To examine real-world effectiveness of ubrogepant for the acute treatment of migraine in patients who switched to ubrogepant due to the lack of efficacy of their prior acute treatment.

The purpose of this chapter is to review some of the factors that influence laboratory and clinical outcomes, broadly under the heading of optimal handling techniques. Two principal environments are encountered – inside and outside the incubator. The chapter will address media buffers, gas atmosphere, timing and setting up culture or holding vessels, protection of medium performance, temperature relative to handling gametes and embryos, lighting, pH, incubation choices, and workflow. The importance of the interplay between these variables cannot be overlooked.

Research question: What are the viscosities of media used for human embryo transfer and what is the possible effect of viscosity as it relates to interactions between transfer media and uterine fluid. Design: Chamber slide filling times, in seconds, were used to calculate viscosity for each commercial and in-house modified medium, with 12 or 24 replicates per medium under standard operating procedure temperature and gas equilibration conditions used for embryo transfer. Means, standard deviations and coefficients of variation were calculated, and each viscosity was estimated using a regression equation; viscosities for each medium were presented for comparative purposes. Results: Complete culture media (G1-Plus, G2-Plus, G-TL, 1-Step, Global Total, Global Total HEPES, and Sperm Wash Medium) had viscosity estimates of 1.65 cP, 1.77 cP, 1.68 cP, 1.29cP, 1.18 cP, 1.15 cP, and 1.20 cP, respectively. Complete transfer media (EmbryoGlue, UTM), had viscosity estimates of 3.59 cP and 1.28 cP, respectively. Global HEPES medium with 10%, 20%, 30%, and 50% synthetic serum substitute (SSS) volume per volume had viscosity estimates 1.16 cP, 1.23 cP, 1.25 cP, and 1.34 cP, respectively. For reference, water had a viscosity estimate of 1.06 cP. Conclusions: A relatively narrow distribution of viscosities was observed across several transfer media despite the various commercial or in-house modifications. These data demonstrate the vast difference between viscosities of embryo transfer media and the assumed viscosity of uterine fluid (1000 cP). Contemporary embryo transfer media may be well-suited for IVF, but evaluation of all variables, e.g. media viscosity in the context of embryo transfer, adds to the knowledge base that should be available to practitioners.

To determine liquid nitrogen evaporation rates of intact liquid nitrogen storage tanks and tanks with their vacuum removed. Donated storage tank performance (LN2 evaporation) was evaluated before and after induced vacuum failure. Vacuum of each tank was removed by drilling through the vacuum port. Temperature probes were placed 2 in. below the bottom of the styrofoam cap/plug, and tanks were weighed every 3 h. Evaporation rate and time from failure to the critical temperature was determined. Storage tanks with failed vacuum have a much higher evaporation rate than those with intact vacuum; evaporation rates increased dramatically within 3 to 6 h in the smaller tanks, and time to complete depletion varied according to starting LN2 volume. Tanks with storage racks/specimens may have altered evaporation profiles compared to tanks without. Locating temperature probes 2 in. below the styrofoam cap/plug suggests that for most applications, alarms would sound approximately 1 h prior to reaching the critical warming temperature, approximately − 130 °C. External signs of vacuum loss were dramatic: vapor, frost, and audible movement of air. For the first time, we have data on how liquid nitrogen storage tanks behave when their vacuum is removed. These findings are conservative; each lab must consider starting volume, tank size/capacity, function (storage or shipping), age, and pre-existing evaporation behavior in order to develop an emergency response to critical tank failure. Times to complete failure/evaporation and critical warming temperature after vacuum loss are different; these data should be considered when evaluating tank alarm systems.

Experimental results from six prototype devices demonstrate that pressure changes induced in a liquid bridge via electrowetting can generate large deflections (20-75 mu m) of an elastomeric membrane similar to those used in lab-on-a-chip microfluidic devices. In all cases deflections are obtained with a low voltage (20 V) and very small power consumption (< 1 mu W). The effects of variations in the bridge size and membrane dimensions on measured displacements are examined. Theoretical predictions are in good agreement with the measured displacements in those cases where the liquid contact angles could be measured within the devices during electrowetting. Contact angle hysteresis and charge injection into the dielectric layers limited the repeatability of deflection behavior during repeated cycling. Approaches for achieving greater deflections and improved repeatability are discussed.

