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The research presented in this essay is grounded in Social Domain Theory. Research provides substantial evidence that children's social development is characterized by the formation of distinctly different systems of thought, including those in the moral, social-conventional, and personal domains. A main focus here is on morality, defined as involving understandings of welfare, justice, and rights, which are applied across societal contexts. Social conventions are uniformities within social systems, serving to provide uniform expectations. The domains constitute different configurations of thinking and developmental changes occur within each domain. However, decisions in social situational contexts often involve coordination, which is a process of weighing and balancing different and sometimes conflicting considerations. Such social contexts can include conflicts between different moral goals or between moral and societal goals. Processes of coordination are examined in social psychological experiments, as well as developmental studies of topics like honesty, rights, and social inclusion. Coordination is also considered in people's perspectives on cultural practices of unfairness and inequality. Psychological research in patriarchal societies shows that females, who are subjected to inequalities evaluate those cultural practices as unfair. Anthropological research documents that females engage in acts of opposition and moral resistance regarding perceived unfair cultural practices.

Dissolved organic carbon (DOC) is important to microbial activity and nutrient cycling, and its concentration is sensitive to pH. Despite the importance of alkaline soils to agricultural production in southern Australia, few studies have documented the concentrations of soil organic carbon (C) and DOC or described the effects of soil properties and management practices on DOC in these soils. A survey of 33 paddocks from the Eyre Peninsula and mid-North regions of South Australia and north-western Victoria demonstrated significant variation in pH, soil organic C and DOC. Carbon stocks in the surface 30 cm were 40-55 t C/ha and were lowest in paddocks from Victoria. Soils from South Australia had higher DOC concentrations in the top 20 cm than soils from Victoria. Principal component analysis suggested variation in DOC was increased by high pH, electric conductivity and the concentration of exchangeable Na, and was reduced by the concentration of exchangeable Ca and clay content. Mid-infrared Fourier transform infrared spectroscopy identified regional differences in the composition of soil organic C, with high amounts of charcoal in Eyre Peninsula soils. Farm management practices had little effect on soil organic C but influenced DOC. Grain yield and DOC concentration were inversely related across and within regions which appeared to be related to the intensity of cropping having opposite influences on yield and DOC. Compared with international data, DOC concentrations were high relative to the amount of soil organic C and, in contrast to many previous studies, DOC in all regions increased with depth. Additional keywords: C : N ratio, carbon sequestration, farming system, sodic soil., Introduction A high proportion of the agricultural soils of southern Australia is alkaline (pH of 1 :5 soil water extract ([pH.sub.(1 : 5 water)] > 8) (dc Caritat et al. [...]

X-ray Absorption Fine Structure (XAFS) spectroscopy was used to identify directly the metal speciation and local bonding environment of Fe in sediments originated from Dobczyce reservoir (Poland); special attention was paid to analysis of samples collected from points situated at different distances from the land. The combination of traditional approach to data analysis as well as usage of multivariate techniques provided reliable information on the local Fe structure. The obtained average oxidation state for Fe species in all samples was found to be at about +3. The number, nature, and proportion of Fe species in sediments samples were calculated by a coupled Principal Component Analysis (PCA) and Least-Squares Fitting (LSF) procedure. Results indicate for a high similarity among analyzed data collected for various points from Dobczyce reservoir. Among different forms of amorphous oxyhydroxides and various iron oxides commonly present in lake sediments, hematite (Fe2O3) and goethite (α-FeOOH) were found to be the dominant crystalline structures in analyzed samples. Based on that finding it can be concluded that Fe is principally bound to oxygen in sediment samples, therefore its (re )mobilization to the water may be neglected. [ABSTRACT FROM AUTHOR]

Forensic science is a field that requires precise and reliable methods for the detection and analysis of evidence. One such method is Fourier Transform Infrared (FTIR) spectroscopy, which provides high sensitivity and selectivity in the detection of samples. In this study, the use of FTIR spectroscopy and statistical multivariate analysis to identify high explosive (HE) materials (C-4, TNT, and PETN) in the residues after high- and low-order explosions is demonstrated. Additionally, a detailed description of the data pre-treatment process and the use of various machine learning classification techniques to achieve successful identification is also provided. The best results were obtained with the hybrid LDA-PCA technique, which was implemented using the R environment, a code-driven open-source platform that promotes reproducibility and transparency.

