The fork protection complex generates DNA topological stress-induced DNA damage while ensuring full and faithful genome duplication.

The fork protection complex (FPC), composed of Mrc1, Tof1, and Csm3, supports rapid and stable DNA replication. Here, we show that FPC activity also introduces DNA damage by increasing DNA topological stress during replication. Mrc1 action increases DNA topological stress during plasmid replication, while Mrc1 or Tof1 activity causes replication stress and DNA damage within topologically constrained regions.

To precept or not to precept: Perspectives from nurse practitioners.

Background: Nurse practitioner (NP) preceptors are crucial to clinical education. Recent increases in the required direct patient care hours for NP students may worsen the current preceptor shortage. However, most studies of preceptorship only include NPs who are current preceptors and are therefore missing vital information from nonpreceptors.

Entangling and disentangling mitotic chromosomes.

The DNA topological challenges generated by cellular manipulation of extremely long DNA fibers remain poorly understood. In this issue of Molecular Cell, Hildebrand et al. describe how mitotic chromosomes are self entangled and that disentanglement requires TOP2 activity in late mitosis.

Sister chromatid cohesion establishment during DNA replication termination.

Newly copied sister chromatids are tethered together by the cohesin complex, but how sister chromatid cohesion coordinates with DNA replication is poorly understood. Prevailing models suggest that cohesin complexes, bound to DNA before replication, remain behind the advancing replication fork to keep sister chromatids together. By visualizing single replication forks colliding with preloaded cohesin complexes, we find that the replisome instead pushes cohesin to where a converging replisome is met.

The Validation of Surgical Simulators: A Technical Report on Current Validation Terminology as a Reference for Future Research.

In recent years, surgical trainees have been exposed to a lower volume of operative procedures. In part, this is due to the reduction in working hours and further disruption by the coronavirus disease 2019 pandemic. Much has been done to develop the techniques of surgical skill training outside of the operating theatre.

Cryo-EM structure of the Smc5/6 holo-complex.

The Smc5/6 complex plays an essential role in the resolution of recombination intermediates formed during mitosis or meiosis, or as a result of the cellular response to replication stress. It also functions as a restriction factor preventing viral replication. Here, we report the cryogenic EM (cryo-EM) structure of the six-subunit budding yeast Smc5/6 holo-complex, reconstituted from recombinant proteins expressed in insect cells - providing both an architectural overview of the entire complex and an understanding of how the Nse1/3/4 subcomplex binds to the hetero-dimeric SMC protein core.

Telomerase subunit Est2 marks internal sites that are prone to accumulate DNA damage.

Background: The main function of telomerase is at the telomeres but under adverse conditions telomerase can bind to internal regions causing deleterious effects as observed in cancer cells.

Results: By mapping the global occupancy of the catalytic subunit of telomerase (Est2) in the budding yeast Saccharomyces cerevisiae, we reveal that it binds to multiple guanine-rich genomic loci, which we termed "non-telomeric binding sites" (NTBS). We characterize Est2 binding to NTBS.

The FAST Workstation Shows Construct Validity and Participant Endorsement.

Purpose: To determine in what way the proposed simulation-based intervention (SBI) is an effective intervention for use in basic arthroscopic skills training.

Methods: Twenty candidates were recruited and grouped according to experience. Performance metrics included the time to activity completion, errors made, and Global Rating Scale score.

Elasticity of the coracohumeral ligament in patients with frozen shoulder following rotator interval injection: a case series.

To evaluate changes in the elasticity of the coracohumeral ligament in patients with adhesive capsulitis of the shoulder treated with ultrasound-guided rotator interval injections. Shear wave elastography was used to evaluate elasticity of the coracohumeral ligament in symptomatic and asymptomatic shoulders in the shoulder-neutral position and 30° external rotation. A total of 24 shoulders were assessed.

Separable functions of Tof1/Timeless in intra-S-checkpoint signalling, replisome stability and DNA topological stress.

The highly conserved Tof1/Timeless proteins minimise replication stress and promote normal DNA replication. They are required to mediate the DNA replication checkpoint (DRC), the stable pausing of forks at protein fork blocks, the coupling of DNA helicase and polymerase functions during replication stress (RS) and the preferential resolution of DNA topological stress ahead of the fork. Here we demonstrate that the roles of the Saccharomyces cerevisiae Timeless protein Tof1 in DRC signalling and resolution of DNA topological stress require distinct N and C terminal regions of the protein, whereas the other functions of Tof1 are closely linked to the stable interaction between Tof1 and its constitutive binding partner Csm3/Tipin.

A systematic review of pre-hospital shoulder reduction techniques for anterior shoulder dislocation and the effect on patient return to function.

Purpose: The majority of acute anterior shoulder dislocations are sustained during sports and wilderness activities. The management of acute dislocations in the pre-hospital setting is currently without guidelines based on the evidence. The study aims to assess the risk of acute complications in pre-hospital shoulder reduction and identify which pre-hospital reduction technique has the highest success rate in the published literature.