Limitations to embryo culture techniques in the early years of human in vitro fertilization (IVF) required transfer and/or cryopreservation of early cleavage-stage embryos on day 2 and day 3. Embryo quality scoring systems for these early cleavage-stage embryos were simplistic as the number of blastomeres, evenness of blastomere size, placement of blastomeres and extent of fragmentation were very evident. Extended culture of embryos to the blastocyst stage effectively doubled the time these embryos were maintained in vitro. Observations were commonly made at fertilization determination, mid-culture (day 3) and again at the end of the culture period (days 5 and 6). Evaluations of embryos on days 2 and 4 were then skipped in many laboratories. Qualitative selection protocols for morula-stage embryos rely on metrics associated with timing of compaction, extent of compaction, morphology of blastomeres and nuclei, extent of fragmentation and cavitation, if present. This chapter will touch on developmental details integrated into the suggested grading protocol, as well as providing evidence that morulae are overlooked as a viable clinical choice for transfer and for cryopreservation.

Low-voltage electrowetting devices allow significant contact angle changes below a 50 V bias; however, operation under prolonged cycling and failure modes have not yet been sufficiently elucidated. In this work, the failure modes and performance degradation of Cytop (23-210 nm)/aluminum oxide (15-44 nm) bilayers have been investigated. Contact angle and leakage current were measured during stepped voltage measurements up to failure, showing three electrowetting response regimes: ideal Young-Lippmann behavior, contact angle saturation, and dielectric breakdown. The onset of ionic conduction in aluminum oxide and the resulting breakdown control when the layer would ultimately fail, but the thickness of the Cytop layer determined the achievable contact angle versus voltage characteristics. Cyclic electrowetting measurements studied the repeatability of contact angle change using an applied voltage above or below the voltage drop needed for polymer breakdown (VT). Results show repeatable electrowetting below VT and a rapidly diminishing contact angle response above VT. The leakage current and injected charge cannot be used to comprehensively assess the stability of the system during operation. The contact potential difference measured with a Kelvin probe provides an alternative means of assessing the extent of the damage.

Purpose To evaluate the transition from a proven slow-cooling cryopreservation method to a commercial large-volume vitrification system for human blastocysts. Methods Retrospective analysis of de-identified laboratory and clinical data from January 2012 to present date for all frozen embryo replacement (FET) cycles was undertaken. Cryopreservation of trophectoderm-biopsied or non-biopsied blastocysts utilized during this time period was logged as either slow-cooling, small-volume vitrification, or large-volume vitrification. Blastocyst survival post-warm or post-thaw, clinical pregnancy following FET, and implantation rates were identified for each respective cryopreservation method. Results Embryo survival was highest for large-volume vitrification compared to micro-volume vitrification and slow-cooling; 187/193 (96.9 %), 27/32 (84.4 %), and 244/272 (89.7 %), respectively. Survival of biopsied and non-biopsied blastocysts vitrified using the large-volume system was 105/109 (96.3 %) and 82/84 (97.6 %), respectively. Survival for micro-volume biopsied and non-biopsied blastocysts was 16/30 (83.3 %) and 2/2 (100.0 %) respectively. Slow-cooling post-thaw embryo survival was 272/244 (89.7 %). Clinical pregnancy and implantation rates outcomes for non-biopsied embryos were similar between large-volume and slow-cooling cryopreservation methods, 18/39 (46.2 %) clinical pregnancy and 24/82 (29.3 %) implantation/embryo, and 52/116 (44.8 %) clinical pregnancy and 67/244 (27.5 %) implantation/embryo, respectively. Comparing outcomes for biopsied embryos, clinical pregnancy and implantation rates were 39/67 (58.2 %) clinical pregnancy and 50/105 (47.6 %) implantation/embryo and 4/16 (25 %) clinical pregnancy and 6/25 (24.0 %) implantation/embryo, respectively. Conclusions The LifeGlobal large-volume vitrification system proved to be very reliable, simple to learn and implement in the laboratory. Clinically large-volume vitrification was as, or more effective compared to slow-cooling cryopreservation in terms of recovery of viable embryos in this laboratory.

The purpose of this study was to quantitate changes in seminal volume, sperm count, motility, qualitative forward progression, and total motile sperm cells per ejaculate, across three consecutive ejaculates collected from individuals within 24 h preceding an IVF cycle.

We discuss the electrochemical formation of metal oxide films as candidate materials for the primary dielectric layer in electrowetting-on-dielectric (EWOD) systems. The ability to produce significant contact angle reduction at low voltage requires thin (<100nm), high dielectric strength films that also exhibit a high breakdown field; these requirements are met in a wide variety of metal oxides. A detailed understanding of materials as well as electronic and ionic transport in dielectrics are essential for the development and performance enhancement of these devices; we describe the process of oxides in film formation, the peculiar defect structure, electrical transport processes, as well as degradation and breakdown. Additionally, examples of low-voltage EWOD systems based on oxide/hydrophobic bilayers are discussed along with current efforts to improve performance of the metal oxide layers.