ATR-FTIR (attenuated total reflection-Fourier-transform infrared) microscopy with imaging is widely used in the heritage field to characterise complex compositions of paint cross-sections. However, some limitations include the need for ATR crystal contact with the sample and the inability to resolve particle size below the IR diffraction limit. Recently, a novel O-PTIR (optical-photothermal infrared) spectroscopy technique claimed to open a new avenue for non-invasive, efficient, and reliable analysis at sub-micron resolution. O-PTIR produces transmission-like FTIR spectra for interpretation, without the need to touch the sample, which are highly favourable attributes for analysing heritage samples. This paper reports the comparison of O-PTIR and ATR-FTIR techniques applied to a cross-section embedding a thin paint fragment that delaminated from a late 19th to early 20th-century oil portrait. The hazy paint fragment consisted of zinc soaps (both crystalline and amorphous), gordaite (NaZn 4 Cl(OH) 6 SO 4 ·6H 2 O), and zinc lactate, that could not all be well-resolved with ATR-FTIR imaging. With O-PTIR analysis, the degradation compounds could be resolved at sub-micron resolution with an equivalent or better signal-to-noise ratio. This case study shows how the two techniques can be used to obtain comprehensive information at a broad level with ATR-FTIR and a detailed level with O-PTIR.

There is a considerable interest in developing new analytical tools to fight the illicit trafficking of heritage goods and particularly of easel paintings, whose high market values attract an ever-increasing volume of criminal activities. The objective is to combat the illicit traffic of smuggled or forged paintworks and to prevent the acquisition of fakes or looted artefacts in public collections. Authentication can be addressed using various investigation techniques, such as absolute dating, materials characterization, alteration phenomena, etc.; for paintings this remains a challenging task due to the complexity of the materials (paint layers, ground, varnish, canvas, etc.) and preferable use of non-destructive methods. This paper outlines results from concerted action on detecting forged works of art within the framework of a Coordinated Research Project of the International Atomic Energy Agency (IAEA) called Enhancing Nuclear Analytical Techniques to Meet the Needs of Forensic Sciences 1 . One of the main objectives is to foster the use of emerging Nuclear Analytical Techniques (NAT) using particle accelerators for authentication of paintings, with potential application to other forensics domains, by highlighting their ability to determine painting authenticity and to track restorations or anachronistic clues. The various materials comprising a test painting were investigated using an array of NAT. Binder, canvas and support were directly dated by 14 C using Accelerator Mass Spectrometry (14C-AMS); binder and pigments' molecular composition was determined using Secondary Ion Mass Spectrometry with MeV ions (MeV-SIMS); paint layer composition and stratigraphy were accurately determined using Ion Beam Analysis (IBA) and differential Particle-Induced X-ray Emission (PIXE); and pigment spatial distributions were mapped using full-field PIXE. High resolution Optical Photothermal Infrared Spectroscopy (O-PTIR) molecular imaging was also exploited. Obtained results are presented and discussed. It is shown that the combination of the above-mentioned techniques allowed reconstructing the history of the test painting., (Copyright © 2022. Published by Elsevier B.V.)

Malaria, caused by parasites of the species Plasmodium, is among the major life-threatening diseases to afflict humanity. The infectious cycle of Plasmodium is very complex involving distinct life stages and transitions characterized by cellular and molecular alterations. Therefore, novel single-cell technologies are warranted to extract details pertinent to Plasmodium-host cell interactions and underpinning biological transformations. Herein, we tested two emerging spectroscopic approaches: (a) Optical Photothermal Infrared spectroscopy and (b) Atomic Force Microscopy combined with infrared spectroscopy in contrast to (c) Fourier Transform InfraRed microspectroscopy, to investigate Plasmodium-infected erythrocytes. Chemical spatial distributions of selected bands and spectra captured using the three modalities for major macromolecules together with advantages and limitations of each method is presented here. These results indicate that O-PTIR and AFM-IR techniques can be explored for extracting sub-micron resolution molecular signatures within heterogeneous and dynamic samples such as Plasmodium-infected human RBCs., (© 2021. The Author(s).)

