Simultaneous precise editing of multiple genes in human cells

Abstract. When double-strand breaks are introduced in a genome by CRISPR they are repaired either by non-homologous end joining (NHEJ), which often results i

Iron Deficiency Anemia, Population Health and Frailty in a Modern Portuguese Skeletal Sample

INTRODUCTION Portugal underwent significant political, demographic and epidemiological transitions during the 20th century resulting in migration to urban areas with subsequent overcrowding and issues with water sanitation. This study investigates population health during these transitions and interprets results within a framework of recent history and present-day public health information. We investigate skeletal evidence for anemia (cribra orbitalia and porotic hyperostosis) as indicators of stress and frailty-i.e., whether the lesions contribute to susceptibility for disease or increased risk of death. METHODS The presence and severity of skeletal lesions were compared against known sex and cause of death data to investigate potential heterogeneity in frailty and the relationship between lesions and risk of dying over time. Additionally, we tested for the presence of selective mortality in our data (i.e., whether or not the sample is biased for individuals with higher frailty). Our sample derives from a large, documented, modern Portuguese collection from Lisbon and is the first study of its kind using a documented collection. The collection represents primarily middle-class individuals. RESULTS AND CONCLUSIONS: Analyses indicated that porotic hyperostosis became more common and severe over time, while cribra orbitalia severity increased over time. Neither process was linked to cause of death. However, there was a significant relationship to sex; males exhibited a higher prevalence and severity of lesions and increased mortality. A Gompertz function showed decreased survivorship in early life but increased survivorship over age 60. Using comorbidities of anemia, we were unable to detect selective mortality-i.e., in our sample, lesions do not represent a sign of poor health or increased frailty and are not significantly linked with a decreased mean age-at-death. However, lesion prevalence and severity do reflect the socioeconomic processes in urban Lisbon during the 1800s and 1900s and the possibility of water-borne parasites as the contributing factor for iron deficiency anemia

Storage of spatiotemporal input sequences in dendrites of pyramidal neurons

Plastic changes in neurons are widely considered to underpin the formation and maintenance of memory. The mechanisms of induction and expression of plasticity are, therefore, crucial to our understanding of the capacity of information storage that neurons possess. Using two-photon glutamate uncaging and whole-cell electrophysiological recordings, I demonstrate that dendrites of neurons are capable of preferentially storing specific spatiotemporal sequences, and describe the physiological properties of this new form of plasticity. Such plastic changes are dependent on Ca2+ influx through NMDA receptors, which is consistent with previous reports regarding induction of potentiation. Using two-photon Ca2+ imaging, I demonstrate that spatiotemporal plasticity is a result of a distinct homogeneous spatial increase in Ca2+ influx of different spatiotemporal sequences. Using the NEURON simulation environment, I used my experimental findings to perform simulations of synaptic plasticity rules. I found that homogeneous increases in synaptic strength across the dendrite can result in the spatiotemporal plasticity that I empirically observed. Moreover, I employed a genetic optimization algorithm and parallelized simulations to show that such changes are within physiological parameters observed in cortical neurons. My PhD therefore describes a novel form of plasticity, and proposes that dendrites are capable of more extensive information storage than was previously assumed

Corrosion and Electrochemical Properties of Laser-Shock-Peening-Treated Stainless Steel AISI 304L in VVER Primary Water Environment

Laser Shock Peening (LSP) is a surface treatment technique for metallic materials. It induces plastic deformation at the surface of up to around 1 mm in depth. This process introduces residual stresses that lead to strain hardening, and potentially improvements in fatigue, stress corrosion cracking (SCC) and general corrosion behaviour in many, but not all, corrosive media. In this paper, two specimens made of AISI 304L stainless steel, one LSP-treated and one un-treated, were tested at 280 °C and 8 MPa in VVER (or PWR) primary circuit water chemistry using in situ Electrochemical Impedance Spectroscopy (EIS). This experiment serves to qualify the influence of LSP on the changes in corrosion behaviour in high-temperature, high-density water. The residual stress (RS) measurement of the surface showed a compression RS. Before LSP treatment, RS at the surface was 52.2 MPa in the rolling direction 0°RD and 10.42 MPa in the transverse rolling direction 90°RD. After the treatment, surface RS was −175.27 MPa and −183.51 MPa for Scan and TScan directions, respectively. The effect of compressive RS at the surface was studied and showed an increase in corrosion rate. The analysis of oxide layer by SEM revealed differences between LSP-treated and untreated AISI 304L specimens and their connection to corrosion rates

Knockdown of zif268 in the posterior dorsolateral striatum does not enduringly disrupt a response memory of a rewarded T-maze task

