Modularity of PRC1 composition and chromatin interaction define condensate properties.

Polycomb repressive complexes (PRCs) play a key role in gene repression and are indispensable for proper development. Canonical PRC1 forms condensates in vitro and in cells that are proposed to contribute to the maintenance of repression. However, how chromatin and the various subunits of PRC1 contribute to condensation is largely unexplored.

Modularity of PRC1 Composition and Chromatin Interaction define Condensate Properties.

Polycomb repressive complexes (PRC) play a key role in gene repression and are indispensable for proper development. Canonical PRC1 forms condensates and in cells and the ability of PRC1 to form condensates has been proposed to contribute to maintenance of repression. However, how chromatin and the various subunits of PRC1 contribute to condensation is largely unexplored.

Modeling Non-additive Effects in Neighboring Chemically Identical Fluorophores.

Quantitative fluorescence analysis is often used to derive chemical properties, including stoichiometries, of biomolecular complexes. One fundamental underlying assumption in the analysis of fluorescence data─whether it be the determination of protein complex stoichiometry by super-resolution, or step-counting by photobleaching, or the determination of RNA counts in diffraction-limited spots in RNA fluorescence hybridization (RNA-FISH) experiments─is that fluorophores behave identically and do not interact. However, recent experiments on fluorophore-labeled DNA origami structures such as fluorocubes have shed light on the nature of the interactions between identical fluorophores as these are brought closer together, thereby raising questions on the validity of the modeling assumption that fluorophores do not interact.

Three-color single-molecule imaging reveals conformational dynamics of dynein undergoing motility.

The motor protein dynein undergoes coordinated conformational changes of its domains during motility along microtubules. Previous single-molecule studies analyzed the motion of the AAA rings of the dynein homodimer, but not the distal microtubule-binding domains (MTBDs) that step along the track. Here, we simultaneously tracked with nanometer precision two MTBDs and one AAA ring of a single dynein as it underwent hundreds of steps using three-color imaging.

A 6-nm ultra-photostable DNA FluoroCube for fluorescence imaging.

Photobleaching limits extended imaging of fluorescent biological samples. We developed DNA-based 'FluoroCubes' that are similar in size to the green fluorescent protein, have single-point attachment to proteins, have a ~54-fold higher photobleaching lifetime and emit ~43-fold more photons than single organic dyes. We demonstrate that DNA FluoroCubes provide outstanding tools for single-molecule imaging, allowing the tracking of single motor proteins for >800 steps with nanometer precision.

Coupling of ATPase activity, microtubule binding, and mechanics in the dynein motor domain.

The movement of a molecular motor protein along a cytoskeletal track requires communication between enzymatic, polymer-binding, and mechanical elements. Such communication is particularly complex and not well understood in the dynein motor, an ATPase that is comprised of a ring of six AAA domains, a large mechanical element (linker) spanning over the ring, and a microtubule-binding domain (MTBD) that is separated from the AAA ring by a ~ 135 Å coiled-coil stalk. We identified mutations in the stalk that disrupt directional motion, have microtubule-independent hyperactive ATPase activity, and nucleotide-independent low affinity for microtubules.

Nanometer-accuracy distance measurements between fluorophores at the single-molecule level.

Light microscopy is a powerful tool for probing the conformations of molecular machines at the single-molecule level. Single-molecule Förster resonance energy transfer can measure intramolecular distance changes of single molecules in the range of 2 to 8 nm. However, current superresolution measurements become error-prone below 25 nm.

Determination of ubiquitin fitness landscapes under different chemical stresses in a classroom setting.

Ubiquitin is essential for eukaryotic life and varies in only 3 amino acid positions between yeast and humans. However, recent deep sequencing studies indicate that ubiquitin is highly tolerant to single mutations. We hypothesized that this tolerance would be reduced by chemically induced physiologic perturbations.

Folding complex DNA nanostructures from limited sets of reusable sequences.