Antiferromagnetic insulators are a ubiquitous class of magnetic materials, holding the promise of low-dissipation spin-based computing devices that can display ultra-fast switching and are robust against stray fields. However, their imperviousness to magnetic fields also makes them difficult to control in a reversible and scalable manner. Here we demonstrate a novel proof-of-principle ionic approach to control the spin reorientation (Morin) transition reversibly in the common antiferromagnetic insulator α-Fe 2 O 3 (haematite) - now an emerging spintronic material that hosts topological antiferromagnetic spin-textures and long magnon-diffusion lengths. We use a low-temperature catalytic-spillover process involving the post-growth incorporation or removal of hydrogen from α-Fe 2 O 3 thin films. Hydrogenation drives pronounced changes in its magnetic anisotropy, Néel vector orientation and canted magnetism via electron injection and local distortions. We explain these effects with a detailed magnetic anisotropy model and first-principles calculations. Tailoring our work for future applications, we demonstrate reversible control of the room-temperature spin-state by doping/expelling hydrogen in Rh-substituted α-Fe 2 O 3 .

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-directed gene editing is approaching clinical implementation in cancer. Thus, it is imperative to define the molecular framework upon which safe and efficacious therapeutic strategies can be built. Two important reaction parameters include the biological time frame within which the CRISPR/Cas complex enters the nucleus and executes gene editing, and the method of discrimination that the CRISPR/Cas complex utilizes to target tumor cell, but not normal cell, genomes. We are developing CRISPR-directed gene editing for the treatment of non-small cell lung carcinoma focusing on disabling Nuclear Factor Erythroid 2-Related Factor-Like ( NRF2 ), a transcription factor that regulates chemoresistance and whose genetic disruption would enhance chemosensitivity. In this report, we define the time frame of cellular events that surround the initialization of CRISPR-directed gene editing as a function of the nuclear penetration and the execution of NRF2 gene disruption. We also identify a unique protospacer adjacent motif that facilitates site-specific cleavage of the NRF2 gene present only in tumor genomes. IMPLICATIONS: Our results begin to set a scientifically meritorious foundation for the exploitation of CRISPR-directed gene editing as an augmentative therapy for lung cancer and other solid tumors. VISUAL OVERVIEW: http://mcr.aacrjournals.org/content/molcanres/18/6/891/F1.large.jpg., (©2020 American Association for Cancer Research.)

Background: Agitation and aggression are commonly cited reasons for psychiatry consultation for individuals diagnosed with autism spectrum disorder (ASD). While risperidone and aripiprazole do not carry Health Canada approval for management of ASD-associated irritability, both are used for this indication but are not universally effective and carry substantial risk of adverse effects. This necessitates use of off-label medications to assist in management of behavioral dysregulation. Clonidine, an alpha-2 receptor agonist, is approved in Canada for treatment of hypertension. The evidence base also supports its use for attention deficit/hyperactivity disorder (ADHD) and for tics in Tourette's disorder. This review focuses on examining the literature regarding clonidine as a treatment of challenging behaviours in the ASD population., Method: Systematic search of MEDLINE, EMBASE, and PsycINFO databases resulted in 540 unique records. Ten publications were relevant to this review., Results: Two cross-over studies, one open-label case series, and seven case reports were identified. One of two controlled studies suggested benefit from clonidine versus placebo. Caregivers typically noted improvement in behaviour with clonidine versus baseline. Clonidine was generally well-tolerated. Sedation was the most consistently reported adverse effect. Despite being an anti-hypertensive medication, few discontinued clonidine due to hypotension or bradycardia., Conclusion: Clonidine has a limited evidence base for use in the management of behavioural problems in patients with ASD. Most evidence originates from case reports. Given the paucity of pharmacological options for addressing challenging behaviours in ASD patients, a clonidine trial may be an appropriate and cost-effective pharmaceutical option for this population., (Copyright © 2020 Canadian Academy of Child and Adolescent Psychiatry.)

Recent studies point to the evolution of drug resistance in lung cancer as being centered, at least in part, on the upregulation of various genes involved in controlling efflux or drug inactivation. Among the most important of these genes is Nuclear Factor Erythroid 2-Related Factor ( NRF2 ), considered the master regulator of 100-200 target genes involved in cellular responses to oxidative and/or electrophilic stress. With increased focus on the development of combinatorial approaches for cancer treatment, we utilized CRISPR/Cas9 to disable the NRF2 gene in lung cancer cells by disrupting the NRF2 nuclear export signal (NES) domain; phenotypically, the protein is largely blocked from transiting into the nucleus after translation. In tissue culture, cells with this gene knockout were found to have a reduced proliferation phenotype and are more sensitive to chemotherapeutic agents, such as cisplatin and carboplatin. These observations were confirmed in xenograft mouse models wherein the homozygous knockout cells proliferate at a slower rate than the wild-type cells, even in the absence of drug treatment. Tumor growth was arrested for a period of 16 days, with a dramatic decrease in tumor volume being observed in samples receiving the combined action of CRISPR-directed gene editing and chemotherapy.