Several parameters associated with the configuration of a microliter aqueous droplet are determined by the electrical impedance measurements conducted during electrowetting. In each case, the wetted area of a dielectric-coated electrode is first determined from the impedance data, and the known dielectric thickness and permittivity. The wetted area data is then employed in a series of identification problems. In the simplest of these, we demonstrate that the contact angle can be accurately found as a function of applied bias voltage. Thus, the Lippmann-Young (LY) curve of an electrowetting system can be determined from the impedance data alone without the use of any optical measurements. In another test, we show that the volume of a microliter droplet can be estimated from electrical impedance measurements with an error of <;1%. A similar algorithm was employed to determine the surface tension of the interface between the droplet and the surrounding oil to an accuracy of 4%. In still another identification experiment, the impedance data from multiple bias voltages were used as a batch to identify several unknown configuration parameters simultaneously (volume, interfacial tension, and zero-voltage contact angle). It is shown that identification accuracy is degraded in this case, because there exists a direction within the parameter space of low error sensitivity. Determining multiple parameters together, therefore, requires that the ideal LY relation fits the electrowetting behavior well over the range of voltages employed and that the dielectric parameters are accurately known.

Using impedance spectroscopy, we have determined models for the elements which determine the ac electrical behavior in electrowetting on dielectric (EWOD) systems. Three commonly used EWOD electrode configurations were analyzed. In each case, the impedance can be modeled by a combination of elements, including the solution resistance, the capacitance of the dielectric layer, and the constant phase impedance of the electrode double layers. The sensitivity of the system's impedance to variations in the electrowetted area is also analyzed for these common configurations. We also demonstrate that the impedance per unit area of typical EWOD systems is invariant to bias voltage.

In oviparous animals, the egg hatches outside of the body and is exposed to light; in some cases throughout the development of the fetus. In mammals, fertilization and the growth of embryos in vivo occurs in the dark but in human IVF, these embryos are exposed to variable light sources and intensities. Light can affect embryonic development in some species via either a direct toxic effect on the embryo, or indirectly via photo-oxidation of components in the media or oil. Although data regarding the effect of light on human embryos is lacking, it is prudent to take appropriate steps to minimize the potential harmful effects of both ambient and microscopic light on embryos.

Octadecylphosphonic acid reacts with oxidized aluminum surfaces to form a self-assembled monolayer film. The effectiveness of the monolayers are examined for use in an electrowetting-on-dielectric application using anodized aluminum as the substrate. We report growth of the monolayers by monitoring contact angle and SEM imaging. Electrowetting is characterized in static and dynamic configuration, giving advancing, receding, and contact hysteresis. The films greatly reduce the voltage requirement for drop movement, but the high hysteresis currently limits the effectiveness at low voltage operation.

The electronic and ionic transports in 32–56 nm thick anodic aluminum oxide films are investigated before and after a 1-h anneal at 200–400 °C in argon. Results are correlated to their defect density as measured by the Mott-Schottky technique. Solid state measurements show that electronic conduction upon annealing is hindered by an increase in the Schottky emission barrier, induced by a reduction in dopant density. Using an electrochemical contact, the films fail rapidly under cathodic polarization, unless defect density is decreased down to 1017 cm−3, resulting in a three order of magnitude reduction in current and no visible gas evolution. Under anodic polarization, the decrease in defect density delays the onset of ionic conduction as well as further oxide growth and failure.

