Saccharomyces cerevisiae Mre11 is a high-affinity G4 DNA-binding protein and a G-rich DNA-specific endonuclease: implications for replication of telomeric DNA

In Saccharomyces cerevisiae, Mre11p/Rad50p/Xrs2p (MRX) complex plays a vital role in several nuclear processes including cellular response to DNA damage, telomere length maintenance, cell cycle checkpoint control and meiotic recombination. Telomeres are comprised of tandem repeats of G-rich DNA and are incorporated into non-nucleosomal chromatin. Although the structure of the yeast telomeric DNA is poorly understood, it has been suggested that the G-rich sequences can fold into G4 DNA, which has been shown to inhibit DNA synthesis by telomerase. However, little is known about the factors and mechanistic aspects of the generation of appropriate termini for DNA synthesis by telomerase. Here, we show that S.cerevisiae Mre11 protein (ScMre11p) possesses substantially higher binding affinity for G4 DNA, over single- or double-stranded DNA, and binding was inhibited by poly(dG) or porphyrin. Binding of ScMre11p to G4 DNA was most robust, compared with G2′ DNA and the resulting protein–DNA complexes were strikingly very resistant to dissociation by NaCl. Remarkably, binding of ScMre11p to G4 DNA and G-rich single-stranded DNA was accompanied by the endonucleolytic cleavage at sites flanking the array of G residues and G-quartets in Mn(2+)-dependent manner. Collectively, these results suggest that ScMre11p is likely to play a major role in generating appropriate substrates for DNA synthesis by telomerase and telomere-binding proteins. We discuss the implications of these findings with regard to telomere length maintenance by telomerase-dependent and independent mechanisms

Quantum Gas Mixtures and Dual-Species Atom Interferometry in Space

The capability to reach ultracold atomic temperatures in compact instruments has recently been extended into space. Ultracold temperatures amplify quantum effects, while free-fall allows further cooling and longer interactions time with gravity - the final force without a quantum description. On Earth, these devices have produced macroscopic quantum phenomena such as Bose-Einstein condensation (BECs), superfluidity, and strongly interacting quantum gases. Quantum sensors interfering the superposition of two ultracold atomic isotopes have tested the Universality of Free Fall (UFF), a core tenet of Einstein's classical gravitational theory, at the $10^{-12}$ level. In space, cooling the elements needed to explore the rich physics of strong interactions and preparing the multiple species required for quantum tests of the UFF has remained elusive. Here, utilizing upgraded capabilities of the multi-user Cold Atom Lab (CAL) instrument within the International Space Station (ISS), we report the first simultaneous production of a dual species Bose-Einstein condensate in space (formed from $^{87}$Rb and $^{41}$K), observation of interspecies interactions, as well as the production of $^{39}$K ultracold gases. We have further achieved the first space-borne demonstration of simultaneous atom interferometry with two atomic species ($^{87}$Rb and $^{41}$K). These results are an important step towards quantum tests of UFF in space, and will allow scientists to investigate aspects of few-body physics, quantum chemistry, and fundamental physics in novel regimes without the perturbing asymmetry of gravity

Short Telomeres Initiate Telomere Recombination in Primary and Tumor Cells

Human tumors that lack telomerase maintain telomeres by alternative lengthening mechanisms. Tumors can also form in telomerase-deficient mice; however, the genetic mechanism responsible for tumor growth without telomerase is unknown. In yeast, several different recombination pathways maintain telomeres in the absence of telomerase—some result in telomere maintenance with minimal effects on telomere length. To examine non-telomerase mechanisms for telomere maintenance in mammalian cells, we used primary cells and lymphomas from telomerase-deficient mice (mTR−/− and Eμmyc+mTR−/−) and CAST/EiJ mouse embryonic fibroblast cells. These cells were analyzed using pq-ratio analysis, telomere length distribution outliers, CO-FISH, Q-FISH, and multicolor FISH to detect subtelomeric recombination. Telomere length was maintained during long-term growth in vivo and in vitro. Long telomeres, characteristic of human ALT cells, were not observed in either late passage or mTR−/− tumor cells; instead, we observed only minimal changes in telomere length. Telomere length variation and subtelomeric recombination were frequent in cells with short telomeres, indicating that length maintenance is due to telomeric recombination. We also detected telomere length changes in primary mTR−/− cells that had short telomeres. Using mouse mTR+/− and human hTERT+/− primary cells with short telomeres, we found frequent length changes indicative of recombination. We conclude that telomere maintenance by non-telomerase mechanisms, including recombination, occurs in primary cells and is initiated by short telomeres, even in the presence of telomerase. Most intriguing, our data indicate that some non-telomerase telomere maintenance mechanisms occur without a significant increase in telomere length

Site of Allergic Airway Narrowing and the Influence of Exogenous Surfactant in the Brown Norway Rat