Background: Lone parents in high-income countries have high rates of poverty (including in-work poverty) and poor health. Employment requirements for these parents are increasingly common. 'Welfare-to-work' (WtW) interventions involving financial sanctions and incentives, training, childcare subsidies and lifetime limits on benefit receipt have been used to support or mandate employment among lone parents. These and other interventions that affect employment and income may also affect people's health, and it is important to understand the available evidence on these effects in lone parents., Objectives: To assess the effects of WtW interventions on mental and physical health in lone parents and their children living in high-income countries. The secondary objective is to assess the effects of welfare-to-work interventions on employment and income., Search Methods: We searched the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE Ovid, Embase Ovid, PsycINFO EBSCO, ERIC EBSCO, SocINDEX EBSCO, CINAHL EBSCO, Econlit EBSCO, Web of Science ISI, Applied Social Sciences Index and Abstracts (ASSIA) via Proquest, International Bibliography of the Social Sciences (IBSS) via ProQuest, Social Services Abstracts via Proquest, Sociological Abstracts via Proquest, Campbell Library, NHS Economic Evaluation Database (NHS EED) (CRD York), Turning Research into Practice (TRIP), OpenGrey and Planex. We also searched bibliographies of included publications and relevant reviews, in addition to many relevant websites. We identified many included publications by handsearching. We performed the searches in 2011, 2013 and April 2016., Selection Criteria: Randomised controlled trials (RCTs) of mandatory or voluntary WtW interventions for lone parents in high-income countries, reporting impacts on parental mental health, parental physical health, child mental health or child physical health., Data Collection and Analysis: One review author extracted data using a standardised extraction form, and another checked them. Two authors independently assessed risk of bias and the quality of the evidence. We contacted study authors to obtain measures of variance and conducted meta-analyses where possible. We synthesised data at three time points: 18 to 24 months (T1), 25 to 48 months (T2) and 49 to 72 months (T3)., Main Results: Twelve studies involving 27,482 participants met the inclusion criteria. Interventions were either mandatory or voluntary and included up to 10 discrete components in varying combinations. All but one study took place in North America. Although we searched for parental health outcomes, the vast majority of the sample in all included studies were female. Therefore, we describe adult health outcomes as 'maternal' throughout the results section. We downgraded the quality of all evidence at least one level because outcome assessors were not blinded. Follow-up ranged from 18 months to six years. The effects of welfare-to-work interventions on health were generally positive but of a magnitude unlikely to have any tangible effects.At T1 there was moderate-quality evidence of a very small negative impact on maternal mental health (standardised mean difference (SMD) 0.07, 95% Confidence Interval (CI) 0.00 to 0.14; N = 3352; studies = 2)); at T2, moderate-quality evidence of no effect (SMD 0.00, 95% CI 0.05 to 0.05; N = 7091; studies = 3); and at T3, low-quality evidence of a very small positive effect (SMD -0.07, 95% CI -0.15 to 0.00; N = 8873; studies = 4). There was evidence of very small positive effects on maternal physical health at T1 (risk ratio (RR) 0.85, 95% CI 0.54 to 1.36; N = 311; 1 study, low quality) and T2 (RR 1.06, 95% CI 0.95 to 1.18; N = 2551; 2 studies, moderate quality), and of a very small negative effect at T3 (RR 0.97, 95% CI 0.91 to 1.04; N = 1854; 1 study, low quality).At T1, there was moderate-quality evidence of a very small negative impact on child mental health (SMD 0.01, 95% CI -0.06 to 0.09; N = 2762; studies = 1); at T2, of a very small positive effect (SMD -0.04, 95% CI -0.08 to 0.01; N = 7560; studies = 5), and at T3, there was low-quality evidence of a very small positive effect (SMD -0.05, 95% CI -0.16 to 0.05; N = 3643; studies = 3). Moderate-quality evidence for effects on child physical health showed a very small negative effect at T1 (SMD -0.05, 95% CI -0.12 to 0.03; N = 2762; studies = 1), a very small positive effect at T2 (SMD 0.07, 95% CI 0.01 to 0.12; N = 7195; studies = 3), and a very small positive effect at T3 (SMD 0.01, 95% CI -0.04 to 0.06; N = 8083; studies = 5). There was some evidence of larger negative effects on health, but this was of low or very low quality.There were small positive effects on employment and income at 18 to 48 months (moderate-quality evidence), but these were largely absent at 49 to 72 months (very low to moderate-quality evidence), often due to control group members moving into work independently. Since the majority of the studies were conducted in North America before the year 2000, generalisabilty may be limited. However, all study sites were similar in that they were high-income countries with developed social welfare systems., Authors' Conclusions: The effects of WtW on health are largely of a magnitude that is unlikely to have tangible impacts. Since income and employment are hypothesised to mediate effects on health, it is possible that these negligible health impacts result from the small effects on economic outcomes. Even where employment and income were higher for the lone parents in WtW, poverty was still high for the majority of the lone parents in many of the studies. Perhaps because of this, depression also remained very high for lone parents whether they were in WtW or not. There is a lack of robust evidence on the health effects of WtW for lone parents outside North America.

