O-086 End-to-end deep learning for recognition of ploidy status using time-lapse videos

Study question Our Retrospective study is to investigate an end-to-end deep learning model in identifying ploidy status through raw time-lapse video. Summary answer Our deep learning model demonstrates a proof of concept and potential in recognizing the ploidy status. What is known already Since the time-lapse system has been introduced into the IVF lab, the relationship between morphogenetic and ploidy status has been often discussed. However, the result has not yet reached a united conclusion due to some limitations such as human labeling. Besides the statistical approach, deep learning models have been utilized for ploidy prediction. As such approaches are single image-based, the performance remains unpromising as previous statistical-based research. Therefore, in order to move further toward clinical application, better research design and approach are needed. Study design, size, duration A retrospective analysis of the time-lapse videos and chromosomal status from 690 biopsied blastocysts cultured in a time-lapse incubator (EmbryoScope+, Vitrolife) between January 2017 and August 2018 in the Lee Women’s Hospital were assessed. The ploidy status of the blastocyst was derived from the PGT-A using high-resolution next-generation sequencing (hr-NGS). Embryo videos were obtained after normal fertilization through the intracytoplasmic sperm injection or conventional insemination. Participants/materials, setting, methods By randomly dividing the data into 80% and 20%, we developed our deep learning model based on Two-Stream Inflated 3D ConvNets(I3D) network. This model was trained by the 80% time-lapse videos and the PGT-A result. The remaining 20% has been tested by feeding the time-lapse video as input and the PGT-A prediction as output. Ploidy status was classified as Group 1 (aneuploidy) and Group 2 (euploidy and mosaicism). Main results and the role of chance Time-lapse videos were divided into 3-time partitions: day 1, day 1 to 3, and day 1 to 5. Deep learning models have been fed by RGB and optical flow. Combining 3 different time partitions with RGB, optical flow, and fused result from RGB and optical flow, we received nine sets of test results. According to the results, the longest time partition with the fusion method has the highest AUC result as 0.74, which appeared higher than the other eight experimental settings with a maximum increase of 0.17. Limitations, reasons for caution The present study is retrospective and future prospective research would help us to identify more key factors and improve this model. In addition, expanding sample size combined with cross-centered validation will also be considered in our future approach. Wider implications of the findings Group 1 and Group 2 approach provided deselection of aneuploidy embryos, while future deep learning approaches toward high mosaicism, low mosaicism, and euploidy will be needed, in order to provide a better clinical application. Trial registration number CS18082

Ibrutinib Off-Target Inhibition Inhibits Antibody-Dependent Cellular Phagocytosis but Not Efferocytosis of CLL Cells

