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Study question How do human embryos implant in physiological conditions and develop beyond blastocyst stage? Summary answer Human embryos apply forces during invasion of the matrix, the mural trophectoderm undergoes a quick compaction leading to a fast radial expansion of polar side What is known already Human embryos can be cultured beyond blastocyst stage using supplements of animal origin, such as fetal bovine serum. However, two main issues limit our understanding of the implantation of the human embryo. First, human embryos do not express fluorescent proteins, severely limiting the access to advanced live-imaging tools. Second, the animal serums may not represent the physiological peri-implantation conditions of the human embryo, thus limiting the reproducibility of the in vitro experiments. Here we combine label-free multiphoton imaging with media formulations including clinical-grade protein supplements from human plasma unlocking the observation of the human embryo in 3D and physiological conditions Study design, size, duration We have cultured 150 human embryos from D3 until blastocyst stage and also more than 600 mouse embryos from zygote until blastocyst stage and then transferred them into a 3D implantation platform. All embryos were supplemented with clinical-grade human-derived serum, which is rich in globulins and growth factors, and commercial basal media. The implantation was monitored for 3-to-5 days up until day 11. Traction-force microscopy and molecular imaging was employed to quantify implantation markers. Participants/materials, setting, methods Implantation was measured using our novel version of traction-force microscopy, which reveals the force applied by entire embryos during live-imaging experiments. Multiphoton illumination of autofluorescent molecules allows to reconstruct images of live human embryos on 3D at unprecedented resolution. Key molecular markers for the formation of the epiblast rosette (OCT4), extra-embryonic visceral endoderm (GATA4), extra-embryonic ectoderm and trophoblast (CDX2) and primary yolk sac and pre-amniotic cavity were reconstructed in 3D using high resolution confocal imaging. Main results and the role of chance The implantation of human embryos was visualized in 3D movies at high resolution, during time-resolved experiments and compared to implantation in mouse embryos. Human embryos undergo a remarkable compaction upon attachment of the polar trophectoderm and thereafter they implant by engaging the matrix through many different points (>8). The post-implantation compaction leads to a strong invasion of the polar trophectoderm, digging large holes in the matrix and expanding radially. Some trophoblast cells escape the embryo colonizing the matrix. Human embryos expressed the correct pattern of molecular markers by day 11 of culture. Actin staining and OCT4 expression revealed the formation of a central rosette defining the pre-amniotic cavity. GATA6 expression defined the visceral endoderm and the formation of a primordial yolk sac. Mouse embryos conversely, compact upon implantation of the mural trophectoderm and are less invasive, typically anchoring to the matrix through 2-to-3 points. Post-implantation culture of mouse embryos starting at zygote was possible until the formation and large expansion of the amniotic cavity, which constituted 2/3 of the whole embryo size, corresponding with the initial steps of gastrulation, longer than any previous protocol Transfer of mouse blastocysts back to receptive mothers resulted in birth of viable litter. Limitations, reasons for caution This study has been performed with human embryos ex vivo using bioengineering technology to implant embryos outside the mother uterus. The bio-compatibility of our serum supplements was validated by live birth of transferred embryos in animal models, but would require further evaluation in clinical trials. Wider implications of the findings Our work stresses the safety and efficiency of supplementing embryo culture and particularly embryo transfer, with human plasma-derived serums of clinical-grade. The correct expression of main molecular markers indicates that this type of supplementation promotes implantation physiologically, which opens the possibility to employ these serums in the IVF laboratory. Trial registration number Not applicable

