 inactivates Rb by quantum hyper-phosphorylation. Cells undergoing a DNA damage response activate cyclin D:Cdk4/6 to generate mono-phosphorylated Rb that regulates global transcription, whereas cells undergoing differentiation utilize un-phosphorylated Rb. These observations fundamentally change our understanding of G1 cell cycle progression and show that mono-phosphorylated Rb, generated by cyclin D:Cdk4/6, is the only Rb isoform in early G1 phase. DOI: http://dx.doi.org/10.7554/eLife.02872.00

High-Yielding Corn Response to Applied Phosphorus, Potassium, and Sulfur in Nebraska

Nutrient management recommendations may change as yield levels and efficiency of crop production increase. Recommendations for P, K, and S were evaluated using results from 34 irrigated corn (Zea mays L.) trials conducted in diverse situations across Nebraska. The mean yield was 14.7 Mg ha-1 with adequate fertilizer applied. Th e median harvest index values were 0.52, 0.89, 0.15, and 0.56 for biomass, P, K, and S, respectively. Median grain yields were 372, 49, and 613 kg kg-1 of above-ground plant uptake of P, K, and S, respectively. The estimated critical Bray-1 P level for corn response to 20 kg P ha-1 was 20 mg kg-1 when the previous crop was corn compared with 10 mg kg-1 when corn followed soybean [Glycine max (L.) Merr.]. Soil test K was generally high with only three site-years kg-1. Over all trials, application of 40 kg K ha-1 resulted in a 0.2 Mg ha-1 mean grain yield decrease. Application of 22 kg S ha-1 did not result in significant yield increase in any trial. Soil test results accounted for twice as much variation in nutrient uptake when soil organic matter (SOM) and pH were considered in addition to the soil test nutrient values. The results indicate a need to revise the current recommendation for P, to maintain the current K and S recommendations, and to use SOM and pH in addition to soil test nutrient values in estimating applied nutrient requirements for irrigated high yield corn production

High-Yielding Corn Response to Applied Phosphorus, Potassium, and Sulfur in Nebraska

Nutrient management recommendations may change as yield levels and efficiency of crop production increase. Recommendations for P, K, and S were evaluated using results from 34 irrigated corn (Zea mays L.) trials conducted in diverse situations across Nebraska. The mean yield was 14.7 Mg ha-1 with adequate fertilizer applied. Th e median harvest index values were 0.52, 0.89, 0.15, and 0.56 for biomass, P, K, and S, respectively. Median grain yields were 372, 49, and 613 kg kg-1 of above-ground plant uptake of P, K, and S, respectively. The estimated critical Bray-1 P level for corn response to 20 kg P ha-1 was 20 mg kg-1 when the previous crop was corn compared with 10 mg kg-1 when corn followed soybean [Glycine max (L.) Merr.]. Soil test K was generally high with only three site-years kg-1. Over all trials, application of 40 kg K ha-1 resulted in a 0.2 Mg ha-1 mean grain yield decrease. Application of 22 kg S ha-1 did not result in significant yield increase in any trial. Soil test results accounted for twice as much variation in nutrient uptake when soil organic matter (SOM) and pH were considered in addition to the soil test nutrient values. The results indicate a need to revise the current recommendation for P, to maintain the current K and S recommendations, and to use SOM and pH in addition to soil test nutrient values in estimating applied nutrient requirements for irrigated high yield corn production

Nonlinear theory of mirror instability near threshold

An asymptotic model based on a reductive perturbative expansion of the drift kinetic and the Maxwell equations is used to demonstrate that, near the instability threshold, the nonlinear dynamics of mirror modes in a magnetized plasma with anisotropic ion temperatures involves a subcritical bifurcation,leading to the formation of small-scale structures with amplitudes comparable with the ambient magnetic field

A nonlinear theory of the parallel firehose and gyrothermal instabilities in a weakly collisional plasma