Objective: The goal of this contribution is to present and familiarize the medical community with the method for the assessment of trace and essentials elements in prostate tissue sections., Materials and Methods: X-ray fluorescence based technique(namely Synchrotron Induced X-ray Emission (SRIXE)) is described in terms of methodology, sample preparation and the evaluation of the recorded results (spectral data sets). Materials for the samples were collected from the patients underwent radical prostatectomy due to Adenocarcinoma prostatae. Specimens were freeze-dried, cut by microtome (to the thickness of 15 μm), one slice was placed on Mylar foil (for SRIXE measurements) and adjacent one on microscopic glass (for histopathological assessment)., Results: Results presented here show the usability of SRIXE method for the evaluation of concentration of trace and essential elements in prostate tissue sections with the spatial resolution better than 15 microns., Discussion: Histopathological analysis of samples, which is only focused on morphological features, is unable to reveal information about changes in biochemical signature of tissues affected by the illness. SRIXE is a powerful and promising technique to analyse even very low concentrations oat the cellular level without any labelling or separating procedures. Obtained results may be correlated with classic histopathological assessment allowing for drawing conclusions on the changes in certain elements concentrations with the progression of disease. Moreover, mentioned in this work analysis, can be performed for any type of biological tissues.

Background: Lone parents in high-income countries have high rates of poverty (including in-work poverty) and poor health. Employment requirements for these parents are increasingly common. 'Welfare-to-work' (WtW) interventions involving financial sanctions and incentives, training, childcare subsidies and lifetime limits on benefit receipt have been used to support or mandate employment among lone parents. These and other interventions that affect employment and income may also affect people's health, and it is important to understand the available evidence on these effects in lone parents., Objectives: To assess the effects of WtW interventions on mental and physical health in lone parents and their children living in high-income countries. The secondary objective is to assess the effects of welfare-to-work interventions on employment and income., Search Methods: We searched the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE Ovid, Embase Ovid, PsycINFO EBSCO, ERIC EBSCO, SocINDEX EBSCO, CINAHL EBSCO, Econlit EBSCO, Web of Science ISI, Applied Social Sciences Index and Abstracts (ASSIA) via Proquest, International Bibliography of the Social Sciences (IBSS) via ProQuest, Social Services Abstracts via Proquest, Sociological Abstracts via Proquest, Campbell Library, NHS Economic Evaluation Database (NHS EED) (CRD York), Turning Research into Practice (TRIP), OpenGrey and Planex. We also searched bibliographies of included publications and relevant reviews, in addition to many relevant websites. We identified many included publications by handsearching. We performed the searches in 2011, 2013 and April 2016., Selection Criteria: Randomised controlled trials (RCTs) of mandatory or voluntary WtW interventions for lone parents in high-income countries, reporting impacts on parental mental health, parental physical health, child mental health or child physical health., Data Collection and Analysis: One review author extracted data using a standardised extraction form, and another checked them. Two authors independently assessed risk of bias and the quality of the evidence. We contacted study authors to obtain measures of variance and conducted meta-analyses where possible. We synthesised data at three time points: 18 to 24 months (T1), 25 to 48 months (T2) and 49 to 72 months (T3)., Main Results: Twelve studies involving 27,482 participants met the inclusion criteria. Interventions were either mandatory or voluntary and included up to 10 discrete components in varying combinations. All but one study took place in North America. Although we searched for parental health outcomes, the vast majority of the sample in all included studies were female. Therefore, we describe adult health outcomes as 'maternal' throughout the results section. We downgraded the quality of all evidence at least one level because outcome assessors were not blinded. Follow-up ranged from 18 months to six years. The effects of welfare-to-work interventions on health were generally positive but of a magnitude unlikely to have any tangible effects.At T1 there was moderate-quality evidence of a very small negative impact on maternal mental health (standardised mean difference (SMD) 0.07, 95% Confidence Interval (CI) 0.00 to 0.14; N = 3352; studies = 2)); at T2, moderate-quality evidence of no effect (SMD 0.00, 95% CI 0.05 to 0.05; N = 7091; studies = 3); and at T3, low-quality evidence of a very small positive effect (SMD -0.07, 95% CI -0.15 to 0.00; N = 8873; studies = 4). There was evidence of very small positive effects on maternal physical health at T1 (risk ratio (RR) 0.85, 95% CI 0.54 to 1.36; N = 311; 1 study, low quality) and T2 (RR 1.06, 95% CI 0.95 to 1.18; N = 2551; 2 studies, moderate quality), and of a very small negative effect at T3 (RR 0.97, 95% CI 0.91 to 1.04; N = 1854; 1 study, low quality).At T1, there was moderate-quality evidence of a very small negative impact on child mental health (SMD 0.01, 95% CI -0.06 to 0.09; N = 2762; studies = 1); at T2, of a very small positive effect (SMD -0.04, 95% CI -0.08 to 0.01; N = 7560; studies = 5), and at T3, there was low-quality evidence of a very small positive effect (SMD -0.05, 95% CI -0.16 to 0.05; N = 3643; studies = 3). Moderate-quality evidence for effects on child physical health showed a very small negative effect at T1 (SMD -0.05, 95% CI -0.12 to 0.03; N = 2762; studies = 1), a very small positive effect at T2 (SMD 0.07, 95% CI 0.01 to 0.12; N = 7195; studies = 3), and a very small positive effect at T3 (SMD 0.01, 95% CI -0.04 to 0.06; N = 8083; studies = 5). There was some evidence of larger negative effects on health, but this was of low or very low quality.There were small positive effects on employment and income at 18 to 48 months (moderate-quality evidence), but these were largely absent at 49 to 72 months (very low to moderate-quality evidence), often due to control group members moving into work independently. Since the majority of the studies were conducted in North America before the year 2000, generalisabilty may be limited. However, all study sites were similar in that they were high-income countries with developed social welfare systems., Authors' Conclusions: The effects of WtW on health are largely of a magnitude that is unlikely to have tangible impacts. Since income and employment are hypothesised to mediate effects on health, it is possible that these negligible health impacts result from the small effects on economic outcomes. Even where employment and income were higher for the lone parents in WtW, poverty was still high for the majority of the lone parents in many of the studies. Perhaps because of this, depression also remained very high for lone parents whether they were in WtW or not. There is a lack of robust evidence on the health effects of WtW for lone parents outside North America.

CRISPR/Cas9 and single-stranded DNA oligonucleotides (ssODNs) have been used to direct the repair of a single base mutation in human genes. Here, we examine a method designed to increase the precision of RNA guided genome editing in human cells by utilizing a CRISPR/Cas9 ribonucleoprotein (RNP) complex to initiate DNA cleavage. The RNP is assembled in vitro and induces a double stranded break at a specific site surrounding the mutant base designated for correction by the ssODN. We use an integrated mutant eGFP gene, bearing a single base change rendering the expressed protein nonfunctional, as a single copy target in HCT 116 cells. We observe significant gene correction activity of the mutant base, promoted by the RNP and single-stranded DNA oligonucleotide with validation through genotypic and phenotypic readout. We demonstrate that all individual components must be present to obtain successful gene editing. Importantly, we examine the genotype of individually sorted corrected and uncorrected clonally expanded cell populations for the mutagenic footprint left by the action of these gene editing tools. While the DNA sequence of the corrected population is exact with no adjacent sequence modification, the uncorrected population exhibits heterogeneous mutagenicity with a wide variety of deletions and insertions surrounding the target site. We designate this type of DNA aberration as on-site mutagenicity. Analyses of two clonal populations bearing specific DNA insertions surrounding the target site, indicate that point mutation repair has occurred at the level of the gene. The phenotype, however, is not rescued because a section of the single-stranded oligonucleotide has been inserted altering the reading frame and generating truncated proteins. These data illustrate the importance of analysing mutagenicity in uncorrected cells. Our results also form the basis of a simple model for point mutation repair directed by a short single-stranded DNA oligonucleotides and CRISPR/Cas9 ribonucleoprotein complex., Competing Interests: The authors have declared that no competing interests exist.

