Toxicity testing of human assisted reproduction devices using the mouse embryo assay.

Systems to assess the toxicity of materials used in human assisted reproduction currently lack efficiency and/or sufficient discriminatory power. The development of 1-cell CBA/B6 F1 hybrid mouse embryos to blastocysts, expressed as blastocyst rate (BR), is used to measure toxicity. The embryos were divided into control and test groups, and were exposed to either control medium or to a potentially toxic test medium. Inferences on toxicity were based on differences in BR between the two groups. The mouse embryo assay followed a stratified (mouse), randomized (embryo), and balanced (equal number of embryos per group and per mouse) design. The number of embryos needed was calculated using power analysis. The basal BR of the hybrid strain was determined in a historical population. Sixty-nine mouse embryos per group were required to detect toxic materials with sufficient sensitivity and to account for the considerable inter-mouse variation in blastocyst development. Fifty-two samples, divided over batches of seven different products were tested before use in the study IVF centre and five of these were found to be toxic. This test system, presented as the Nijmegen mouse embryo assay (NMEA), can be used to detect embryo-toxic materials in daily IVF practice, and this report may provide a starting point for standardization.

Seeing baby: women's experience of prenatal ultrasound examination and unexpected fetal diagnosis.

OBJECTIVE: Although prenatal ultrasound (US) is a common clinical undertaking today, little information is available about women's experience of the procedure from the perspective of women themselves. The objective of this study was to explore women's experience of undergoing a routine prenatal US examination associated with an unexpected fetal diagnosis. STUDY DESIGN: Qualitative methods were used to explore the prenatal US experience of 13 women. Five women were given unexpected news of multiple pregnancy and eight women were given unexpected news of congenital fetal abnormality. One in-depth audio-taped interview was conducted with each woman. Content analysis of interview data identified themes common to women's experience of US. RESULTS: Identified themes of women's experience of routine prenatal US examination associated with an unexpected fetal diagnosis are: experiencing the setting, sensing information, feeling connected/disconnected, the power of the image, and communication rules. CONCLUSIONS: Women's experience of prenatal US examination is influenced by physical and environmental factors and by the behaviors of the US examiner. Behaviors of the examiner contribute to a woman's labeling of the US experience as positive or negative. Women identify being objectified by the examination and experience poor communication patterns after a fetal US diagnosis. Women's description of the US screen image as a baby suggests it is a powerful influence on subsequent clinical and ethical decision-making about the pregnancy.

Building safer systems by ecological design: using restoration science to develop a medication safety intervention.

BACKGROUND: Experts call for stronger safety cultures and transparent reporting practices to increase medication safety in today's strained healthcare environments. The field of ecological restoration is concerned with the effective, efficient, and sustainable repair and recovery of ecosystems that have been degraded, damaged, or destroyed. A study was undertaken to determine whether the lessons of restoration science can be adapted to the study of medication safety issues. METHODS: Working with 26 practitioners, the principles of good restoration were used to design and pilot an innovative multifaceted medication safety intervention. The intervention included focus groups with practitioners, the construction and administration of a research based medication safety inventory, repeat digital photography of environmental safety issues, and targeted environmental modifications. RESULTS: Participants were most concerned about staff education and the physical environment for medication administration. Ward staff used the research to build a healthy reporting culture, introduce regular discussions of near misses, develop education strategies, redesign delivery and storage processes, and renovate the environment. CONCLUSIONS: Members of a busy hospital ward successfully adapted methods of restoration science to study, redesign, and strengthen medication safety practices and ward safety culture within existing resources. Further research will be conducted to test the merits of restoration science for health care.

Sample size calculations for a split-cluster, beta-binomial design in the assessment of toxicity.

Mouse embryo assays are recommended to test materials used for in vitro fertilization for toxicity. In such assays, a number of embryos is divided in a control group, which is exposed to a neutral medium, and a test group, which is exposed to a potentially toxic medium. Inferences on toxicity are based on observed differences in successful embryo development between the two groups. However, mouse embryo assays tend to lack power due to small group sizes. This paper focuses on the sample size calculations for one such assay, the Nijmegen mouse embryo assay (NMEA), in order to obtain an efficient and statistically validated design. The NMEA follows a stratified (mouse), randomized (embryo), balanced design (also known as a split-cluster design). We adopted a beta-binomial approach and obtained a closed sample size formula based on an estimator for the within-cluster variance. Our approach assumes that the average success rate of the mice and the variance thereof, which are breed characteristics that can be easily estimated from historical data, are known. To evaluate the performance of the sample size formula, a simulation study was undertaken which suggested that the predicted sample size was quite accurate. We confirmed that incorporating the a priori knowledge and exploiting the intra-cluster correlations enable a smaller sample size. Also, we explored some departures from the beta-binomial assumption. First, departures from the compound beta-binomial distribution to an arbitrary compound binomial distribution lead to the same formulas, as long as some general assumptions hold. Second, our sample size formula compares to the one derived from a linear mixed model for continuous outcomes in case the compound (beta-)binomial estimator is used for the within-cluster variance.

