Establishment of mammalian cell lines containing multiple nonsense mutations and functional suppressor tRNA genes.

Journal ArticleWe describe the generation of mammalian cell lines carrying amber suppressor genes. Nonsense mutants in the herpes simplex virus thymidine kinase (HSV tk) gene, the Escherichia coli xanthine-guanine phosphoribosyl transferase (Eco-gpt) gene and the aminoglycoside 3' phosphotransferase gene of the Tn5 transposon (NPT-II) were isolated and characterized. Each gene was engineered with the appropriate control signals to allow expression in both E. coli and mammalian cells. Expression in E. coli made possible the use of well developed bacterial and phage genetic manipulations to isolate and characterize the nonsense mutants. Once characterized, the nonsense mutants were transferred into mammalian cells by microinjection and used, in turn, to select for amber suppressor genes. Xenopus laevis amber suppressor genes, prepared by site-specific mutagenesis of a normal X. laevis tRNA gene, were microinjected into the above cell lines and selected for the expression of one or more of the amber mutant gene products. The resulting cell lines, containing functional amber suppressor genes, are stable and exhibit normal growth rates

Why more boys than girls with ADHD receive treatment: A study of Dutch twins.

More boys than girls with attention deficit hyperactivity disorder (ADHD) receive treatment. One explanation for this bias may be that boys score higher on disruptive behavior scales than girls. Although this was supported by findings in clinical samples, recent studies in nonreferred samples showed that boys and girls with ADHD are similar with respect to their levels of disruptive behavior as reported by their mother. In this report, we investigate whether the difference in treatment rate is associated with higher teacher problem scores in boys with ADHD than in girls with ADHD. Data were obtained from mothers and teachers in a nonreferred sample of 283 boys and 291 girls with and without ADHD. Children were selected when they scored either low (controls) or high (probands) on attention problems. Mothers completed DSM-IV interviews, Child Behavior Checklists (CBCL) and the Conners Rating Scale (CRS). Teachers filled in the Teacher Report Form (TRF), and the CRS. Boys and girls with ADHD had similar levels of psychiatric illness and school impairment (such as being held back, special class placement and learning problems) by mother report. Mothers reported similar levels of aggression and attention problems in boys and girls with ADHD. In contrast, teachers consistently rated boys with ADHD as having higher scores on reports of attention problems and aggression than girls with ADHD. Gender differences vary across settings: boys and girls with ADHD are rated as behaving differently at school, but not at home. The higher level of teacher reported problem behavior at school may explain the high male-female ratio for ADHD in clinical settings. These findings have implications for the results of genetic studies that rely on referred samples, as these studies may give a distorted view of sex differences in the population

Structural changes and conductance thresholds in metal-free intrinsic SiOx resistive random access memory

We present an investigation of structural changes in silicon-rich silicon oxide metal-insulator-metal resistive RAM devices. The observed unipolar switching, which is intrinsic to the bulk oxide material and does not involve movement of metal ions, correlates with changes in the structure of the oxide. We use atomic force microscopy, conductive atomic force microscopy, x-ray photoelectron spectroscopy, and secondary ion mass spectroscopy to examine the structural changes occurring as a result of switching. We confirm that protrusions formed at the surface of samples during switching are bubbles, which are likely to be related to the outdiffusion of oxygen. This supports existing models for valence-change based resistive switching in oxides. In addition, we describe parallel linear and nonlinear conduction pathways and suggest that the conductance quantum, G0, is a natural boundary between the high and low resistance states of our devices

Prognosis of Hypothermic Patients Undergoing ECLS Rewarming-Do Alterations in Biochemical Parameters Matter?