Study question Are fresh human sperm samples affected by different gravitational conditions than on Earth? Summary answer Motility and vitality of fresh human sperm samples are significantly decreased under microgravity conditions obtained by parabolic flight. What is known already Microgravity effects on the male reproductive system have mainly been studied in the animal model with diverse results and discouraging extrapolation in humans. While an increased motility was reported in bulls, mice models showed a decrease. Although preliminary data from the Micro-11 experiment presented by NASA reported human sperm alterations after microgravity exposure, our first study failed to show any significant effect of microgravity on “frozen” samples, suggesting that human sperm could be safely shipped outside the earth if important aspects related with cryopreservation were solved. Study design, size, duration Prospective study carried out in collaboration between the ART centre, a Technical University, and an Aviation Club specializing in parabolic flights.Two parabolic flights were conducted between 2020-2021, each consisting of 20 parabolic maneuvers, which means 160 seconds of microgravity exposure per sample. Fifteen sperm samples obtained from healthy men were included in the study in order to analyse the effects of microgravity and compare the results with those obtained in Earth gravity. Participants/materials, setting, methods Fresh sperm samples were checked pre-flight to evaluate vitality, concentration, motility and morphology. Samples were split into two to compare the effects of different gravity exposure: microgravity (flight) and Earth gravity (ground). After the flight, the same analysis were repeated, plus kinematics, DNA fragmentation by sperm chromatin dispersion, apoptosis by magnetic activated cell sorting, and oxidative stress by colorimetric test (Halosperm-Halotech). Computer Aided Semen Analysis (SCA-Scope) was used for cell counting. Main results and the role of chance On comparison of the mean values between fresh samples exposed to microgravity and those maintained on Earth gravity, statistical significant differences (p < 0,05) were found in the following parameters: vitality (69,7 ± 9,9 vs 72,4 ± 9,7 %), motile sperm concentration (23,7 ± 15,3 M/ml vs 31,5 ± 25,1 M/ml), grade “a” sperm concentration (8,7 ± 6,5 vs 11,7 ± 9,9 M/ml), percentage of spermatozoa with progressive motility (30 ± 12,9 vs 36 ± 14,3 %), curvilinear motility-VCL (45,7 ± 12,8 vs 47,7 ± 13,3 μm/s). Under the study conditions, non-statistically significant differences were observed in the other kinematic parameters: Lineal Velocity (VSL), Average Path Velocity (VAP), Straightness (STR), Amplitude of Lateral Head displacement (ALH), Linearity (LIN), Wobble (WOB), Beat-Cross Frequency (BCF), total sperm concentration (81,7 ± 112,1 vs 79,7 ± 89,8 M/ml), morphology (11,3 ± 6,3 vs 10,6 ± 5,3%), DNA fragmentation (14,6 ± 9,6 vs 15,7 ± 9,4), apoptosis (2,8 ± 2,8 vs 3,8 ± 4,4) and oxidative stress, since all samples maintained the same stress level in both splits. Limitations, reasons for caution Parabolic flight is an accepted ground-based method for obtaining microgravity conditions, but provides a short period of elapsed exposure to microgravity. Therefore, the results obtained need to be confirmed by using other platforms that provide a much longer time of exposure. More cases must be analysed to confirm the results. Wider implications of the findings Short exposure to microgravity significantly decreases sperm motility and vitality. Such an effect is likely to be stronger with longer exposure. These findings should be taken into account since this may eventually affect sperm fertilizing capacity and therefore natural conception or ART with fresh/frozen sperm, outside of the Earth. Trial registration number NCT03760783

From August 27–28, 2020 the Epilepsy Foundation hosted the Pipeline Conference, exploring emerging issues related to antiepileptic drug and device development. The conference featured epilepsy therapeutic companies and academic laboratories developing drugs for focal epilepsies, innovations for rare and ultra-rare diseases, and devices both in clinical trials and approved for use. In this paper, we outline the virtual presentations by the authors, including novel data from their development pipeline.

Objectives: Mild cognitive impairment (MCI) is associated with an increased risk of further cognitive decline, partly depending on demographics and biomarker status. The aim of the present study was to survey the clinical practices of physicians in terms of biomarker counseling, management, and follow-up in European expert centers diagnosing patients with MCI. Methods: An online email survey was distributed to physicians affiliated with European Alzheimer's disease Consortium centers (Northern Europe: 10 centers; Eastern and Central Europe: 9 centers; and Southern Europe: 15 centers) with questions on attitudes toward biomarkers and biomarker counseling in MCI and dementia. This included postbiomarker counseling and the process of diagnostic disclosure of MCI, as well as treatment and follow-up in MCI. Results: The response rate for the survey was 80.9% (34 of 42 centers) across 20 countries. A large majority of physicians had access to biomarkers and found them useful. Pre- and postbiomarker counseling varied across centers, as did practices for referral to support groups and advice on preventive strategies. Less than half reported discussing driving and advance care planning with patients with MCI. Conclusions: The variability in clinical practices across centers calls for better biomarker counseling and better training to improve communication skills. Future initiatives should address the importance of communicating preventive strategies and advance planning. © 2020 John Wiley & Sons Ltd.

On May 22-24, 2019, the 15th Antiepileptic Drug and Device (AEDD) Trials Conference was held, which focused on current issues related to AEDD development from preclinical models to clinical prognostication. The conference featured regulatory agencies, academic laboratories, and healthcare companies involved in emerging epilepsy therapies and research. The program included discussions around funding and support for investigations in epilepsy and neurologic research, clinical trial design and integrated outcome measures for people with epilepsy, and drug development and upcoming disease-modifying therapies. Finally, the conference included updates from the preclinical, clinical, and device pipeline. Summaries of the talks are provided in this paper, with the various pipeline therapeutics in the listed tables to be outlined in a subsequent publication.