Under certain conditions pavlovian memories undergo reconsolidation, whereby the reactivated memory can be disrupted by manipulations such as knockdown of zif268. For instrumental memories, reconsolidation disruption is less well established. Our previous, preliminary data identified that there was an increase in Zif268 in the posterior dorsolateral striatum (pDLS) after expression of an instrumental habit-like ‘response’ memory, but not an instrumental goal-directed ‘place’ memory on a T-maze task. Here, the requirement for Zif268 in the reconsolidation of a response memory was tested by knockdown of Zif268, using antisense oligodeoxynucleotide infusion into the pDLS, at memory reactivation. Zif268 knockdown reduced response memory expression 72H, but not 7d later. Western blotting revealed a non-significant increase in Zif268 in the pDLS in rats using response memories, but there was no change in Zif268 expression in the hippocampus following retrieval of a place memory. Zif268 expression increased in the basolateral amygdala after memory reactivation whether a response or place strategy was used during reactivation. We propose that Zif268 expression in the basolateral amygdala may be linked to prediction error, generated by the absence of reward at reactivation. Taken together, these results suggest a complex role for Zif268 in the maintenance of instrumental memories.This work was conducted within the Behavioural and Clinical Neuroscience Institute (BCNI), a joint initiative funded by the Wellcome Trust and the UK Medical Research Council, in the Department of Psychology at the University of Cambridge. This work was funded by a UK Medical Research Council programme grant (no. G1002231) awarded to B.J.E. and A.L.M. G.H.V. was supported by a doctoral training grant from the BCNI. A.L.M. is the Ferreras-Willetts Fellow in Neuroscience at Downing College, Cambridge. E.N.C is a BBSRC Anniversary Future Leaders Fellow

TiO2 ALD Coating of Amorphous TiO2 Nanotube Layers: Inhibition of the Structural and Morphological Changes Due to Water Annealing

The present work presents a strategy to stabilize amorphous anodic self-organized TiO2 nanotube layers against morphological changes and crystallization upon extensive water soaking. The growth of needle-like nanoparticles was observed on the outer and inner walls of amorphous nanotube layers after extensive water soakings, in line with the literature on water annealing. In contrary, when TiO2 nanotube layers uniformly coated by thin TiO2 using atomic layer deposition (ALD) were soaked in water, the growth rates of needle-like nanoparticles were substantially reduced. We investigated the soaking effects of ALD TiO2 coatings with different thicknesses and deposition temperatures. Sufficiently thick TiO2 coatings (≈8.4 nm) deposited at different ALD process temperatures efficiently hamper the reactions between water and F− ions, maintain the amorphous state, and preserve the original tubular morphology. This work demonstrates the possibility of having robust amorphous 1D TiO2 nanotube layers that are very stable in water. This is very practical for diverse biomedical applications that are accompanied by extensive contact with an aqueous environment

Nanocrystalline diamond protects Zr cladding surface against oxygen and hydrogen uptake : Nuclear fuel durability enhancement

In this work, we demonstrate and describe an effective method of protecting zirconium fuel cladding against oxygen and hydrogen uptake at both accident and working temperatures in water-cooled nuclear reactor environments. Zr alloy samples were coated with nanocrystalline diamond (NCD) layers of different thicknesses, grown in a microwave plasma chemical vapor deposition apparatus. In addition to showing that such an NCD layer prevents the Zr alloy from directly interacting with water, we show that carbon released from the NCD film enters the underlying Zr material and changes its properties, such that uptake of oxygen and hydrogen is significantly decreased. After 100–170 days of exposure to hot water at 360 °C, the oxidation of the NCD-coated Zr plates was typically decreased by 40%. Protective NCD layers may prolong the lifetime of nuclear cladding and consequently enhance nuclear fuel burnup. NCD may also serve as a passive element for nuclear safety. NCD-coated ZIRLO claddings have been selected as a candidate for Accident Tolerant Fuel in commercially operated reactors in 2020

Field evaluation of controlled traffic farming in central Europe using commercially available machinery

The progressive increase in the size and weight of farm machinery causes concerns due to the increased risk of soil compaction that arises from non-organized vehicle traffic. Controlled traffic farming (CTF) offers an effective means to manage compaction by confining all load-bearing wheels to the least possible area of permanent traffic lanes. Although CTF is relatively well-established in Australia and in some countries in Northern Europe, its benefits and suitability for Central European conditions have not been demonstrated. A long-term experimental site was established in 2010 in Nitra, Slovakia, using a 6 m 'OutTrac-CTF' system with shallow non-inversion tillage practices. The 16 ha experimental field of loam soil is representative of land used for arable cropping in Central Europe. Four traffic intensities (non-trafficked, one traffic event per year with a single pass, multiple passes with permanent traffic lanes, and random traffic) were evaluated using two traffic systems: controlled (CTF) and non-controlled traffic farming (referred to as random traffic farming or RTF). This article reports the findings derived from the first four years of the project and focuses on the effects of traffic systems on yields observed in cereal crops (winter wheat, spring barley, and maize) grown at the site in a rotation cycle. Significant differences (p < 0.1) in yield are reported due to the heterogeneity of the field and the seasonal effect of weather. The results of this investigation suggest that CTF systems have potential to increase production sustainably in arable farming systems in Central Europe. Well-designed CTF systems using commercially available machinery allow for reductions in the area affected by traffic of up to 50% compared with random, non-organized traffic systems. Results also show that in years when soil moisture was not limiting, the yield penalty from a single (annual) machine pass was relatively small (~5%). However, in dry years, compaction caused by multiple machinery passes may lead to yield losses of up to 33%. When considering the ratio of non-trafficked to trafficked area within the different CTF systems evaluated in this study, yield improvements of up to 0.5 t ha-1 for cereals are possible when converting from RTF to CTF. Given the assumptions made in the analyses, such yield increases translate into increased revenues of up to 117 USD ha-1 (1 Euro= 1.1 USD). For Central European farming systems, the main benefit of CTF appears to be improved efficiency and enhanced agronomic stability, especially in dry seasons, where the significant yield penalty from machinery passes is likely