ObjectivePatients want alternatives to routine semen analysis (SA). Sperm Check® Fertility (kit) in drugstores is FDA approved for home use to screen for sperm. Kit result is low or normal.; 20m/ml normal value based on SP-10 antibody reaction.DesignThe kit was tested on 40 patients at our lab for routine SA; actual and expected (using SA results) were compared.Materials and MethodsKit has sample cup, syringe (100ul), lysis buffer (2ml), test cassette. After 20 min, 100ul semen added to buffer, mixed, incubated for 2 min. 6 drops added to cassette, results read at 7 min. A sensitivity curve (10, 15, 20, 25, 30 m/ml, in duplicate) was performed to test color development. Curve specimens; prepared from semen diluted with cleared seminal fluid. Manual SA was performed with 20ul chamber slides along with the kit for each patient.ResultsTabled 1Mean SA and selected pt. resultsn=40Volume mlCount m/mlMotility %Progression (1-4)Kit resultExpected resultMean, sd2.9, 1.560.9, 52.244.7, 22.31.9, 0.6Pt 30.2510.910.01.5NormalLowPt 105.08.066.71.5NormalLowPt 223.020.317.52.0LowNormal Open table in a new tab ConclusionKits performed as advertised. Limitations: 1) patients misdiagnosed may not seek medical advice, 2) the kit cannot tell low vs zero sperm count, 3) motility cannot be evaluated; e.g. no, or low motility, complete agglutination will go undetected, 4) total motile count, ejaculate volume issues are not considered. Recommendations: 1) patients with infertility or other concerns should not rely on the kit for diagnosis, 2) but if used, repeat 2 weeks apart for accuracy and talk to a provider, 3) consider the time and cost of routine SA vs kit limitations. ObjectivePatients want alternatives to routine semen analysis (SA). Sperm Check® Fertility (kit) in drugstores is FDA approved for home use to screen for sperm. Kit result is low or normal.; 20m/ml normal value based on SP-10 antibody reaction. Patients want alternatives to routine semen analysis (SA). Sperm Check® Fertility (kit) in drugstores is FDA approved for home use to screen for sperm. Kit result is low or normal.; 20m/ml normal value based on SP-10 antibody reaction. DesignThe kit was tested on 40 patients at our lab for routine SA; actual and expected (using SA results) were compared. The kit was tested on 40 patients at our lab for routine SA; actual and expected (using SA results) were compared. Materials and MethodsKit has sample cup, syringe (100ul), lysis buffer (2ml), test cassette. After 20 min, 100ul semen added to buffer, mixed, incubated for 2 min. 6 drops added to cassette, results read at 7 min. A sensitivity curve (10, 15, 20, 25, 30 m/ml, in duplicate) was performed to test color development. Curve specimens; prepared from semen diluted with cleared seminal fluid. Manual SA was performed with 20ul chamber slides along with the kit for each patient. Kit has sample cup, syringe (100ul), lysis buffer (2ml), test cassette. After 20 min, 100ul semen added to buffer, mixed, incubated for 2 min. 6 drops added to cassette, results read at 7 min. A sensitivity curve (10, 15, 20, 25, 30 m/ml, in duplicate) was performed to test color development. Curve specimens; prepared from semen diluted with cleared seminal fluid. Manual SA was performed with 20ul chamber slides along with the kit for each patient. ResultsTabled 1Mean SA and selected pt. resultsn=40Volume mlCount m/mlMotility %Progression (1-4)Kit resultExpected resultMean, sd2.9, 1.560.9, 52.244.7, 22.31.9, 0.6Pt 30.2510.910.01.5NormalLowPt 105.08.066.71.5NormalLowPt 223.020.317.52.0LowNormal Open table in a new tab ConclusionKits performed as advertised. Limitations: 1) patients misdiagnosed may not seek medical advice, 2) the kit cannot tell low vs zero sperm count, 3) motility cannot be evaluated; e.g. no, or low motility, complete agglutination will go undetected, 4) total motile count, ejaculate volume issues are not considered. Recommendations: 1) patients with infertility or other concerns should not rely on the kit for diagnosis, 2) but if used, repeat 2 weeks apart for accuracy and talk to a provider, 3) consider the time and cost of routine SA vs kit limitations. Kits performed as advertised. Limitations: 1) patients misdiagnosed may not seek medical advice, 2) the kit cannot tell low vs zero sperm count, 3) motility cannot be evaluated; e.g. no, or low motility, complete agglutination will go undetected, 4) total motile count, ejaculate volume issues are not considered. Recommendations: 1) patients with infertility or other concerns should not rely on the kit for diagnosis, 2) but if used, repeat 2 weeks apart for accuracy and talk to a provider, 3) consider the time and cost of routine SA vs kit limitations.

We study electrical properties and breakdown phenomena in metal/aluminum oxide/metal and electrolyte/aluminum oxide/metal contacts, with the aim to achieve a better understanding of failure modes and improve the performance of model electrowetting systems. Electrical conduction in anodic aluminum oxide dielectrics is dominated by the presence of electrically active trapping sites, resulting in various conduction mechanisms being dominant within distinct voltage ranges until hard breakdown occurs. Breakdown voltage depends on its polarity, due to the formation of a p-i-n junction within the oxide; such asymmetric behavior tends to disappear at larger oxide thickness. Electrolyte/dielectric contacts present an even more pronounced asymmetry in breakdown characteristics: a cathodic bias results in breakdown at low voltage, while under anodic bias high field ionic conduction starts before breakdown occurs. These phenomena are interpreted in terms of electrochemical reactions occurring at the surface: cathodic processes contribute to oxide dissolution and failure, while anodic processes result in additional oxide growth before breakdown.