Xenopus Drf1, a Regulator of Cdc7, Displays Checkpoint-dependent Accumulation on Chromatin during an S-phase Arrest

We have cloned a Xenopus Dbf4-related factor named Drf1 and characterized this protein by using Xenopus egg extracts. Drf1 forms an active complex with the kinase Cdc7. However, most of the Cdc7 in egg extracts is not associated with Drf1, which raises the possibility that some or all of the remaining Cdc7 is bound to another Dbf4-related protein. Immunodepletion of Drf1 does not prevent DNA replication in egg extracts. Consistent with this observation, Cdc45 can still associate with chromatin in Drf1-depleted extracts, albeit at significantly reduced levels. Nonetheless, Drf1 displays highly regulated binding to replicating chromatin. Treatment of egg extracts with aphidicolin results in a substantial accumulation of Drf1 on chromatin. This accumulation is blocked by addition of caffeine and by immunodepletion of either ATR or Claspin. These observations suggest that the increased binding of Drf1 to aphidicolin-treated chromatin is an active process that is mediated by a caffeine-sensitive checkpoint pathway containing ATR and Claspin. Abrogation of this pathway also leads to a large increase in the binding of Cdc45 to chromatin. This increase is substantially reduced in the absence of Drf1, which suggests that regulation of Drf1 might be involved in the suppression of Cdc45 loading during replication arrest. We also provide evidence that elimination of this checkpoint causes resumed initiation of DNA replication in both Xenopus tissue culture cells and egg extracts. Taken together, these observations argue that Drf1 is regulated by an intra-S-phase checkpoint mechanism that down-regulates the loading of Cdc45 onto chromatin containing DNA replication blocks

Spectroscopy and Imaging Performance of the Liquid Xenon Gamma-Ray Imaging Telescope (LXeGRIT)

LXeGRIT is a balloon-borne Compton telescope based on a liquid xenon time projection chamber (LXeTPC) for imaging cosmic \g-rays in the energy band of 0.2-20 MeV. The detector, with 400 cm$^2$ area and 7 cm drift gap, is filled with high purity LXe. Both ionization and scintillation light signals are detected to measure the energy deposits and the three spatial coordinates of individual \g -ray interactions within the sensitive volume. The TPC has been characterized with repeated measurements of its spectral and Compton imaging response to \g -rays from radioactive sources such as \na, \cs, \yt and Am-Be. The detector shows a linear response to \g -rays in the energy range 511 keV -4.4 MeV, with an energy resolution (FWHM) of \Delta E/E=8.8% \: \sqrt{1\MeV /E}. Compton imaging of \yt \g -ray events with two detected interactions is consistent with an angular resolution of $\sim$ 3 degrees (RMS) at 1.8 MeV.Comment: To appear in: Hard X-Ray, Gamma-Ray and Neutron Detector Physics XI, 2000; Proc. SPIE, vol. 4140; K.A. Flanagan & O.H. Siegmund, ed

Feasibility randomised controlled trial comparing TRAK-ACL digital rehabilitation intervention plus treatment as usual versus treatment as usual for patients following anterior cruciate ligament reconstruction

Objectives: To evaluate the feasibility of trialling taxonomy for the rehabilitation of knee conditions-ACL (TRAK-ACL), a digital health intervention that provides health information, personalised exercise plans and remote clinical support combined with treatment as usual (TAU), for people following ACL reconstruction. Methods: The study design was a two-arm parallel randomised controlled trial (RCT). Eligible participants were English-speaking adults who had undergone ACL reconstruction within the last 12 weeks, had access to the internet and could provide informed consent. Recruitment took place at three sites in the UK. TRAK-ACL intervention was an interactive website informed by behaviour change technique combined with TAU. The comparator was TAU. Outcomes were: recruitment and retention; completeness of outcome measures at follow-up; fidelity of intervention delivery and engagement with the intervention. Individuals were randomised using a computer-generated random number sequence. Blinded assessors allocated groups and collected outcome measures. Results: Fifty-nine people were assessed for eligibility at two of the participating sites, and 51 were randomised; 26 were allocated to TRAK-ACL and 25 to TAU. Follow-up data were collected on 44 and 40 participants at 3 and 6 months, respectively. All outcome measures were completed fully at 6 months except the Client Service Receipt Inventory. Two patients in each arm did not receive the treatment they were randomised to. Engagement with TRAK-ACL intervention was a median of 5 logins (IQR 3-13 logins), over 18 weeks (SD 12.2 weeks). Conclusion: TRAK-ACL would be suitable for evaluation of effectiveness in a fully powered RCT

Metabolism of tissue macrophages in homeostasis and pathology.

