DEVELOPMENT OF VIRTUAL EVENT MARKETING

In the last few years the Pichia pastoris expression system has been gaining more and more interest for the expression of recombinant proteins. Many groups have employed fermentation technology in their investigations because the system is fairly easy to scale up and suitable for the production in the milligram to gram range. A large number of heterologous proteins from different sources has been expressed, but the fermentation process technology has been investigated to a lesser extent. A large number of fermentations are carried out in standard bioreactors that may be insufficiently equipped to meet the demands of high-cell-density fermentations of methylotrophic yeasts. In particular, the lack of on-line methanol analysis leads to fermentation protocols that may impair the optimal expression of the desired products. We have used a commercially available methanol sensor to investigate in detail the effects of supplementary glycerol feeding while maintaining a constant methanol concentration during the induction of a Mut+ strain of Pichia pastoris. Specific glycerol feed rates in the range of 38-4.2 mg × g(exp -1) × h(exp -1) (mg glycerol per gram fresh weight per hour) were investigated. Expression of the recombinant scFv antibody fragment was only observed at specific feed rates below 6 mg × g(exp -1) × h(exp -1). At low specific feed rates, growth was even lower than with methanol as the sole carbon source and the harvest expression level of the scFv was only half of that found in the control fermentation. These results show that glycerol inhibits expression driven by the AOX1 promoter even at extremely limited availability and demonstrate the benefits of on-line methanol control in Pichia fermentation research

Quantitative Models and Implicit Complexity

We give new proofs of soundness (all representable functions on base types lies in certain complexity classes) for Elementary Affine Logic, LFPL (a language for polytime computation close to realistic functional programming introduced by one of us), Light Affine Logic and Soft Affine Logic. The proofs are based on a common semantical framework which is merely instantiated in four different ways. The framework consists of an innovative modification of realizability which allows us to use resource-bounded computations as realisers as opposed to including all Turing computable functions as is usually the case in realizability constructions. For example, all realisers in the model for LFPL are polynomially bounded computations whence soundness holds by construction of the model. The work then lies in being able to interpret all the required constructs in the model. While being the first entirely semantical proof of polytime soundness for light logi cs, our proof also provides a notable simplification of the original already semantical proof of polytime soundness for LFPL. A new result made possible by the semantic framework is the addition of polymorphism and a modality to LFPL thus allowing for an internal definition of inductive datatypes.Comment: 29 page

Hydrated Electron Dynamics at a Five Femtosecond Time Scale

Hydrated electrons are studied by frequency resolved pump probe with 5 fs time resolution in the spectral range from 600 nm to 1000 nm. A recurrence detected in the pumpprobe signal at —40 fs is tentatively assigned to coupling to librational motions in the electron's solvent cage

Accuracy Assessments of Cloud Droplet Size Retrievals from Polarized Reflectance Measurements by the Research Scanning Polarimeter

We present an algorithm for the retrieval of cloud droplet size distribution parameters (effective radius and variance) from the Research Scanning Polarimeter (RSP) measurements. The RSP is an airborne prototype for the Aerosol Polarimetery Sensor (APS), which was on-board of the NASA Glory satellite. This instrument measures both polarized and total reflectance in 9 spectral channels with central wavelengths ranging from 410 to 2260 nm. The cloud droplet size retrievals use the polarized reflectance in the scattering angle range between 135deg and 165deg, where they exhibit the sharply defined structure known as the rain- or cloud-bow. The shape of the rainbow is determined mainly by the single scattering properties of cloud particles. This significantly simplifies both forward modeling and inversions, while also substantially reducing uncertainties caused by the aerosol loading and possible presence of undetected clouds nearby. In this study we present the accuracy evaluation of our algorithm based on the results of sensitivity tests performed using realistic simulated cloud radiation fields

MINSTED nanoscopy enters the Ångström localization range

We report all-optical, room-temperature localization of fluorophores with precision in the Ångström range. These precisions are attained in a STED microscope, by encircling the fluorophore with the low-intensity edge of the STED donut beam, while constantly increasing the absolute donut power. Individual fluorophores bound to a DNA strand are localized with σ = 4.7 Å, corresponding to a fraction of the fluorophore size, with only 2,000 detected photons. MINSTED fluorescence nanoscopy with single-digit nanometer resolution is exemplified by imaging nuclear pore complexes and the distribution of nuclear lamin in mammalian cells labeled by transient DNA hybridization. Since our experiments yield a localization precision σ = 2.3 Å, estimated for 10,000 detected photons, we anticipate that MINSTED will open up entirely new areas of application in the study of macromolecular complexes in cells

AI-based structure-function correlation in age-related macular degeneration

Sensitive and robust outcome measures of retinal function are pivotal for clinical trials in age-related macular degeneration (AMD). A recent development is the implementation of artificial intelligence (AI) to infer results of psychophysical examinations based on findings derived from multimodal imaging. We conducted a review of the current literature referenced in PubMed and Web of Science among others with the keywords 'artificial intelligence' and 'machine learning' in combination with 'perimetry', 'best-corrected visual acuity (BCVA)', 'retinal function' and 'age-related macular degeneration'. So far AI-based structure-function correlations have been applied to infer conventional visual field, fundus-controlled perimetry, and electroretinography data, as well as BCVA, and patient-reported outcome measures (PROM). In neovascular AMD, inference of BCVA (hereafter termed inferred BCVA) can estimate BCVA results with a root mean squared error of ~7-11 letters, which is comparable to the accuracy of actual visual acuity assessment. Further, AI-based structure-function correlation can successfully infer fundus-controlled perimetry (FCP) results both for mesopic as well as dark-adapted (DA) cyan and red testing (hereafter termed inferred sensitivity). Accuracy of inferred sensitivity can be augmented by adding short FCP examinations and reach mean absolute errors (MAE) of ~3-5 dB for mesopic, DA cyan and DA red testing. Inferred BCVA, and inferred retinal sensitivity, based on multimodal imaging, may be considered as a quasi-functional surrogate endpoint for future interventional clinical trials in the future

Detecting genomic indel variants with exact breakpoints in single- and paired-end sequencing data using SplazerS

Motivation: The reliable detection of genomic variation in resequencing data is still a major challenge, especially for variants larger than a few base pairs. Sequencing reads crossing boundaries of structural variation carry the potential for their identification, but are difficult to map. Results: Here we present a method for ‘split’ read mapping, where prefix and suffix match of a read may be interrupted by a longer gap in the read-to-reference alignment. We use this method to accurately detect medium-sized insertions and long deletions with precise breakpoints in genomic resequencing data. Compared with alternative split mapping methods, SplazerS significantly improves sensitivity for detecting large indel events, especially in variant-rich regions. Our method is robust in the presence of sequencing errors as well as alignment errors due to genomic mutations/divergence, and can be used on reads of variable lengths. Our analysis shows that SplazerS is a versatile tool applicable to unanchored or single-end as well as anchored paired-end reads. In addition, application of SplazerS to targeted resequencing data led to the interesting discovery of a complete, possibly functional gene retrocopy variant. Availability: SplazerS is available from http://www.seqan.de/projects/ splazers

Psychoanalysis as the Patient: High in Feeling, Low in Energy

This paper examines the increasingly important role that affect is assuming in psychoanalytic research and practice. This rise in the centrality of affect has been at the expense of an independent role for motivation and a dismissal of any energy concept. Difficulties with this affect-first approach are identified and an alternative offered that accords motivation an independent role and accommodates a useful energy concept. Research on esophageal atresia, addiction, and infant suckling are cited in support of this position.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66624/2/10.1177_00030651970450031101.pd