Background Combinations of different targeted therapies, including Bruton tyrosine kinase (BTK) inhibitors and anti-CD20 monoclonal antibodies (mAbs) could improve treatment for CLL. Unexpectedly, the combination of ibrutinib (IBR) with rituximab did not show additional clinical benefit. However, IBR inhibits many off-target molecules that may limit therapeutic mAb clinical effectiveness and a more selective BTK inhibitor, such as acalabrutinib (ACALA), could be more effective in combination with mAb therapy. Initial data from the ELEVATE TN trial support this possibility. IBR off-target effects on antibody-dependent cellular phagocytosis (ADCP), the major mechanism of therapeutic mAb activity could explain this difference. Additionally, IBR induces a higher and longer duration increase in circulating lymphocytes than ACALA. IBR off-target effects on efferocytosis, another phagocytic process involved in apoptotic cell removal, might explain this difference. Methods Using state-of-the-art direct kinetic measurements of phagocytosis by time-lapse video, (Chu et al. J Cell Sci 2020;133:jcs237883) we investigated the effects of IBR and ACALA on phagocytosis (ADCP or efferocytosis) by human monocyte-derived macrophages (hMDM) in vitro. Live cell time-lapse video of 10 μg/ml rituximab (Genentech) mediated ADCP of CLL cells by CellTracker Deep Red (CTDR, Thermofisher) labeled hMDM (20:1 CLL:hMDM cell ratio) either untreated or treated with IBR or ACALA (3-fold serial dilutions from 100 to 0.41 μM) was imaged in a stage-top environmental chamber (37°C and 5% CO2) mounted onto a Nikon Ti-Eclipse inverted microscope with an ELWD 20x/0.45NA S Plan Fluor Ph1 objective and an Andor Zyla 5.5 sCMOS camera. Images were captured sequentially every 4 min over 2.8 h. For each experiment (n = 18), duplicate or triplicate wells for each drug concentration were imaged. For efferocytosis, live cell time-lapse video imaging of phagocytosis of pHrodo iFL Red STP ester (pHrodo Red, Thermo Fisher Scientific) labeled apoptotic CLL cells by CTDR-labeled hMDM (20:1 CLL:hMDM cell ratio) either untreated or treated with IBR or ACALA (2-fold serial dilutions from 10 to 1.25 μM) was collected every 4 min over 2.8 h. For each experiment (n = 7), duplicate or triplicate wells for each drug concentration was imaged and analyzed. Finally, for efferocytosis, the intensity of pHrodo Red dye, a pH-sensitive dye that increases in intensity with acidic pH, as found in the endolysosomes, was measured in the pHrodo Red color channel and analyzed. Results IBR significantly inhibited ADCP at all measured drug concentrations (0.41 μM, p < 0.05; 1.2 μM, p < 0.01; 3.7 - 100 μM, p < 0.001). The mean peak free drug concentration (Cmax) achieved clinically by standard doses for IBR is ~0.5 μM. ACALA only significantly inhibited ADCP at the highest concentration (100 μM, p < 0.001). The Cmax achieved clinically by standard doses for ACALA is ~1.2 μM. ACALA did not inhibit efferocytosis or subsequent transition to endolysosomal compartment at all tested concentrations (p > 0.05). IBR did not inhibit efferocytosis (p > 0.05) and only inhibited transition to endolysosomal compartment at highest concentration tested (10 μM, p < 0.01) Conclusion Our study shows that BTK inhibition does not block ADCP and a more selective BTK inhibitor may prove effective in combination with therapeutic anti-CD20 mAbs. IBR off-target inhibition specifically blocks ADCP and not efferocytosis. Thus, IBR off-target inhibition of ADCP should be via proximal signaling by antibody Fc receptors and not subsequent downstream phagocytic mechanisms in common with efferocytosis. These results also imply the lack of BTK and IBR off-target molecules involvement in efferocytosis. Finally, the increased lymphocytosis seen with IBR compared to ACALA treatment in CLL cannot be explained by IBR off-target effects on efferocytosis. These findings provide a critical understanding of macrophage phagocytosis reduction by BTK inhibitor selectivity that will have important consequences for the development of combination targeted therapies with mAbs. Disclosures Chu: Acerta Pharma/AstraZeneca: Research Funding; Pfizer: Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months; TG Therapeutics: Research Funding. Izumi:AstraZeneca: Current equity holder in publicly-traded company; Acerta Pharma: Current equity holder in private company, Ended employment in the past 24 months, Patents & Royalties: Acalabrutinib patents (no royalties). Munugalavadla:Gilead Sciences: Current equity holder in publicly-traded company; AstraZeneca: Current equity holder in publicly-traded company; Acerta Pharma: Current Employment. Barr:Gilead: Consultancy; Morphosys: Consultancy; TG therapeutics: Consultancy, Research Funding; Seattle Genetics: Consultancy; Celgene: Consultancy; AstraZeneca: Consultancy, Research Funding; Janssen: Consultancy; Merck: Consultancy; Genentech: Consultancy; Abbvie/Pharmacyclics: Consultancy, Research Funding; Verastem: Consultancy. VanDerMeid:Acerta Pharma / AstraZeneca: Research Funding. Elliott:Acerta Pharma / AstraZeneca: Research Funding. Zent:Mentrik Biotech: Research Funding; TG Therapeutics, Inc: Research Funding; Acerta / Astra Zeneca: Research Funding.

Laboratory assays used to rank potato cultivar tolerance to blackleg showed that tuber vacuum infiltration results correlate with field observations

Seed potato certification data collected in Colorado from 2012-2016 was used to rank potato cultivar tolerance to blackleg. Five cultivars with different tolerance levels to blackleg (Chipeta ≅ Alegria ≅ Lamoka < Classic Russet < Yukon Gem) were tested to determine if we could validate field data with laboratory assays. A strain isolated from Colorado, Pectobacterium atrosepticum CW1-4, and the P. atrosepticum type strain, 33260, were used to inoculate plants through vacuum infiltration of tubers or stem inoculation. Disease was assessed with time-lapse video and by measuring lesion length and disease incidence. After vacuum infiltration of tubers with P. atrosepticum CW1-4 and 33260, cultivars varied in tolerance to soft rot (Lamoka < Classic Russet < Alegria ≅ Yukon Gem < Chipeta). Blackleg tolerance of the five cultivars also varied after vacuum infiltration (Lamoka < Alegria ≅ Chipeta < Classic Russet ≅ Yukon Gem). All cultivars were susceptible after stem inoculation with either strain. In this assay, Chipeta had the longest lesions and Lamoka had the smallest lesions. Time-lapse video was used to assess Classic Russet and Yukon Gem. Yukon Gem developed disease symptoms faster than Classic Russet, but the difference was not significant. These results indicate that relative susceptibility of the five cultivars to P. atrosepticum depends on the assay used and that laboratory and greenhouse results differed from field observations.