This letter presents an insightful and significant improvement of the cylindrical cavity model and an experimental methodology to compensate the mechanical bending effects on cylindrical microstrip antennas. In the first part of this letter, by studying analytical and numerical results from a nonradiating resonant rectangular cavity, a new function for the resonant length of the circumferential mode that is governed by the antenna thickness and cylinder radius was determined. In the second part, an experimental methodology is presented to compensate the copper elongation effect due to bending. To validate the proposed techniques, a circularly polarized rectangular microstrip antenna was designed and built to operate at 2.2 GHz with an input impedance of 50 Ω for a metallic cylinder with a radius of 25 cm. The experimental results showed that the new approaches to calculate the circumferential resonant frequency and elongation effect due to bending improved the cylindrical antenna design significantly.

Introduction: Large variability among Alzheimer's disease (AD) cases might impact genetic discoveries and complicate dissection of underlying biological pathways. Methods: Genome Research at Fundacio ACE (GR@ACE) is a genome-wide study of dementia and its clinical endophenotypes, defined based on AD's clinical certainty and vascular burden. We assessed the impact of known AD loci across endophenotypes to generate loci categories. We incorporated gene coexpression data and conducted pathway analysis per category. Finally, to evaluate the effect of heterogeneity in genetic studies, GR@ACE series were meta-analyzed with additional genome-wide association study data sets. Results: We classified known AD loci into three categories, which might reflect the disease clinical heterogeneity. Vascular processes were only detected as a causal mechanism in probable AD. The meta-analysis strategy revealed the ANKRD31-rs4704171 and NDUFAF6-rs10098778 and confirmed SCIMP-rs7225151 and CD33-rs3865444. Discussion: The regulation of vasculature is a prominent causal component of probable AD. GR@ACE meta-analysis revealed novel AD genetic signals, strongly driven by the presence of clinical heterogeneity in the AD series., The authors would like to thank patients and controls who participated in this project. The Genome Research @ Fundacio ACE project (GR@ACE) is supported by Fundacion bancaria La Caixa, Grifols SA, Fundacio ACE, and ISCIII (Ministry of Health, Spain). They also want to thank the private sponsors who support the basic and clinical projects of our institution (Piramal AG, Laboratorios Echevarne, Araclon Biotech S.A., and Fundacio ACE). They are indebted to the Trinitat Port-Carbo legacy and her family for their support of Fundacio ACE research programs. Fundacio ACE is a participating center in the Dementia Genetics Spanish Consortium (DEGESCO). A.R. and M.B. receive support from the European Union/EFPIA Innovative Medicines Initiative Joint undertaking ADAPTED and MOPEAD projects (grant numbers 115975 and 115985, respectively). M.B. and A.R. are also supported by national grants PI13/02434, PI16/01861, and PI17/01474. Accion Estrategica en Salud is integrated into the Spanish National R + D + I Plan and funded by ISCIII (Instituto de Salud Carlos III)-Subdireccion General de Evaluacion and the Fondo Europeo de Desarrollo Regional (FEDER-Una manera de Hacer Europa). L.M.R. is supported by Consejeria de Salud de la Junta de Andalucia (grant PI-0001/2017). Control samples and data from patients included in this study were provided in part by the National DNA Bank Carlos III (www.bancoadn.org, University of Salamanca, Spain) and Hospital Universitario Virgen de Valme (Sevilla, Spain); they were processed after standard operating procedures with the appropriate approval of the Ethical and Scientific Committee. The present work was performed as part of the Biochemistry, Molecular Biology, and Biomedicine doctoral program of S. MorenoGrau at Universitat Autonoma de Barcelona (Barcelona, Spain).r Data collection and sharing for this project was partially funded by the Alzheimer's Disease Neuroimaging Initiative (ADNI) (National Institutes of Health grant U01 AG024904) and DOD ADNI (Department of Defense award number W81XWH-12-2-0012). The ADNI is funded by the National Institute on Aging and the National Institute of Biomedical Imaging and Bioengineering, as well as through generous contributions from the following: AbbVie; the Alzheimer's Association; the Alzheimer's Drug Discovery Foundation; Araclon Biotech; BioClinica, Inc.; Biogen; Bristol-Myers Squibb Company; CereSpir, Inc.; Cogstate; Eisai Inc.; Elan Pharmaceuticals, Inc.; Eli Lilly and Company; EuroImmun; F. Hoffmann-La Roche Ltd and its affiliated company Genentech, Inc.; Fujirebio; GE Healthcare; IXICO Ltd.; Janssen Alzheimer Immunotherapy