Scalable production of DNA nanostructures remains a substantial obstacle to realizing new applications of DNA nanotechnology. Typical DNA nanostructures comprise hundreds of DNA oligonucleotide strands, where each unique strand requires a separate synthesis step. New design methods that reduce the strand count for a given shape while maintaining overall size and complexity would be highly beneficial for efficiently producing DNA nanostructures.

Pain management in the neonatal piglet during routine management procedures. Part 2: grading the quality of evidence and the strength of recommendations.

Piglets reared in swine production in the USA undergo painful procedures that include castration, tail docking, teeth clipping, and identification with ear notching or tagging. These procedures are usually performed without pain mitigation. The objective of this project was to develop recommendations for pain mitigation in 1- to 28-day-old piglets undergoing these procedures.

Pain management in the neonatal piglet during routine management procedures. Part 1: a systematic review of randomized and non-randomized intervention studies.

Routine procedures carried out on piglets (i.e. castration, tail docking, teeth clipping, and ear notching) are considered painful.

Increasing the statistical significance of entanglement detection in experiments.

Entanglement is often verified by a violation of an inequality like a Bell inequality or an entanglement witness. Considerable effort has been devoted to the optimization of such inequalities in order to obtain a high violation. We demonstrate theoretically and experimentally that such an optimization does not necessarily lead to a better entanglement test, if the statistical error is taken into account.

Space allowance for dry, pregnant sows in pens: body condition, skin lesions, and performance.

Different floor space allowances for dry, pregnant sows in pens were evaluated to determine the impacts of space on sow performance, productivity, and body lesions during 2 consecutive farrowings. Treatment groups of 5 sows/pen were assigned to 1.4, 2.

Immune responses of piglets to weaning stress: impacts of photoperiod.

An acute stress response can be provoked by abrupt social, nutritional, and environmental changes associated with weaning, and this may disrupt homeostasis and thus compromise well-being. Manipulating environmental factors, such as photoperiod, might provide a simple way to reduce the physiological consequences that piglets experience due to weaning stress. The objective of this study was to evaluate the impacts of photoperiod manipulation across various weaning ages on leukocyte populations, lymphocyte proliferation, natural killer cytotoxicity (NK), chemotaxis, phagocytosis, and immunoglobulin G, cortisol, and BW of piglets during the nursery phase.

Heat and social rank impact behavior and physiology of PRRS-virus-infected pigs.

Changes in thermal environment can invoke a stress response in pigs, which in turn can potentially impact their immune system and disease susceptibility. We investigated effects of heat stress and social rank on behavior, immune responsiveness, and performance of pigs challenged with porcine reproductive and respiratory syndrome (PRRS) virus. Sixty-four 7-week-old PRRS-naïve pigs were assigned to each of four experimental treatments consisting of a 2 x 2 factorial design: PRRS (PRRS- or PRRS+) and environmental temperature (24 degrees C or 32 degrees C).

Photoperiod influences the immune status of multiparous pregnant sows and their piglets.

Manipulation of photoperiod may provide a noninvasive, easily implemented, effective method to improve immune status and enhance the efficiency of production. The objective of this study was to evaluate the impact of manipulation of photoperiod on endocrine and immune responses of pregnant sows and their offspring. At d 83 of gestation, sows were moved to gestation stalls and kept on a photoperiod of 12 h of light:12 h of dark until d 90, when sows were assigned to a long day (LD; 16 h of light/d) or a short day (SD; 8 h of light/d) treatment.

Impacts of chronic stress and social status on various physiological and performance measures in pigs of different breeds.

Pigs typically experience various environmental stressors, which can negatively affect performance. Cortisol concentrations and various immune and performance measures are influenced by breed, but few data exist describing the impact of breed on stress responsiveness in pigs. The objective of this experiment was to determine if certain physiological responses to chronic stressors differed among 3 breeds and 2 commercial lines of pigs.