Background: The parameters RN (Newtonian resistance), G (tissue damping), and H (tissue elastance) of the constant phase model of respiratory mechanics provide information concerning the site of altered mechanical properties of the lung. The aims of this study were to compare the site of allergic airway narrowing implied from respiratory mechanics to a direct assessment by morphometry and to evaluate the effects of exogenous surfactant administration on the site and magnitude of airway narrowing. Methods: We induced airway narrowing by ovalbumin sensitization and challenge and we tested the effects of a natural surfactant lacking surfactant proteins A and D (InfasurfH) on airway responses. Sensitized, mechanically ventilated Brown Norway rats underwent an aerosol challenge with 5 % ovalbumin or vehicle. Other animals received nebulized surfactant prior to challenge. Three or 20 minutes after ovalbumin challenge, airway luminal areas were assessed on snap-frozen lungs by morphometry. Results: At 3 minutes, RN and G detected large airway narrowing whereas at 20 minutes G and H detected small airway narrowing. Surfactant inhibited RN at the peak of the early allergic response and ovalbumin-induced increase in bronchoalveolar lavage fluid cysteinyl leukotrienes and amphiregulin but not IgE-induced mast cell activation in vitro. Conclusion: Allergen challenge triggers the rapid onset of large airway narrowing, detected by RN and G, and subsequen

National laboratory-based surveillance system for antimicrobial resistance: a successful tool to support the control of antimicrobial resistance in the Netherlands

An important cornerstone in the control of antimicrobial resistance (AMR) is a well-designed quantitative system for the surveillance of spread and temporal trends in AMR. Since 2008, the Dutch national AMR surveillance system, based on routine data from medical microbiological laboratories (MMLs), has developed into a successful tool to support the control of AMR in the Netherlands. It provides background information for policy making in public health and healthcare services, supports development of empirical antibiotic therapy guidelines and facilitates in-depth research. In addition, participation of the MMLs in the national AMR surveillance network has contributed to sharing of knowledge and quality improvement. A future improvement will be the implementation of a new semantic standard together with standardised data transfer, which will reduce errors in data handling and enable a more real-time surveillance. Furthermore, the

Post translational changes to α-synuclein control iron and dopamine trafficking : a concept for neuron vulnerability in Parkinson's disease

Parkinson's disease is a multifactorial neurodegenerative disorder, the aetiology of which remains elusive. The primary clinical feature of progressively impaired motor control is caused by a loss of midbrain substantia nigra dopamine neurons that have a high α-synuclein (α-syn) and iron content. α-Syn is a neuronal protein that is highly modified post-translationally and central to the Lewy body neuropathology of the disease. This review provides an overview of findings on the role post translational modifications to α-syn have in membrane binding and intracellular vesicle trafficking. Furthermore, we propose a concept in which acetylation and phosphorylation of α-syn modulate endocytic import of iron and vesicle transport of dopamine during normal physiology. Disregulated phosphorylation and oxidation of α-syn mediate iron and dopamine dependent oxidative stress through impaired cellular location and increase propensity for α-syn aggregation. The proposition highlights a connection between α-syn, iron and dopamine, three pathological components associated with disease progression in sporadic Parkinson's disease

Habitat preferences by individual humpback whale mothers in the Hawaiian breeding grounds vary with the age and size of their calves

We investigated whether calf age and calf size influence habitat choice by humpback whale motherecalf pairs in their breeding grounds. During 1997e2008, we conducted focal follows of motherecalf pairs in Hawaiian waters. Tail-fluke identification photographs and calf lengths (measured through video-grammetry) were obtained. Water depth and sea-bed terrain type were derived from GPS data. Identification photographs were matched so that the habitat choices could be established within breeding seasons. Across 72 motherecalf pairs resighted over various intervals within a breeding season, magnitude of depth change between initial and final sightings increased significantly with resighting interval. There was a significant increase from initial depth to final depth for relatively long resighting intervals (27e51 days), but no significant difference for relatively short resighting intervals (2e26 days). Although there was no preference for sea-bed terrain type by motherecalf pairs at their initial sighting, there was a preference for rugged terrain at their final resighting. A resource selection model indicated that the relative probability of a location being used by a motherecalf pair increased (as a function of water depth and rugged sea-bed terrain type) from initial to final sighting; a finding supported by subsequent tests of habitat preference versus availability. For 96 measured calves, calf length and water depth were positively correlated, even when ordinal day of measurement was controlled for statistically; a finding confirmed by a general linear model that simultaneously investigated the relationship between water depth, sea-bed terrain type, number of escorts, ordinal day and calf size. Thus, both calf age and size influence habitat choice by motherecalf pairs in their breeding grounds. The movement of mothers and their maturing calves into deeper waters where they favour rugged sea-bed terrain appears to be part of a suite of behavioural changes during the pre-migratory phase of residency in the breeding grounds