Cellular metabolism orchestrates the intricate use of tissue fuels for catabolism and anabolism to generate cellular energy and structural components. The emerging field of immunometabolism highlights the importance of cellular metabolism for the maintenance and activities of immune cells. Macrophages are embryo- or adult bone marrow-derived leukocytes that are key for healthy tissue homeostasis but can also contribute to pathologies such as metabolic syndrome, atherosclerosis, fibrosis or cancer. Macrophage metabolism has largely been studied in vitro. However, different organs contain diverse macrophage populations that specialize in distinct and often tissue-specific functions. This context specificity creates diverging metabolic challenges for tissue macrophage populations to fulfill their homeostatic roles in their particular microenvironment and conditions their response in pathological conditions. Here, we outline current knowledge on the metabolic requirements and adaptations of macrophages located in tissues during homeostasis and selected diseases.SKW and the project that gave rise to these results received support in the form of a fellowship from the La Caixa Foundation (ID 100010434). The fellowship code is LCF/BQ/ PR20/11770008. GD is supported by a European Molecular Biology Organization Longterm Fellowship (ALTF 379-2019). This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie SkłodowskaCurie grant agreement No. 892965. IHM is supported by a La Caixa INPhINIT fellowship (ID 100010434, fellowship code: LCF/BQ/IN17/11620074). Work in the DS laboratory is funded by the CNIC, by the European Research Council (ERC-2016-Consolidator Grant 725091), by the Agencia Estatal de Investigación (PID2019-108157RB), by the Comunidad de Madrid (B2017/BMD-3733 Immunothercan-CM), by Atresmedia (Constantes y Vitales prize), by the Fondo Solidario Juntos (Banco Santander), and by the Fundació La Marató de TV3 (201723). The CNIC is supported by the Instituto de Salud Carlos III (ISCIII), the MICINN and the Pro CNIC Foundation.S

The Arf6 GEF GEP100/BRAG2 Regulates Cell Adhesion by Controlling Endocytosis of β1 Integrins

SummaryThe small GTPase Arf6 has been shown to regulate the post-endocytic trafficking of a subset of membrane proteins, including β1 integrins, and inhibition of Arf6 function impairs both cell adhesion and motility [1]. The activity of Arf GTPases is regulated by a large family of guanine nucleotide exchange factors (GEFs) [2]. Arf-GEP100/BRAG2 is a GEF with reported specificity for Arf6 in vitro [3], but it is otherwise poorly characterized. Here we report that BRAG2 exists in two ubiquitously expressed isoforms, which we call BRAG2a and BRAG2b, both of which can activate Arf6 in vivo. Depletion of endogenous BRAG2 by siRNA leads to dramatic effects in the cell periphery; one such effect is an accumulation of β1 integrin on the cell surface and a corresponding enhancement of cell attachment and spreading on fibronectin-coated substrates. In contrast, depletion of Arf6 leads to intracellular accumulation of β1 integrin and reduced adhesion and spreading. These findings suggest that Arf6 regulates both endocytosis and recycling of β1 integrins and that BRAG2 functions selectively to activate Arf6 during integrin internalization

THE ROLE OF ANXIETY IN GOLF PUTTING PERFORMANCE

Anxiety’s influence on performance continues to be one of the main research interests for sport psychologists (Hanin, 2000). It is apparent, though, that there is a lack of empirical research characterising the multi-disciplinary effect of anxiety on sports performance. The current study aimed to ascertain biomechanical (accuracy, movement variability) and psychological (anxiety) markers to determine how anxiety affects golf putting

MiR203 Mediates Subversion of Stem Cell Properties During Mammary Epithelial Differentiation via Repression of ΔNP63α and Promotes Mesenchymal-to-Epithelial Transition

During reproductive life, the mammary epithelium undergoes consecutive cycles of proliferation, differentiation and apoptosis. Doing so relies on the retained proliferative capacity, prolonged lifespan and developmental potency of mammary stem cells (MaSCs). ΔNp63α, the predominant TP63 isoform in mammary epithelia, is robustly expressed in MaSCs and is required for preservation of self-renewing capacity in diverse epithelial structures. However, the mechanism(s) underlying subversion of this activity during forfeiture of self-renewing capacity are poorly understood. MicroRNAs (miRNAs) govern critical cellular functions including stem cell maintenance, development, cell cycle regulation and differentiation by disrupting translation of target mRNAs. Data presented here indicate that expression of miR203, a miRNA that targets ΔNp63α and ΔNp63β is activated during luminal epithelial differentiation and that this pattern is observed in the murine mammary hierarchy. In addition, we present evidence that the transcription factor Zeb1 represses miR203 expression, thus enhancing ΔNp63α protein levels. Furthermore, ectopic miR203 suppresses ΔNp63α expression, proliferation and colony formation. The anti-clonogenic effects mediated by miR203 require suppression of ΔNp63α. In addition, ectopic miR203 promotes mesenchymal-to-epithelial transition and disrupts activities associated with epithelial stem cells. These studies support a model in which induction of miR203 mediates forfeiture of self-renewing capacity via suppression of ΔNp63α and may also have anti-tumorigenic activity through its reduction of EMT and cancer stem cell populations

ASSESSING AND REDUCING SOYBEAN CROP LOSSES FROM DEER: AN INTERDISCIPLINARY, MULTI-AGENCY EFFORT

Damage from white-tailed deer (Odocoileus virginianus) has become a common complaint of soybean (Glycine max) producers in many areas of the Southeast. Both short- and long-term, single-field and community-wide solutions to this problem are needed. This paper describes a multi-agency, multi-state effort, involving agronomists, wildlife biologists, producers, and other landowners, to assess soybean losses from deer and to evaluate potential solutions. One phase of this work, which is supported by soybean producer checkoff funds, involves evaluating agronomic practices for reducing crop losses. These include drilled (rather than wide-row) plantings and use of insect-resistant or dense-pubescent cultivars (varieties) which may deter browsing, especially where deer pressure is light to moderate. Evaluations of these practices, in comparison with conventional ones, are being conducted in producer’s fields in SC, NC, and VA. The other phase of this work is a cooperative project involving Clemson University, the SC Wildlife and Marine Resources Department, soybean producers and’ other landowners in a 7500-acre tract in Hampton and Jasper Cos., SC. The deer population in this tract will be monitored and reduced over a 3-year period, and the resulting effects on soybean crop losses and herd quality will be assessed