Action of bleomycin on structural mimics of intermediates in DNA double-strand cleavage

Bleomycin-induced cleavage was examined in several nicked, gapped, or intact duplex DNA substrates, including a structure designed to mimic a proposed singly nicked intermediate in double-strand cleavage. This nicked structure appeared to correctly target the second cleavage event in the complementary strand, resulting in a blunt-ended double-strand break, similar to that induced directly by bleomycin alone in an intact duplex of the same sequence. A one-base-gapped structure was markedly less efficient in correctly targeting bleomycin attack in the complementary strand. The results are consistent with a model of bleomycin-induced double-strand cleavage in which the nick formed by the initial bleomycin attack serves to target secondary attack to a specific position in the complementary strand, resulting in a double-strand break with a defined geometry.     

Tributary use and large-scale movements of grass carp in Lake Erie

Infrequent captures of invasive, non-native grass carp (Ctenopharyngodon idella) have occurred in Lake Erie over the last 30+ years, with recent evidence suggesting wild reproduction in the lake’s western basin (WB) is occurring. Information on grass carp movements in the Laurentian Great Lakes is lacking, but an improved understanding of large-scale movements and potential areas of aggregation will help inform control strategies and risk assessment if grass carp spread to other parts of Lake Erie and other Great Lakes. Twenty-three grass carp captured in Lake Erie’s WB were implanted with acoustic transmitters and released. Movements were monitored with acoustic receivers deployed throughout Lake Erie and elsewhere in the Great Lakes. Grass carp dispersed up to 236 km, with approximately 25% of fish dispersing greater than 100 km from their release location. Mean daily movements ranged from <0.01 to 2.49 km/day, with the highest daily averages occurring in the spring and summer. The Sandusky, Detroit, and Maumee Rivers, and Plum Creek were the most heavily used WB tributaries. Seventeen percent of grass carp moved into Lake Erie’s central or eastern basins, although all fish eventually returned to the WB. One fish emigrated from Lake Erie through the Huron-Erie Corridor and into Lake Huron. Based on our results, past assessments may have underestimated the potential for grass carp to spread in the Great Lakes. We recommend focusing grass carp control efforts on Sandusky River and Plum Creek given their high use by tagged fish, and secondarily on Maumee and Detroit Rivers.     

Spatial ecology of common carp (Cyprinus carpio) in Lake Winnipeg and its potential for management actions

Common carp feeding and spawning behaviours negatively impact the functioning of marsh ecosystems. In the Netley-Libau Marsh, situated on the southern end of Lake Winnipeg, water level regulations, nonpoint source nutrient pollution, and the non-native common carp are thought to be the main contributors to the degradation of the marsh habitat. Using acoustic telemetry, we analysed the movement rate, frequency and timing of suspected spawning migrations, spatial ecology, and aggregation of common carp in the Lake Winnipeg drainage over a three year time period. Common carp moved the farthest during the open water period when water temperature was >5 °C. Their annual migration into Netley-Libau Marsh was correlated to ordinal date. Common carp left the marsh in late spring/early summer, presumably feeding in Lake Winnipeg, before moving to overwintering sites situated in Traverse Bay and Lake Winnipeg, where they arrived typically by October and formed aggregations. These findings will inform habitat and fisheries managers in the effort to undertake evidence-based management actions. The predictability of the movements and the tendency for common carp to aggregate indicates that exclusion techniques and commercial fishing may represent viable management solutions.     

A probabilistic fire-protection siting model with joint vehicle reliability requirements

A probabilistic fire-protection siting model is described that places capacitated stations, engine companies, and truck companies in such a way that the population or calls covered by an engineand a truck with a joint reliability of at least is maximized. Probabilistic constraints are developed and numerical equivalents are found for the probability requirement for proximate server presence. The multiple co-location of servers at stations and the use of stations with a limited capacity are also investigated. Structures are utilized that preserve the integer properties when the model is solved by linear programming relaxation.An earlier version of this paper was presented at the 37th North American Meetings, Boston, November 1990.     

The Neurovascular Unit Dysfunction in Alzheimer’s Disease

Alzheimer’s disease (AD) is the most common neurodegenerative disease worldwide. Histopathologically, AD presents with two hallmarks: neurofibrillary tangles (NFTs), and aggregates of amyloid β peptide (Aβ) both in the brain parenchyma as neuritic plaques, and around blood vessels as cerebral amyloid angiopathy (CAA). According to the vascular hypothesis of AD, vascular risk factors can result in dysregulation of the neurovascular unit (NVU) and hypoxia. Hypoxia may reduce Aβ clearance from the brain and increase its production, leading to both parenchymal and vascular accumulation of Aβ. An increase in Aβ amplifies neuronal dysfunction, NFT formation, and accelerates neurodegeneration, resulting in dementia. In recent decades, therapeutic approaches have attempted to decrease the levels of abnormal Aβ or tau levels in the AD brain. However, several of these approaches have either been associated with an inappropriate immune response triggering inflammation, or have failed to improve cognition. Here, we review the pathogenesis and potential therapeutic targets associated with dysfunction of the NVU in AD.     

