Engineered Human Induced Pluripotent Cells Enable Genetic Code Expansion in Brain Organoids

Human induced pluripotent stem cell (hiPSC) technology has revolutionized studies on human biology. A wide range of cell types and tissue models can be derived from hiPSCs to study complex human diseases. Here, we use PiggyBac-mediated transgenesis to engineer hiPSCs with an expanded genetic code. We demonstrate that genomic integration of expression cassettes for a pyrrolysyl-tRNA synthetase (PylRS), pyrrolysyl-tRNA (PylT) and the target protein of interest enables site-specific incorporation of a non-canonical amino acid (ncAA) in response to an amber stop codon. Neural stem cells, neurons and brain organoids derived from the engineered hiPSCs continue to express the amber suppression machinery and produce ncAA-bearing reporter. The incorporated ncAA can serve as a minimal bioorthogonal handle for further modifications by labeling with fluorescent dyes. Site-directed ncAA mutagenesis will open a wide range of applications to probe and manipulate proteins in brain organoids and other hiPSC-derived cell types and complex tissue models.     

Effect of structure on the glass transition temperatures of linear and crosslinked poly(isobornylacrylate-co-isobutylacrylate)

A series of linear acrylic copolymers based on Isobornyl acrylate (IBOA) and isobutyl acrylate (IsoBA) were elaborated by radical photopolymerization. In addition, several photochemically crosslinked poly(IBOA-co-IsoBA) were prepared by introducing small amounts of 1,6-hexanedioldiacrylate as crosslinking agent. The evolution of the glass transition temperature was determined experimentally by differential scanning calorimetry as a function of composition for both linear and crosslinked poly(IBOA-co-IsoBA), yielding T_g values ranging from (~249) to (~315 K). Theoretical modeling was performed applying Fox, Gordon-Taylor and Couchman-Karasz models by simple calculations using experimental data, leading to only fair agreement between theoretical and experimental values, or by applying fitting procedures involving one or two adjustable parameters. Likewise, the Kwei model, known to take into account hydrogen bonding interactions between monomers, could not describe well the evolution of T_g, indicating the existence of other factors influencing T_g. Finally, a thermodynamic approach based on entropy considerations allowed to attribute an explanation of the evolution of T_g.     

Hohlfaser-Membrankontaktoren zur Stickstoffrückgewinnung aus Prozesswasser einer kommunalen Kläranlage

Process water deriving from digested sludge dewatering at municipal wastewater treatment plants (WWTP) represents an additional load for the plant in terms of nitrogen. Münster WWTP is the first German plant to operate an innovative full-scale membrane contactor facility for nitrogen removal and recovery from process water. Within the scope of this study, aspects influencing the membrane performance and operation as well as utilization of the produced fertilizer are investigated. First results reveal nitrogen removal efficiencies around 85 % and a regional usability of the produced fertilizer.     

A multiplicity of roles for genetic nursing: building toward holistic practice

Nursing's unique contribution to human genetics is holism. There are many roles for basic and advanced practice nurses involved in genetics. Describing these roles makes it easy for nurses to learn more about genetic nursing and helps promote collaboration among professionals. Education for nurses about genetics helps foster greater nursing participation in human genetics. The goal is for nurses to be informed about genetics so that they can better understand people as whole persons whose growth and development, human response patterns, and biopsychosocial processes are affected by and have an effect on human DNA.     

Female and male personality styles: A typological and developmental analysis.

The personality styles of 137 women and 138 men aged 27 yrs were examined in an ongoing Finnish longitudinal study in which the participants were first assessed at age 8. Data were collected by means of a mailed questionnaire, personality inventories, and a semistructured interview. Variables covered personality characteristics, life orientation, and behavioral activities. Both women and men fell into two major clusters, the adjusted (3/4) and the conflicted (1/4). Both clusters divided into subclusters; altogether, 7 were extracted for women and men replicating the personality types obtained by J. Block (1971) despite the use of a different methodology in a different culture. The clusters had roots in individuals' emotional and behavioral regulation from the early school years onward, and they also predicted personality characteristics over a period of 6 years when the Big Five personality factors were used as criteria. (PsycINFO Database Record (c) 2011 APA, all rights reserved)     