Hermeneutic-phenomenology: providing living knowledge for nursing practice.

The phenomenological approach has gained popularity among nurse researchers as an alternative investigative method to those used in the natural sciences. As more nurse scholars and nurse researchers utilize phenomenology as a research approach, it becomes critical to examine the implications this may have for nursing knowledge development and for the utilization of that knowledge in practice. In this paper, an examination of the results of phenomenological inquiry is presented and compared with the types of knowledge considered important for nursing by Carper and White. It is clear that phenomenology contributes to empirical, moral, aesthetic, personal, and socio-political knowledge development. Its contribution is not in developing predictive and prescriptive theory, but in revealing the nature of human experience. Although interpretive inquiry, such as hermeneutic phenomenology, does not prescribe action for use in clinical practice, it does influence a thoughtful reflective attentive practice by its revealing of the meanings of human experience.

The perinatal death of a twin. Karla's story of attaching and detaching.

This article examines the attaching and detaching experience of a mother encountering the perinatal death of a twin. Her experience is related to the relevant theoretical and research literature pertaining to prebirth and postbirth maternal-infant attachment and detachment (grieving). Literature for both single infants and twins is considered. The experience of this mother suggests that elements of postbirth attachment may have been accelerated into the prebirth period. In addition, her postbirth attaching and detaching experience suggests that an attachment and detachment to the twins as a unit preceded detachment from the twin who died. The health care provider's role in promoting maternal well-being, and indirectly the well-being of the surviving infant, is described.

Accommodating a twin pregnancy: maternal processes.

The object of this study was to develop a theory of the relationship between a mother and her unborn twins using a qualitative grounded theory methodology. Ten women participated in interviews during the last trimester of a twin pregnancy and again during the early postpartum period. Data provided by these women was analysed using the constant comparative method. Two additional informants were sampled in order to formulate and verify the tentative theory. For women in this study, the findings indicate that mothers from a relationship with their unborn twins within the wider context of accommodating the twin pregnancy in their lives. A woman accomplishes this process by immediately seeking information about twin pregnancy, by making room, in all senses, for two babies instead of one in her life, and by appraising her own pregnancy risk. A mother engages in self-protective behaviors and in behaviors designed to protect the health of her unborn twins as a method of coping with the perceived risk of the twin pregnancy and with the twin pregnancy itself. As the twin pregnancy progresses, women move toward accepting the idea of being pregnant with twins. For women pregnant with twins, the birth of two infants confirms the reality of the pregnancy.

Increase of sodium current after pyrethroid insecticides in mouse neuroblastoma cells.

The effects of 4 different pyrethroid insecticides on sodium channel gating in internally perfused, cultured mouse neuroblastoma cells (N1E-115) were studied using the suction pipette, voltage clamp technique. Pyrethroids increased the amplitude of the sodium current, sometimes by more than 200%. Activation of the sodium current occurred at more hyperpolarized potentials than under control conditions. The declining phase of the sodium current during depolarization was markedly slowed down and after repolarization of the membrane a large, slowly decaying sodium tail current developed. Pyrethroids did not affect the sodium current reversal potential, steady-state sodium inactivation or recovery from sodium channel inactivation. The amplitude of the pyrethroid-induced slow tail current was always proportional to the sodium current at the end of the preceding depolarizing pulse. The rate of decay of the slow tail current strongly depended on pyrethroid structure and increased in the order deltamethrin, cyphenothrin, fenfluthrin and phenothrin. The rate of decay further depended on membrane potential and temperature. Below -85 m V the instantaneous current-voltage relationship of the slow tail current showed a negative slope conductance. The tail current decayed more slowly at low temperatures. Arrhenius plots indicated that the relaxation of open sodium channels to a closed state involved a higher energy barrier for pyrethroid-affected than for normal channels. The energy barrier was higher after deltamethrin than after the non-cyano pyrethroid fenfluthrin. It is concluded that in mammalian neuronal membrane pyrethroids selectively reduce the rate of closing of sodium channels both during depolarization and after repolarization of the nerve membrane.