In this paper, we present the first experimental demonstration of double- and triple-band negative refraction index meta-foils in the terahertz (THz) region. Multi-band meta-foils constructed by multi-cell S-string resonators in a single structure exhibit simultaneously negative permittivity and negative permeability responses at multiple frequencies. The phenomena are confirmed by numerical simulations and Fourier transform infrared spectroscopy measurements. The flexible, freestanding multi-band meta-foils provide a promising candidate for the development of multi-frequency THz materials and devices.

It has long been suggested that in skeletal muscle, the ATP-sensitive K(+) channel (K(ATP)) channel is important in protecting energy levels and that abolishing its activity causes fiber damage and severely impairs function. The responses to a lack of K(ATP) channel activity vary between muscles and fibers, with the severity of the impairment being the highest in the most glycolytic muscle fibers. Furthermore, glycolytic muscle fibers are also expected to face metabolic stress more often than oxidative ones. The objective of this study was to determine whether the t-tubular K(ATP) channel content differs between muscles and fiber types. K(ATP) channel content was estimated using a semiquantitative immunofluorescence approach by staining cross sections from soleus, extensor digitorum longus (EDL), and flexor digitorum brevis (FDB) muscles with anti-Kir6.2 antibody. Fiber types were determined using serial cross sections stained with specific antimyosin I, IIA, IIB, and IIX antibodies. Changes in Kir6.2 content were compared with changes in CaV1.1 content, as this Ca(2+) channel is responsible for triggering Ca(2+) release from sarcoplasmic reticulum. The Kir6.2 content was the lowest in the oxidative soleus and the highest in the glycolytic EDL and FDB. At the individual fiber level, the Kir6.2 content within a muscle was in the order of type IIB > IIX > IIA ≥ I. Interestingly, the Kir6.2 content for a given fiber type was significantly different between soleus, EDL, and FDB, and highest in FDB. Correlations of relative fluorescence intensities from the Kir6.2 and CaV1.1 antibodies were significant for all three muscles. However, the variability in content between the three muscles or individual fibers was much greater for Kir6.2 than for CaV1.1. It is suggested that the t-tubular K(ATP) channel content increases as the glycolytic capacity increases and as the oxidative capacity decreases and that the expression of K(ATP) channels may be linked to how often muscles/fibers face metabolic stress.

Activation of the K(ATP) channels results in faster fatigue rates as the channels depress action potential amplitude, whereas abolishing the channel activity has no effect in whole extensor digitorum longus (EDL) and soleus muscles. In this study, we examined the effects of abolished K(ATP) channel activity during fatigue at 37 degrees C on free intracellular Ca(2+) (Ca(2+)(i)) and tetanic force using single muscle fibres and small muscle bundles from the flexor digitorum brevis (FDB). K(ATP) channel deficient muscle fibres were obtained (i) pharmacologically by exposing wild-type fibres to glibenclamide, and (ii) genetically using null mice for the Kir6.2 gene (Kir6.2(-/-) mice). Fatigue was elicited using 200 ms tetanic contractions every second for 3 min. This study demonstrated for the first time that abolishing K(ATP) channel activity at 37 degrees C resulted in faster fatigue rates, where decreases in peak Ca(2+)(i) and tetanic force were faster in K(ATP) channel deficient fibres than in control wild-type fibres. Furthermore, several contractile dysfunctions were also observed in K(ATP) channel deficient muscle fibre. They included partially or completely supercontracted single muscle fibres, greater increases in unstimulated Ca(2+)(i) and unstimulated force, and lower force recovery. We propose that the observed faster rate of fatigue in K(ATP) channel deficient fibres is because the decreases in peak Ca(2+)(i) and force caused by contractile dysfunctions prevail over the expected slower decreases when the channels do not depress action potential amplitude.