Research & Development, LLC.; Johnson & Johnson Pharmaceutical Research & Development LLC.; Lumosity; Lundbeck; Merck & Co., Inc.; Meso Scale Diagnostics, LLC.; NeuroRx Research; Neurotrack Technologies; Novartis Pharmaceuticals Corporation; Pfizer Inc.; Piramal Imaging; Servier; Takeda Pharmaceutical Company; and Transition Therapeutics. The Canadian Institutes of Health Research provides funds to support ADNI clinical sites in Canada. Private sector contributions are facilitated by the Foundation for the National Institutes of Health (www.fnih.org). The grantee organization is the Northern California Institute for Research and Education, and the study was coordinated by the Alzheimer's Therapeutic Research Institute at the University of Southern California. ADNI data are disseminated by the Laboratory for NeuroImaging at the University of Southern California.; r The AddNeuroMed data are from a public-private partnership supported by EFPIA companies and SMEs as part of InnoMed (Innovative Medicines in Europe), an integrated project funded by the European Union of the Sixth Framework program priority FP6-2004-LIFESCIHEALTH-5. Clinical leads responsible for data collection are Iwona K1oszewska (Lodz), Simon Lovestone (London), Patrizia Mecocci (Perugia), Hilkka Soininen (Kuopio), Magda Tsolaki (Thessaloniki), and Bruno Vellas (Toulouse). Imaging leads are Andy Simmons (London), Lars-Olad Wahlund (Stockholm), and Christian Spenger (Zurich). Bioinformatics leads are Richard Dobson (London) and Stephen Newhouse (London).r Funding support for the Alzheimer's Disease Genetics Consortium (ADGC) was provided through the NIA Division of Neuroscience (U01-AG032984).r The Mayo Clinic Alzheimer's Disease Genetic Studies, led by Dr. Nilufer Ertekin-Taner and Dr. Steven G. Younkin at the Mayo Clinic in Jacksonville, FL, used samples from the Mayo Clinic Study of Aging, the Mayo Clinic Alzheimer's Disease Research Center, and the Mayo Clinic Brain Bank. Data collection was supported through funding by NIA grants P50 AG016574, R01 AG032990, U01 AG046139, R01 AG018023, U01 AG006576, U01 AG006786, R01 AG025711, R01 AG017216, and R01 AG003949, NINDS grant R01 NS080820, the CurePSP Foundation, and support from the Mayo Foundation.r The Neocodex-Murcia study was funded by the Fundacion Alzheimur (Murcia), the Ministerio de Educacion y Ciencia (Gobierno de Espana), Corporacion Tecnologica de Andalucia, Agencia IDEA (Consejeria de Innovacion, Junta de Andalucia), the Diabetes Research Laboratory, and the Biomedical Research Foundation. University Hospital Clinico San Carlos has been supported by CIBER de Diabetes y Enfermedades Metabolicas Asociadas (CIBERDEM); CIBERDEM is an ISCIII Project.r The ROS/MAP study data were provided by the Rush Alzheimer's Disease Center, Rush University Medical Center, Chicago. Data collection was supported through funding by NIA grants P30AG10161, R01AG15819, R01AG17917, R01AG30146, R01AG36836, U01AG32984 and U01AG46152, the Illinois Department of Public Health, and the Translational Genomics Research Institute.r The TGEN study was supported by Kronos Life Science Laboratories, the National Institute on Aging (Arizona Alzheimer's Disease Center grants P30 AG19610 and RO1 AG023193, the Mayo Clinic Alzheimer's Disease Center grant P50 AG16574, and the Intramural Research Program), the National Alzheimer's Coordinating Center (U01 AG016976), and the state of Arizona.r The authors thank the International Genomics of Alzheimer's Project (IGAP) for providing summary result data for these analyses. The investigators within IGAP contributed to the design and implementation of IGAP and/or provided data but did not participate in analysis or writing of this report. IGAP was made possible by the generous participation of the control subjects, the patients, and their families. The i-Select chip was funded by the French National Foundation on Alzheimer's disease and related disorders. European Alzheimer's Disease Initiative (EADI) was supported by the LABEX (laboratory of excellence program investment for the future) DISTALZ grant, Inserm, Institut Pasteur de Lille, Universite de Lille 2, and the Lille University Hospital. GERAD was supported by the Medical Research Council (grant no. 503480), Alzheimer's Research UK (grant no. 503176), the Wellcome Trust (grant no. 082604/2/07/Z), and German Federal Ministry of Education and Research (BMBF): Competence Network Dementia (CND) grant no.; 01GI0102, 01GI0711, 01GI0420. CHARGE was partly supported by the NIH/NIA grant R01 AG033193 and the NIA AG081220 and AGES contract N01-AG12100, the NHLBI grant R01 HL105756, the Icelandic Heart Association, and the Erasmus Medical Center and Erasmus University. ADGC was supported by the NIH/NIA grants: U01 AG032984, U24 AG021886, U01 AG016976, and the Alzheimer's Association grant ADGC-10-196728.