Insoluble Vascular Amyloid Deposits Trigger Disruption of the Neurovascular Unit in Alzheimer’s Disease Brains

Alzheimer’s disease (AD) is a neurodegenerative disease, characterized histopathologically by intra-neuronal tau-related lesions and by the accumulation of amyloid β-peptide (Aβ) in the brain parenchyma and around cerebral blood vessels. According to the vascular hypothesis of AD, an alteration in the neurovascular unit (NVU) could lead to Aβ vascular accumulation and promote neuronal dysfunction, accelerating neurodegeneration and dementia. To date, the effects of insoluble vascular Aβ deposits on the NVU and the blood–brain barrier (BBB) are unknown. In this study, we analyze different Aβ species and their association with the cells that make up the NVU. We evaluated post-mortem AD brain tissue. Multiple immunofluorescence assays were performed against different species of Aβ and the main elements that constitute the NVU. Our results showed that there are insoluble vascular deposits of both full-length and truncated Aβ species. Besides, insoluble aggregates are associated with a decrease in the phenotype of the cellular components that constitute the NVU and with BBB disruption. This approach could help identify new therapeutic targets against key molecules and receptors in the NVU that can prevent the accumulation of vascular fibrillar Aβ in AD.     

MicroRNAs in Metastasis and the Tumour Microenvironment

Metastasis is the process whereby cancer cells migrate from the primary tumour site to colonise the surrounding or distant tissue or organ. Metastasis is the primary cause of cancer-related mortality and approximately half of all cancer patients present at diagnosis with some form of metastasis. Consequently, there is a clear need to better understand metastasis in order to develop new tools to combat this process. MicroRNAs (miRNAs) regulate gene expression and play an important role in cancer development and progression including in the metastatic process. Particularly important are the roles that miRNAs play in the interaction between tumour cells and non-tumoral cells of the tumour microenvironment (TME), a process mediated largely by circulating miRNAs contained primarily in extracellular vesicles (EVs). In this review, we outline the accumulating evidence for the importance of miRNAs in the communication between tumour cells and the cells of the TME in the context of the pre-metastatic and metastatic niche.     

Molecular Mechanisms of the Deregulation of Muscle Contraction Induced by the R90P Mutation in Tpm3.12 and the Weakening of This Effect by BDM and W7

Point mutations in the genes encoding the skeletal muscle isoforms of tropomyosin can cause a range of muscle diseases. The amino acid substitution of Arg for Pro residue in the 90th position (R90P) in γ-tropomyosin (Tpm3.12) is associated with congenital fiber type disproportion and muscle weakness. The molecular mechanisms underlying muscle dysfunction in this disease remain unclear. Here, we observed that this mutation causes an abnormally high Ca²⁺-sensitivity of myofilaments in vitro and in muscle fibers. To determine the critical conformational changes that myosin, actin, and tropomyosin undergo during the ATPase cycle and the alterations in these changes caused by R90P replacement in Tpm3.12, we used polarized fluorimetry. It was shown that the R90P mutation inhibits the ability of tropomyosin to shift towards the outer domains of actin, which is accompanied by the almost complete depression of troponin’s ability to switch actin monomers off and to reduce the amount of the myosin heads weakly bound to F-actin at a low Ca²⁺. These changes in the behavior of tropomyosin and the troponin–tropomyosin complex, as well as in the balance of strongly and weakly bound myosin heads in the ATPase cycle may underlie the occurrence of both abnormally high Ca²⁺-sensitivity and muscle weakness. BDM, an inhibitor of myosin ATPase activity, and W7, a troponin C antagonist, restore the ability of tropomyosin for Ca²⁺-dependent movement and the ability of the troponin–tropomyosin complex to switch actin monomers off, demonstrating a weakening of the damaging effect of the R90P mutation on muscle contractility.     

Hydrogen embrittlement of high strength pipeline steels

A comparison was made between three API grade pipeline steels (X60, X80 and the X100 grade) from the point of view of their susceptibility to hydrogen embrittlement. The main aim was to determine whether the development of higher strength materials led to greater susceptibility to hydrogen embrittlement. This was achieved by straining at 2.8 × 10⁻⁵ s⁻¹ after cathodic charging. The results showed that there is a distinct susceptibility to loss of ductility after charging and this tends to increase with the strength level of the steel at a charging current density above 0.44 mA mm⁻². All three steels exhibited fine cracks parallel to the major rolling direction after charging and an increasing amount of brittleness on the fracture surface.