Establishment of a replication fork barrier following induction of DNA binding in mammalian cells

Understanding the mechanisms that lead to replication fork blocks (RFB) and the means to bypass them is important given the threat that they represent for genome stability if inappropriately handled. Here, to study this issue in mammals, we use integrated arrays of the LacO and/or TetO as a tractable system to follow in time a process in an individual cell and at a single locus. Importantly, we show that induction of the binding by LacI and TetR proteins, and not the presence of the repeats, is key to form the RFB. We find that the binding of the proteins to the arrays during replication causes a prolonged persistence of replication foci at the site. This, in turn, induces a local DNA damage repair (DDR) response, with the recruitment of proteins involved in double-strand break (DSB) repair such as TOPBP1 and 53BP1, and the phosphorylation of H2AX. Furthermore, the appearance of micronuclei and DNA bridges after mitosis is consistent with an incomplete replication. We discuss how the many DNA binding proteins encountered during replication can be dealt with and the consequences of incomplete replication. Future studies exploiting this type of system should help analyze how an RFB, along with bypass mechanisms, are controlled in order to maintain genome integrity.     

A new Middle Devonian gasterocomid crinoid from central Turkey and revision of the Gasterocomidae

A juvenile cup, disarticulated arm plates, and columnals of the crinoid Arachnocrinus sarizensis n. sp. are described from the Eastern Taurus Mountains of central Turkey. This is the first Paleozoic crinoid based on a cup and arm plates reported from Turkey. Suggested revision of the Gasterocomidae includes transfer of Arachnocrinus and Ancyrocrinus to the family and rejection of Kopficrinus from the family. The stratigraphic range of Arachnocrinus is extended upward into the Middle Devonian from the Early Devonian. The Devonian of Turkey is of special interest because it includes Laurasian and Gondwanan components on different tectonic blocks. The paleogeographic range of Arachnocrinus is extended from the North American plate onto the Anatolian block, which would have been located on the southern edge of the Paleotethys Sea in the Middle Devonian in a shelf basin off the northern coast of Gondwana at approximately 42° south latitude.     

Ichnological and Sedimentological Characteristics of Submarine Fan-Delta Deposits in a Half-Graben,Lower Cretaceous Palnatokes Bjerg Formation,NE Greenland

Fault-scarp related fan deltas developed in tilted half grabens in NE Greenland during late Jurassic—early Cretaceous rifting. This study documents ichnological and sedimentological characteristics of the Lower Cretaceous interval of a submarine fan-delta succession (Palnatokes Bjerg Formation, Wollaston Forland), which represents a time of waning rift activity and transgression. For this purpose, two variably exposed ca. 150 m-thick sections were studied ～10 km from the coeval fault scarp, near the axis of the most proximal fault block. Moreover, an additional ～20 m thick coeval succession was studied in the next fault block ～20 km from the coastline defining fault. The results indicate deposition on the basin floor, in the distal fan and in a mid-fan channel-overbank/splay complex of a subaqueous fan delta. The deposits are characterized mainly by various facies of high and low density turbidity currents, hybrid event beds, and transitional flow facies that grade upward into sediment starved basinal mudstones. The ichnological pattern recorded in these strata is strongly mixed, frequently containing elements of the impoverished Skolithos, Cruziana, Zoophycos ichnofacies, and more rarely of the Nereites ichnofacies. Characteristic features also include suites dominated by infaunal locomotion and feeding trails (including the “Curvolithus suite”) and the common occurrence of crustacean burrows. The results are indicative of a depositional system resembling a fjord-side delta that differs sedimentologically and ichnologically from many other gravity-flow systems of similar grain-size caliber. The ichnological pattern recorded in these strata is potentially a characteristic feature of the subaqueous fan-deltas in comparable settings, reflecting the distinct basin physiography with an abrupt change in bathymetry, a narrow basin geometry, and environmental stress resulting from unstable physical conditions. The counter slope of the rotated fault block may explain the common signs of flow concentration and abrupt fan termination.     

Improvements in the determination of radionuclide distribution in the analysis of irradiated samples using double-differentiation method

The spatial response of an 8x4 block detector made up of 5.6-mm-wide, 12.9-mm-high, 30-mm-thick individual detector crystals to a collimated line source of 511 keV annihilation photons was examined. The response of each crystal showed a spread around the average positioning values and distributions from adjacent crystals overlapped as the collimated source scanned the individual detectors. This leads to possible errors in the event assignment. The implementation of double differentiation or the second derivative method was proposed for the removal of scattered photons so as to reduce the overlap and, hence, avoid mispositioning. This method is a mathematical solution implemented when analysing the results. A curve in a spatial spectrum could be considered to be a function f(x), wherex is the position. When double differentiation of f(x) is carried out, then the normalized curve d²f(x) appears with some reduction in the wings. It was shown that a reduction of the scattering contribution in the tails without overestimating the contribution of scattered events could be achieved by implementing a double-differentiation process.     