Fertilization and Cleavage Axes Differ In Primates Conceived By Conventional (IVF) Versus Intracytoplasmic Sperm Injection (ICSI)

With nearly ten million babies conceived globally, using assisted reproductive technologies, fundamental questions remain; e.g., How do the sperm and egg DNA unite? Does ICSI have consequences that IVF does not? Here, pronuclear and mitotic events in nonhuman primate zygotes leading to the establishment of polarity are investigated by multidimensional time-lapse video microscopy and immunocytochemistry. Multiplane videos after ICSI show atypical sperm head displacement beneath the oocyte cortex and eccentric para-tangential pronuclear alignment compared to IVF zygotes. Neither fertilization procedure generates incorporation cones. At first interphase, apposed pronuclei align obliquely to the animal-vegetal axis after ICSI, with asymmetric furrows assembling from the male pronucleus. Furrows form within 30° of the animal pole, but typically, not through the ICSI injection site. Membrane flow drives polar bodies and the ICSI site into the furrow. Mitotic spindle imaging suggests para-tangential pronuclear orientation, which initiates random spindle axes and minimal spindle:cortex interactions. Parthenogenetic pronuclei drift centripetally and assemble astral spindles lacking cortical interactions, leading to random furrows through the animal pole. Conversely, androgenotes display cortex-only pronuclear interactions mimicking ICSI. First cleavage axis determination in primates involves dynamic cortex-microtubule interactions among male pronuclei, centrosomal microtubules, and the animal pole, but not the ICSI site.

AESTHETIC VALUES ON TIME LAPSE AND CINEMATIC VIDEOS

Perkembangan teknologi maklumat semakin mendorong manusia untuk membangun dan mencipta inovasi. Teknologi dapat mengembangkan potensi manusia dalam mencipta produk moden. Transformasi teknologi yang pesat dari semasa ke semasa dengan meluasnya perkembangan media audiovisual, serta teknik dalam penggambaran video seperti selang masa dan sinematik. Kajian ini bertujuan untuk mengkaji nilai estetika teknik video selang masa dan sinematik dengan mengambil kajian kes dua video, iaitu Exploring Cultural Heritage: The Old Wall in Solo and The Beauty of the City of Solo (Surakarta). Analisis dilakukan dalam lima aspek, iaitu keunikan idea, isi atau mesej, aspek komunikatif, nilai estetika, dan kualiti teknik penggambaran dan teknik suara. Penulis memfokuskan pada analisis video teknik selang masa dan teknik video sinematik berdasarkan nilai estetik yang muncul. Nilai estetik kedua-dua video dapat dilihat dari penggunaan teknik selang masa yang mendorong perspektif penonton mengenai aktiviti semula jadi dan aktiviti sosial yang berlaku dari sesuatu objek. Setelah disatukan ke dalam projek media audio-visual, keindahan objek dapat dipaparkan. Sebaliknya, perspektif yang dihasilkan melalui teknik video sinematik memberikan rasa objek, iaitu video yang mewakili kemegahan objek warisan budaya dengan sudut tinggi terus bergerak di atas objek. Hal Ini mendorong penonton untuk memperoleh maklumat dan nilai, yang disertakan melalui teknik video selang masa dan teknik video sinematik. The development of information technology increasingly encourages people to develop and create innovation. Technology can develop human potential in creating a modern product. The rapid technological transformation from time to time is marked by the widespread of developing audiovisual media, as well as by techniques in video shooting such as time lapse and cinematic. This study examines the aesthetic values of time lapse and cinematic video techniques by taking a case study of two videos, namely Exploring Cultural Heritage: The Old Wall in Solo and The Beauty of the City of Solo (Surakarta). Analysis was done in five aspects, namely the uniqueness of ideas, content or message, communicative aspects, aesthetic values, and the quality of shooting and sound techniques. The authors focused on video analysis of time-lapse video technique and cinematic video technique based on aesthetic value which emerge. The aesthetic value of both videos can be seen from the use of time-lapse technique that encourages perspective viewers about the natural and social activities that occur from an object. Having been assembled into an audio-visual media project, the beauty of an object can be projected. On the other hand, the perspective developed through cinematic technique video media is to provide a sense of object, a video that represents the splendor of a cultural heritage object with a high angle continuously moving over the object. This is to lead the audience to be able to obtain messages and values, which are embedded through both time-lapse video technique and cinematic video technique.