This paper presents a new cavity model surrogate-based optimization for electrically thick probe-fed circularly polarized rectangular microstrip antennas. The proposed methodology feeds back input impedance and far-field information from full-wave electromagnetic solvers in order to calibrate known sources of inaccuracy within the cavity model. Said calibration is achieved by solving a constrained non-linear least squares problem. The calibrated cavity model is used to generate a new antenna geometry that will be subjected to the same calibration process until convergence is achieved. The amount of iterations needed to achieve convergence for several configurations is less than or equal to three, this indicates that the cavity model surrogate-based optimization can be used as an efficient design technique. In addition, the optimization process correctly anticipated that unequal fringing field lengths for the resonant modes are required to calibrate the cavity model, demonstrating that great physical insight is gained throughout the optimization process. For the validation of the proposed design procedure, eight electrically thick antennas were designed and optimized, of which three were manufactured and tested. Good agreement between simulated and experimental results were observed, validating the applicability of the proposed strategy.

Table S2. Exploratory analysis. (DOCX 23 kb)

Figure S4. APOE and plasma Aβ ratios. The effects of APOE genotype on plasma Aβ levels using ANOVA between APOE ε4 carriers and noncarriers in a boxplot representation with outlier analysis. (PDF 93 kb)

Table S3. Regression analyses between Aβ plasma ratios and FBB-PET SUVR. (DOCX 18 kb)

Table S5. Interaction between APOE and L_TP42/40. (DOCX 14 kb)

Figure S2. Scatter plots for FBB-PET global SUVR and Aβ plasma ratios in SCD subjects. Correlations between plasma biomarkers and brain Aβ burden. Biomarkers values plotted against SUVR values from FBB-PET imaging: FP42/40 (A), BP42/40 (B), FP42/TP42 (C), and FP40/TP40 (D). (PDF 286 kb)

Figure S3. Linear regression between FBB-PET and Aβ TP42/40 plasma ratio in APOE ε4 stratification SCD population. A) APOE ε4 carriers; B) APOE ε4 noncarriers. (PDF 115 kb)

Figure S1. A) Distribution of FBB-PET and plasma ratios. B) Shapiro-Wilk test for FBB-PET and plasma ratios. C) Log distributions FBB-PET and plasma ratios. D) Shapiro-Wilk test for logarithmic FBB-PET and log-plasma ratios. A, B) Distributions and Shapiro-Wilk test for plasma ratios and FBB-PET to test normality. C, D) Distributions and Shapiro-Wilk test for transformed to logarithmic plasma ratios and FBB-PET to test normality. (PDF 299 kb)

Table S4. ANOVAs comparing APOE ε4 carriers vs noncarriers. (DOCX 14 kb)

Introduction Subjective cognitive decline (SCD) has been proposed as a potential preclinical stage of Alzheimer's disease (AD). Nevertheless, the genetic and biomarker profiles of SCD individuals remain mostly unexplored. Methods We evaluated apolipoprotein E ( APOE ) e4's effect in the risk of presenting SCD, using the Fundacio ACE Healthy Brain Initiative (FACEHBI) SCD cohort and Spanish controls, and performed a meta-analysis addressing the same question. We assessed the relationship between APOE dosage and brain amyloid burden in the FACEHBI SCD and Alzheimer's Disease Neuroimaging Initiative cohorts. Results Analysis of the FACEHBI cohort and the meta-analysis demonstrated SCD individuals presented higher allelic frequencies of APOE e4 with respect to controls. APOE dosage explained 9% (FACEHBI cohort) and 11% (FACEHBI and Alzheimer's Disease Neuroimaging Initiative cohorts) of the variance of cerebral amyloid levels. Discussion The FACEHBI sample presents APOE e4 enrichment, suggesting that a pool of AD patients is nested in our sample. Cerebral amyloid levels are partially explained by the APOE allele dosage, suggesting that other genetic or epigenetic factors are involved in this AD endophenotype.