While ECLS is a highly invasive procedure, the identification of patients with a potentially good prognosis is of high importance. The aim of this study was to analyse changes in the acid-base balance parameters and lactate kinetics during the early stages of ECLS rewarming to determine predictors of clinical outcome. This single-centre retrospective study was conducted at the Severe Hypothermia Treatment Centre at John Paul II Hospital in Krakow, Poland. Patients ≥18 years old who had a core temperature (Tc) < 30 °C and were rewarmed with ECLS between December 2013 and August 2018 were included. Acid-base balance parameters were measured at ECLS implantation, at Tc 30 °C, and at 2 and 4 h after Tc 30 °C. The alteration in blood lactate kinetics was calculated as the percent change in serum lactate concentration relative to the baseline. We included 50 patients, of which 36 (72%) were in cardiac arrest. The mean age was 56 ± 15 years old, and the mean Tc was 24.5 ± 12.6 °C. Twenty-one patients (42%) died. Lactate concentrations in the survivors group were significantly lower than in the non-survivors at all time points. In the survivors group, the mean lactate concentration decreased -2.42 ± 4.49 mmol/L from time of ECLS implantation until 4 h after reaching Tc 30 °C, while in the non-survivors' group (p = 0.024), it increased 1.44 ± 6.41 mmol/L. Our results indicate that high lactate concentration is associated with a poor prognosis for hypothermic patients undergoing ECLS rewarming. A decreased value of lactate kinetics at 4 h after reaching 30 °C is also associated with a poor prognosis

Genetic analyses of maternal and teacher ratings on attention problems in 7-year-old Dutch twins

The goal of the present study is to examine genetic and environmental influences on maternal and teacher ratings of Attention Problems (AP) in 7-year-old children. Teachers completed the Teacher Report Form (N=2259 pairs), and mothers the Child Behavior Checklist (N=2057 pairs). Higher correlations were found in twins rated by the same teacher than in twins rated by different teachers. This can be explained by rater bias or by a greater environmental sharing in twins, who are in the same classroom. We further found that 41% of the variation in maternal and teacher ratings is explained by a common factor. The heritability of this common factor is 78%. The heritabilities of the rater specific factors of mothers and teachers are 76% and 39%, respectively. Because Attention Problems that are persistent over situations may indicate more serious behavior problems than context dependent Attention Problems, we believe that gene finding strategies should focus on this common phenotype

HER2 testing in breast cancer: Opportunities and challenges

Human epidermal growth factor receptor 2 (HER2) is overexpressed in 15-25% of breast cancers, usually as a result of HER2 gene amplification. Positive HER2 status is considered to be an adverse prognostic factor. Recognition of the role of HER2 in breast cancer growth has led to the development of anti-HER2 directed therapy, with the humanized monoclonal antibody trastuzumab (Herceptin (R)) having been approved for the therapy of HER2-positive metastatic breast cancer. Clinical studies have further suggested that HER2 status can provide important information regarding success or failure of certain hormonal therapies or chemotherapies. As a result of these developments, there has been increasing demand to perform HER2 testing on current and archived breast cancer specimens. This article reviews the molecular background of HER2 function, activation and inhibition as well as current opinions concerning its role in chemosensitivity and interaction with estrogen receptor biology. The different tissue-based assays used to detect HER2 amplification and overexpression are discussed with respect to their advantages and disadvantages, when to test (at initial diagnosis or pre-treatment), where to test (locally or centralized) and the need for quality assurance to ensure accurate and valid testing results

The Latent Class Structure of ADHD is stable across informants

Previous studies have looked at the structure of attention-deficit/ hyperactivity disorder (ADHD) using latent class analysis (LCA) of Child Behavior Checklist (CBCL) or Diagnostic and Statistical Manual of Mental Disorders (DSM) symptom structure. These studies have identified distinct classes of children with inattentive, hyperactive, or combined subtypes and have used these classes to refine genetic analyses. The objective of the current report is to determine if the latent class structure of ADHD subtypes is consistent across informant using the Conners' Rating Scales (CRS). LCA was applied to CRS forms from mother, father, and teacher reports of 1837, 1329 and 1048 latency aged Dutch twins, respectively. The optimal solution for boys was a 5-class solution for mothers, a 3-class solution for fathers, and a 4-class solution for teachers. For girls, a 4-class solution for mothers and a 3-class for fathers and teachers was optimal. Children placed into a class by one informant had markedly increased odds ratio of being placed into the same or similar class by the other informants. Results from LCA using Dutch twins with the CRS show stability across informants suggesting that more stable phenotypes may be accessible for genotyping using a multi-informant approach

A study of genetic and environmental influences on maternal and paternal CBCL syndrome scores in a large sample of 3-year-old Dutch twins.