2021 PACES expert consensus statement on the indications and management of cardiovascular implantable electronic devices in pediatric patients

In view of the increasing complexity of both cardiovascular implantable electronic devices (CIEDs) and patients in the current era, practice guidelines, by necessity, have become increasingly specific. This document is an expert consensus statement that has been developed to update and further delineate indications and management of CIEDs in pediatric patients, defined as ≤21 years of age, and is intended to focus primarily on the indications for CIEDs in the setting of specific disease categories. The document also highlights variations between previously published adult and pediatric CIED recommendations and provides rationale for underlying important differences. The document addresses some of the deterrents to CIED access in low- and middle-income countries and strategies to circumvent them. The document sections were divided up and drafted by the writing committee members according to their expertise. The recommendations represent the consensus opinion of the entire writing committee, graded by class of recommendation and level of evidence. Several questions addressed in this document either do not lend themselves to clinical trials or are rare disease entities, and in these instances recommendations are based on consensus expert opinion. Furthermore, specific recommendations, even when supported by substantial data, do not replace the need for clinical judgment and patient-specific decision-making. The recommendations were opened for public comment to Pediatric and Congenital Electrophysiology Society (PACES) members and underwent external review by the scientific and clinical document committee of the Heart Rhythm Society (HRS), the science advisory and coordinating committee of the American Heart Association (AHA), the American College of Cardiology (ACC), and the Association for European Paediatric and Congenital Cardiology (AEPC). The document received endorsement by all the collaborators and the Asia Pacific Heart Rhythm Society (APHRS), the Indian Heart Rhythm Society (IHRS), and the Latin American Heart Rhythm Society (LAHRS). This document is expected to provide support for clinicians and patients to allow for appropriate CIED use, appropriate CIED management, and appropriate CIED follow-up in pediatric patients.     

Rank Tests for correlated Random Variables

In this paper we consider the asymptotic distribution of linear rank-statistics allowing certain dependencies between the random variables. General theorems will be given, from which the results contained in SEN (1967a, 1967b, 1968), MEHRA /SEN (1969) and PURI /SEN (1971) can be derived. Applying these theorems we can derive ranktests for the two-sample problem for tied observations, for the two-factor mixed model with random interaction and for a hierarchical design. The distribution of the proposed teststatistics for small sample sizes is considered by Monte-Carlo studies.     

Xyloglucan-based diblock co-oligomer: Synthesis,self-assembly and steric stabilization of proteins

We propose a novel plant-based amphiphilic diblock co-oligomers (BCO) surfactant containing only carbohydrate segments and examine its potential as a biosourced stabilizer. The synthesis of an amphiphilic xyloglucan-based BCO, composed of a hydrophilic xyloglucan oligosaccharide (XGO) block “clicked” to a hydrophobic peracetylated XGO is described. Dynamic light scattering experiments correlated with transmission electron microscopy observations showed that this new class of amphiphilic BCO self-assembles in water to form spherical micelles with a hydrodynamic diameter of 22 nm. Preliminary studies indicate that the XGO-based BCO sterically stabilizes gliadin and zein nanoparticle suspensions. The stabilization results were compared to those using pluronic F-68, a commercial surfactant. For gliadin nanoparticles, both surfactants result in essentially the same morphology and polydispersity. However, for the zein nanoparticles, the XGO-based BCO stabilizer gave lower polydispersity.     

Optimization of the alkaline comet assay for easy repair capacity quantification of oxidative DNA damage in PBMC from human volunteers using aphidicolin block

Assessment of DNA repair capacity (DRC) upon ex vivo challenge of peripheral blood mononuclear cells (PBMC) with oxidative damage inducing agents, as evaluated by the comet assay, is widely used as biomarker to assess the antioxidant status in human studies. Here, the alkaline comet assay was now optimized for easy and time saving detection of repair capacity upon oxidative stress-induced DNA damage using the DNA polymerase inhibitor aphidicolin (APC) to block repair of hydrogen peroxide (H₂O₂) induced DNA damage. Addition of a DMSO-containing DNA damage stop solution was found suitable to replace washing steps for H₂O₂ removal before APC block. Cell treatment with APC at 6 μM did not impact baseline DNA damage but could reliably block DNA repair after H₂O₂ challenge in both fresh and cryopreserved samples thus omitting the use of a starting time point control. Under the conditions used, frozen cells, with or without an additional 4 h rest, showed the same repair capacity as their fresh counterpart. The intra assay coefficient of variation (CV) was 3.3%. To provide proof of principle, the modified assay was applied to cryopreserved PBMC from 19 participants of a short-term Brassica diet intervention study investigating potential health promoting effects of the food intervention. Then, a 33% increase in DRC (p ≤ 0.01) could be shown in samples after intervention (mean ± SD: 5.82 ± 1) as compared to baseline (mean ± SD: 4.38 ± 1.21). Individual samples from baseline and intervention showed an inter-individual CV of 27.65% (baseline) and 17.26% (intervention). Taken together this modified comet assay protocol allows the facilitated detection of DNA repair in fresh or cryopreserved human PBMC samples with a good sensitivity and reliability and could be useful in human studies addressing the antioxidant status and repair capacity of PBMC.