Convergent genes shape budding yeast pericentromeres.

The three-dimensional architecture of the genome governs its maintenance, expression and transmission. The cohesin protein complex organizes the genome by topologically linking distant loci, and is highly enriched in specialized chromosomal domains surrounding centromeres, called pericentromeres. Here we report the three-dimensional structure of pericentromeres in budding yeast (Saccharomyces cerevisiae) and establish the relationship between genome organization and function.

Cohesin Causes Replicative DNA Damage by Trapping DNA Topological Stress.

DNA topological stress inhibits DNA replication fork (RF) progression and contributes to DNA replication stress. In Saccharomyces cerevisiae, we demonstrate that centromeric DNA and the rDNA array are especially vulnerable to DNA topological stress during replication. The activity of the SMC complexes cohesin and condensin are linked to both the generation and repair of DNA topological-stress-linked damage in these regions.

Principles of meiotic chromosome assembly revealed in S. cerevisiae.

During meiotic prophase, chromosomes organise into a series of chromatin loops emanating from a proteinaceous axis, but the mechanisms of assembly remain unclear. Here we use Saccharomyces cerevisiae to explore how this elaborate three-dimensional chromosome organisation is linked to genomic sequence. As cells enter meiosis, we observe that strong cohesin-dependent grid-like Hi-C interaction patterns emerge, reminiscent of mammalian interphase organisation, but with distinct regulation.

Checkpoint inhibition of origin firing prevents DNA topological stress.

A universal feature of DNA damage and replication stress in eukaryotes is the activation of a checkpoint-kinase response. In S-phase, the checkpoint inhibits replication initiation, yet the function of this global block to origin firing remains unknown. To establish the physiological roles of this arm of the checkpoint, we analyzed separation of function mutants in the budding yeast that allow global origin firing upon replication stress, despite an otherwise normal checkpoint response.

Preparation of Cell Cycle-Synchronized Saccharomyces cerevisiae Cells for Hi-C.

The chromosome organization activities of SMC complexes are crucial for correct gene expression and genetic inheritance in cells. Hi-C assays have revealed previously unsuspected levels of chromosome structure, with different types of chromosome structure facilitating function at different stages of the cell cycle. Elucidating how SMC complexes regulate these distinct types of organization is currently a key question in molecular biology.

Pif1-Family Helicases Support Fork Convergence during DNA Replication Termination in Eukaryotes.

The convergence of two DNA replication forks creates unique problems during DNA replication termination. In E. coli and SV40, the release of torsional strain by type II topoisomerases is critical for converging replisomes to complete DNA synthesis, but the pathways that mediate fork convergence in eukaryotes are unknown.

Are SMC Complexes Loop Extruding Factors? Linking Theory With Fact.

The extreme length of chromosomal DNA requires organizing mechanisms to both promote functional genetic interactions and ensure faithful chromosome segregation when cells divide. Microscopy and genome-wide contact frequency analyses indicate that intra-chromosomal looping of DNA is a primary pathway of chromosomal organization during all stages of the cell cycle. DNA loop extrusion has emerged as a unifying model for how chromosome loops are formed in cis in different genomic contexts and cell cycle stages.

Functional outcome of open acromioclavicular joint stabilization for instability following distal clavicle resection.

Background: Acromioclavicular joint instability following distal clavicle resection can result in considerable pain and dysfunction.

Method: We present a review of 13 patients who underwent ACJ stabilization following one or more distal clavicle resection procedures.

Results: The mean Quick DASH and CM scores were 26(0-57) and 73(46-100) respectively.

SMC complexes differentially compact mitotic chromosomes according to genomic context.

Structural maintenance of chromosomes (SMC) protein complexes are key determinants of chromosome conformation. Using Hi-C and polymer modelling, we study how cohesin and condensin, two deeply conserved SMC complexes, organize chromosomes in the budding yeast Saccharomyces cerevisiae. The canonical role of cohesin is to co-align sister chromatids, while condensin generally compacts mitotic chromosomes.

Viscoelastic properties of mitral valve leaflets: An analysis of regional variation and frequency-dependency.

The aim of this study was to determine the regional variation in viscoelastic properties of mitral valve leaflets over a range of physiological and patho-physiological frequencies. This included comparisons to be made between anterior and posterior leaflets, anterior leaflet clear and rough zones, and radial and circumferential leaflet orientation. Dynamic mechanical analysis was used to determine frequency-dependent viscoelastic properties.

The Causes and Consequences of Topological Stress during DNA Replication.

The faithful replication of sister chromatids is essential for genomic integrity in every cell division. The replication machinery must overcome numerous difficulties in every round of replication, including DNA topological stress. Topological stress arises due to the double-stranded helical nature of DNA.