Background. There is increasing evidence that behavioral problems are common in very young children, yet little is known about the etiology of individual differences in these problems. It is unclear to what degree environmental and genetic factors influence the development of early child psychopathology. In this paper, we focus on the following issues. Firstly, to what degree do genetic and environmental factors influence variation in behavioral problems? Secondly, to what degree are these underlying etiological factors moderated by sex and informant? We investigate these issues by analyzing Child Behavior Checklist (CBCL) data on 9689 3-year-old twin pairs. Methods. Rater Bias and Psychometric Models were fitted to CBCL/2-3 data obtained from mothers and fathers to determine the genetic and environmental contributions to the five CBCL syndromes:aggressive, oppositional, overactive, withdrawn, and anxious/depressed behavior. Results. Parental ratings are influenced by aspects of the child's behavior that are experienced in the same way by both parents and by aspects of the child's behavior that are experienced uniquely by each parent. There is evidence for high genetic contributions to all CBCL syndromes. Shared and non-shared environmental influences play significant roles as well. One exception is overactive behavior, which is influenced by genetic and non-shared environmental influences only. Conclusions. Variation in behavior problems in the very young shows high heritability. Individual raters offer unique perspectives that can have an impact on estimates of problem behavior and genetic architecture. Therefore, multi-informant approaches in the assessment of the very young will be useful to clinicians and researchers alike

Influences on achieving motor milestones: A twin-singleton study

In order to determine if twinning impacted achievement of motor milestones the attainment of early motor milestones in twins was examined and compared to published data from singletons of the same age from the same culture and birth years. We examined the influence of twinning, sex, zygosity and birth cohort (1987-2001) on the motor development of twins aged 0 to 24 months. Data on the attainment of motor milestones (turn, sit, crawl, stand and walk) of twins were collected from maternal reports. All data were corrected for gestational age. Data from the twin sample were compared to normative data from singletons, which were available from Child Health Clinics (CHC). Analyses across twin data and the CHC singleton data revealed no differences between twins and singletons in achievement of motor milestones. Girls were able to sit without support slightly earlier than boys, otherwise there were no other sex differences. Birth-order analyses revealed minimal but significant differences in turning over from back to belly and for sitting without support between the first- and second-born. Dizygotic (DZ) twins were faster than monozygotic (MZ) twins in achieving the moment of sit, crawl, stand and walk. Twins born in earlier cohorts were faster in reaching the moment of crawl, sit and walk. It is concluded that there are no differences in time of reaching motor milestones between twins and singletons within the normal range. Sex has minimal to no effect on motor development in early childhood. DZ twins achieve motor milestones sooner than MZ twins. Attainment of gross motor milestones (crawl, stand and walk) is delayed in later birth cohorts

Resistive switching in silicon suboxide films

We report a study of resistive switching in a silicon-based memristor/resistive RAM (RRAM) device in which the active layer is silicon-rich silica. The resistive switching phenomenon is an intrinsic property of the silicon-rich oxide layer and does not depend on the diffusion of metallic ions to form conductive paths. In contrast to other work in the literature, switching occurs in ambient conditions, and is not limited to the surface of the active material. We propose a switching mechanism driven by competing field-driven formation and current-driven destruction of filamentary conductive pathways. We demonstrate that conduction is dominated by trap assisted tunneling through noncontinuous conduction paths consisting of silicon nanoinclusions in a highly nonstoichiometric suboxide phase. We hypothesize that such nanoinclusions nucleate preferentially at internal grain boundaries in nanostructured films. Switching exhibits the pinched hysteresis I/V loop characteristic of memristive systems, and on/off resistance ratios of 104:1 or higher can be easily achieved. Scanning tunneling microscopy suggests that switchable conductive pathways are 10 nm in diameter or smaller. Programming currents can be as low as 2 μA, and transition times are on the nanosecond scale