Identification of a steroidogenic neurohormone in female mosquitoes

In the female mosquito, Aedes aegypti, neurohormones are released from the brain in response to a blood meal and stimulate the ovaries to secrete ecdysteroid hormones, which modulate yolk protein synthesis in the fat body. Neuropeptides with this bioactivity were isolated from head extracts, and partial sequences from these peptides when aligned gave a 31-residue sequence at the amino terminus. Oligonucleotide primers for this sequence were used to amplify with the polymerase chain reaction a genomic DNA product that hybridized to a clone from a head cDNA library. The cDNA encodes a 149-residue preprohormone that is processed into an 86-residue peptide, as indicated by the mass value obtained from the native peptide, with the expected amino-terminal sequence. After modification, the cDNA for the putative neurohormone was expressed in a bacterial system, and the purified peptide had high specific activity in bioassays, thus confirming that it is a steroidogenic gonadotropin, the first to be identified for invertebrates.     

Personal control over development, identity formation, and future orientation as components of life orientation: A developmental approach.

Two components of life orientation (LO) emerged in a study of 145 men and 142 women at age 26: (1) Positive Orientation (PO), which included self-percepts of personal control over development, positive evaluation of the future, and identity achievement; and (2) Motivated Questioning (MQ), which consisted of identity exploration, motivated future orientation, and dissatisfaction with present achievements. Methods used were an identity status interview, a personal control inventory, and a semistructured interview. In comparison with MQ, PO was more strongly associated with variables involved in adaptive psychological functioning: personal well-being, broad range of goals, social acceptance, and socially constructive ends (A. S. Waterman, 1992). School success studied at ages 8 and 14 and SES contributed to both PO and MQ more strongly in women than in men. LO in men was more dependent on current variables involved in adaptive psychological functioning. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

14C]2-deoxyglucose uptake in the brain of the ring dove (Streptopelia risoria). II. Differential uptake at the onset of incubation

Brain areas involved in the expression of incubation behaviour were identified in male ring dove at the onset of incubation using quantitative [14C]2-deoxyglucose (2DG) autoradiography. The uptake of 2DG in twenty-eight areas found in the hypothalamus and thalamus, and six areas in the forebrain were compared in control non-breeding and incubating birds. Although 2DG utilisation varied significantly between brain areas, significant differences between non-breeding males and incubating males were observed only in 4 of them. These were the nucleus tuberis, nucleus preopticus medialis, nucleus ovoidalis and paleostriatum primitivum. The uptake of 2DG was increased at the onset of incubation in the nucleus tuberis, nucleus preopticus medialis and nucleus ovoidalis and decrease in the paleostriatum primitivum. These findings are consistent with the hypothesis that these brain areas play a role in the initiation of incubation behaviour.     

Measuring validity in sensory analysis

Two methods for assessor evaluation are demonstrated, both based on graphical techniques: plots of an assessor's ability to detect differences vs. his/her repeatability, and the ‘egg-shell plot’ which highlights an assessor's agreement with the panel's rankings. The methods presented in this paper are modifications of already established methods in the area. All methods are illustrated by an example from sensory profiling of beef boullion.     

WASP: A WSI Associative String Processor

ASP (Associative String Processor) modules (and support software) comprise highly-versatile and fault-tolerant building-blocks for the simple construction of dynamically-reconfigurable low-MIMD/high-SIMD second-generation Massively Parallel Processor (MPP) systems. Indeed, based on state-of-the-art microelectronics and packaging technologies, ASP modules constitute a family of packaged MPP configurations for the cost-effective implementation of highly-compact application-specific high performance information processing systems.Based on scalar-vector content-matching rather than location addressing, ASP substrings comprise homogeneous fine-grain SIMD MPP structures, which, in operation, execute a form of set processing (i.e. a sequence of scalar-vector and vector-vector processes) on relevant data. Moreover, application flexibility enables simple tailoring of parallel processing power to match user requirements.WASP devices are WSI (Wafer Scale Integration) implementations of ASP substrings and, as such, constitute fundamental building blocks for the assembly of ASP modules. Exploiting either monolithic or hybrid 1 m CMOS WSI technologies, 8,192-processor WASP devices would enable the assembly of 65,536-processor SEM-E compatible ASP modules achieving 10 Tera-OPS/ft³, 1 Giga-OPS/W and 1 Mega-OPS/$ in cost-effectiveness.The paper discusses second-generation MPP design targets and describes ASP modules for real-time signal and data processing applications. In particular, the paper focuses on the architecture, operation, and implementation of the WASP device and reports on the progress of its development.