Weakly collisional plasmas dynamically develop pressure anisotropies with respect to the magnetic field. These anisotropies trigger plasma instabilities at scales just above the ion Larmor radius \rho_i and much below the mean free path \lambda_{mfp}. They have growth rates of a fraction of the ion cyclotron frequency - much faster than either the global dynamics or local turbulence. The instabilities dramatically modify the transport properties and, therefore, the macroscopic dynamics of the plasma. Their nonlinear evolution drives pressure anisotropies towards marginal stability, controlled by the plasma beta \beta_i. Here this nonlinear evolution is worked out for the simplest analytically tractable example - the parallel firehose instability. In the nonlinear regime, both analytical theory and the numerical solution predict secular growth of magnetic fluctuations. They develop a k^{-3} spectrum, extending from scales somewhat larger than \rho_i to the maximum scale that grows secularly with time (~t^{1/2}); the relative pressure anisotropy (\pperp-\ppar)/\ppar tends to the marginal value -2/\beta_i. The marginal state is achieved via changes in the magnetic field, not particle scattering. When a parallel ion heat flux is present, the firehose mutates into the new gyrothermal instability (GTI), which continues to exist up to firehose-stable values of pressure anisotropy, which can be positive and are limited by the heat flux. The nonlinear evolution of the GTI also features secular growth of magnetic fluctuations, but the spectrum is eventually dominated by modes around the scale ~\rho_i l_T/\lambda_{mfp}, where l_T is the scale of the parallel temperature variation. Implications for momentum and heat transport are speculated about. This study is motivated by the dynamics of galaxy cluster plasmas.Comment: 34 pages, replaced with the version published in MNRA

A Randomized Study Comparing Digital Imaging to Traditional Glass Slide Microscopy for Breast Biopsy and Cancer Diagnosis.

BACKGROUND: Digital whole slide imaging may be useful for obtaining second opinions and is used in many countries. However, the U.S. Food and Drug Administration requires verification studies. METHODS: Pathologists were randomized to interpret one of four sets of breast biopsy cases during two phases, separated by ≥9 months, using glass slides or digital format (sixty cases per set, one slide per case, RESULTS: Sixty-five percent of responding pathologists were eligible, and 252 consented to randomization; 208 completed Phase I (115 glass, 93 digital); and 172 completed Phase II (86 glass, 86 digital). Accuracy was slightly higher using glass compared to digital format and varied by category: invasive carcinoma, 96% versus 93% ( CONCLUSIONS: In this large randomized study, digital format interpretations were similar to glass slide interpretations of benign and invasive cancer cases. However, cases in the middle of the spectrum, where more inherent variability exists, may be more problematic in digital format. Future studies evaluating the effect these findings exert on clinical practice and patient outcomes are required

Angiographic Findings and Outcome in Diabetec Patients Treated With Thrombolytic Therapy for Acute Myocardial Infarction: The GUSTO-I Experience

OBJECTIVES: This study sought to determine whether diabetes mellitus, in the setting of thrombolysis for acute myocardial infarction, affects 1) early infarct-related artery patency and reocclusion rates; and 2) global and regional ventricular function indexes. We also sought to assess whether angiographic or baseline clinical variables, or both, can account for the known excess mortality after myocardial infarction in the diabetic population. BACKGROUND: Mortality after acute myocardial infarction in patients with diabetes is approximately twice that of nondiabetic patients. It is uncertain whether this difference in mortality is due to a lower rate of successful thrombolysis, increased reocclusion after successful thrombolysis, greater ventricular injury or a more adverse angiographic or clinical profile in diabetic patients. METHODS: Patency rates and global and regional left ventricular function were determined in patients enrolled in the GUSTO-I Angiographic Trial. Thirty-day mortality differences between those with and without diabetes were compared. RESULTS: The diabetic cohort had a significantly higher proportion of female and elderly patients, and th

Mapping the Hsp90 Genetic Interaction Network in Candida albicans Reveals Environmental Contingency and Rewired Circuitry

The molecular chaperone Hsp90 regulates the folding of diverse signal transducers in all eukaryotes, profoundly affecting cellular circuitry. In fungi, Hsp90 influences development, drug resistance, and evolution. Hsp90 interacts with ∼10% of the proteome in the model yeast Saccharomyces cerevisiae, while only two interactions have been identified in Candida albicans, the leading fungal pathogen of humans. Utilizing a chemical genomic approach, we mapped the C. albicans Hsp90 interaction network under diverse stress conditions. The chaperone network is environmentally contingent, and most of the 226 genetic interactors are important for growth only under specific conditions, suggesting that they operate downstream of Hsp90, as with the MAPK Hog1. Few interactors are important for growth in many environments, and these are poised to operate upstream of Hsp90, as with the protein kinase CK2 and the transcription factor Ahr1. We establish environmental contingency in the first chaperone network of a fungal pathogen, novel effectors upstream and downstream of Hsp90, and network rewiring over evolutionary time