Temperature- and structure-dependent interaction of pyrethroids with the sodium channels in frog node of Ranvier.

(1) The interaction of a series of pyrethroid insecticides with the Na+ channels in myelinated nerve fibres of the clawed frog, Xenopus laevis, was investigated using the voltage clamp technique. (2) Out of 11 pyrethroids 9 insecticidally active compounds induce a slowly decaying Na+ tail current on termination of a step depolarization, whereas the Na+ current during depolarization was hardly affected. These tail currents are most readily explained by a selective reduction of the rate of closing of the activation gate in a fraction of the Na+ channels that have opened during depolarization. (3) The rate of decay of the Na+ tail current varies considerably with pyrethroid structure. After alpha-cyano pyrethroids the decay is at least one order of magnitude slower than after non-cyano pyrethroids. The decay always follows a single-exponential time course and is reversibly slowed when the temperature is lowered from 25 to 0 degrees C. Arrhenius plots in this temperature range are linear. (4) These results indicate that the relaxation of the activation gate in pyrethroid-affected Na+ channels is governed by an apparent first order, unimolecular process and that the rate of relaxation is limited by a single energy barrier. Application of transition state theory shows that after alpha-cyano pyrethroids this energy barrier is 9.6 kJ/mol higher than after non-cyano pyrethroids. (5) Differences in rate of decay of the Na+ tail current account for the reported differences in repetitive nerve activity induced by various pyrethroids. In addition, the effect of temperature on the rate of decay explains the increase in repetitive activity with cooling.

Frequency response of the lateral-line organ of Xenopus laevis.

The stimulus response relation of the epidermal lateral-line organ of Xenopus laevis was studied by recording activity of single afferent nerve fibres in isolated preparations. Linear frequency response analysis over a frequency range of 0.1--100Hz was performed under steady-state conditions, using small amplitude, sinusoidal water displacements produced by a glass sphere at a short distance from the skin. Period histograms of afferent nerve activity were computed, and amplitude, phase and mean activity of the response were determined by means of Fourier analysis. A standardization procedure at the start of each experiment made scaling of the frequency responses of different preparations unnecessary. The results show that for small stimulus amplitudes the response of the lateral-line organ over the whole range of frequencies studied can adequately be described as a modulation of the spontaneous activity. The amplitude of the response is proportional to the stimulus amplitude, and the phase of the response is independent of stimulus amplitude. The lateral-line organ of Xenopus laevis can thus be regarded as a linear system for stimuli which produce modulation of the spontaneous activity. The frequency response demonstrates unequivocally that the lateral-line organ of Xenopus laevis functions as a water velocity detector. For frequencies of stimulation from 0.1--20Hz the gain increases with a slope of 7.5 dB/oct, and up to 5Hz the response is almost in phase with the water velocity. The extent to which the different transmission steps between stimulus and response will contribute to the frequency response is discussed.

Effects of aldrin-transdiol on neuromuscular facilitation and depression.

The effects of aldrin-transdiol, one of the active metabolites of the insecticide dieldrin, on evoked transmitter release, neuromuscular facilitation and neuromuscular depression have been studied in frog sartorius nerve-muscle preparations. Conventional techniques of intracellular recordings were used. Aldrin-transdiol caused a marked increase in end-plate potential amplitude under conditions of low quantal content, i.e., in Mg2+-blocked junctions. The increase in end-plate potential amplitude was less pronounced in curarized junctions, in which the transmitter release was not impaired. Concomitant with the increase in end-plate potential size there was a marked enhancement of facilitation during short trains of stimuli applied to the motor nerve. The decay of facilitation was, however, not seriously affected by aldrin-transdiol. These effects may be explained in terms of the known 'calcium hypothesis' by assuring that aldrin-transdiol increases the amount of calcium which enters the nerve terminal during the nerve impulse. The increase in end-plate potential amplitude and in facilitation by aldrin-transdiol was transient. At later stages of poisoning, end-plate potential and facilitation decreased below control level and neuromuscular depression was significantly enhanced. This latter effect may be the result of a direct inhibitory effect of aldrin-transdiol on transmitter mobilization. As far as the fall in end-plate potential amplitude is concerned the known suppressive action of aldrin-transdiol on the chemical sensitivity of the postsynaptic membrane and on the nerve action potential probably plays a part as well. Finally, neuromuscular transmission was completely blocked by aldrin-transdiol.