Extended culture of 2D gastruloids to model human mesoderm development.

Micropatterned human pluripotent stem cells (hPSCs) treated with BMP4 (2D gastruloids) are among the most widely used stem cell models for human gastrulation. Due to its simplicity and reproducibility, this system is ideal for high throughput quantitative studies of tissue patterning and has led to many insights into the mechanisms of mammalian gastrulation. However, 2D gastruloids have only been studied up to 48h. Here we extended this system to 96h. We discovered a phase of highly reproducible morphogenesis during which directed migration from the primitive streak-like region gives rise to a mesodermal layer beneath an epiblast-like layer. Multiple types of mesoderm arise with striking spatial organization including lateral mesoderm-like cells on the colony border and paraxial mesoderm-like further inside the colony. Single cell transcriptomics showed strong similarity of these cells to mesoderm in human and non-human primate embryos. However, our data suggest that the annotation of the reference human embryo may need to be revised. This illustrates that extended culture of 2D gastruloids provides a powerful model for human mesoderm differentiation and morphogenesis.

Time-integrated BMP signaling determines fate in a stem cell model for early human development.

How paracrine signals are interpreted to yield multiple cell fate decisions in a dynamic context during human development in vivo and in vitro remains poorly understood. Here we report an automated tracking method to follow signaling histories linked to cell fate in large numbers of human pluripotent stem cells (hPSCs). Using an unbiased statistical approach, we discover that measured BMP signaling history correlates strongly with fate in individual cells. We find that BMP response in hPSCs varies more strongly in the duration of signaling than the level. However, both the level and duration of signaling activity control cell fate choices only by changing the time integral. Therefore, signaling duration and level are interchangeable in this context. In a stem cell model for patterning of the human embryo, we show that signaling histories predict the fate pattern and that the integral model correctly predicts changes in cell fate domains when signaling is perturbed. Our data suggest that mechanistically, BMP signaling is integrated by SOX2.

Development of a Nurse Turnover Prediction Model in Korea Using Machine Learning.

Nurse turnover is a critical issue in Korea, as it affects the quality of patient care and increases the financial burden on healthcare systems. To address this problem, this study aimed to develop and evaluate a machine learning-based prediction model for nurse turnover in Korea and analyze factors influencing nurse turnover. The study was conducted in two phases: building the prediction model and evaluating its performance. Three models, namely, decision tree, logistic regression, and random forest were evaluated and compared to build the nurse turnover prediction model. The importance of turnover decision factors was also analyzed. The random forest model showed the highest accuracy of 0.97. The accuracy of turnover prediction within one year was improved to 98.9% with the optimized random forest. Salary was the most important decision factor for nurse turnover. The nurse turnover prediction model developed in this study can efficiently predict nurse turnover in Korea with minimal personnel and cost through machine learning. The model can effectively manage nurse turnover in a cost-effective manner if utilized in hospitals or nursing units.

The time integral of BMP signaling determines fate in a stem cell model for early human development.

How paracrine signals are interpreted to yield multiple cell fate decisions in a dynamic context during human development in vivo and in vitro remains poorly understood. Here we report an automated tracking method to follow signaling histories linked to cell fate in large numbers of human pluripotent stem cells (hPSCs). Using an unbiased statistical approach, we discovered that measured BMP signaling history correlates strongly with fate in individual cells. We found that BMP response in hPSCs varies more strongly in the duration of signaling than the level. However, we discovered that both the level and duration of signaling activity control cell fate choices only by changing the time integral of signaling and that duration and level are therefore interchangeable in this context. In a stem cell model for patterning of the human embryo, we showed that signaling histories predict the fate pattern and that the integral model correctly predicts changes in cell fate domains when signaling is perturbed. Using an RNA-seq screen we then found that mechanistically, BMP signaling is integrated by SOX2.

Stability and Antibiofilm Efficiency of Slightly Acidic Electrolyzed Water Against Mixed-Species of Listeria monocytogenes and Staphylococcus aureus.

In the natural environment, most microorganisms live in mixed-species biofilms, in which the metabolism and growth of organisms are different from that in single-species biofilms. Adhesive bacteria and their biofilms on the surface of food processing equipment are the sources of cross-contamination, leading to the risk for humans. Slightly acidic electrolyzed water (SAEW) has been proposed as a novel sanitizer in the food and agriculture industry. In this study, we investigated the changes in the physical properties of SAEW under different conditions and the disinfection abilities of SAEW against spore-forming and non-spore-forming pathogens. Furthermore, we examined the disinfection abilities of SAEW after 12 months of shelf life on a mixed-species biofilm of Listeria monocytogenes Scott A and Staphylococcus aureus. The results showed that SAEW at 30 and 50 ppm achieved all-kill of the spore-forming pathogen Bacillus cereus within 30 s. Changes in the ACC and pH of the produced SAEW were generally affected by the storage conditions. Both spore-forming and non-spore-forming pathogens were not detected under treatment with 50 ppm SAEW for 5 min under HDPE-closed conditions throughout the whole storage period. Moreover, 25 mg/L SAEW can inactivate L. monocytogenes Scott A and S. aureus biofilm cells in ~2.45 and 2.57 log CFU/mL in biofilms within 5-min treatment. However, the decline of the two bacteria in the mixed-species biofilm was 1.95 and 1.43 log CFU/mL, respectively. The changes in the cell membrane permeability of the mixed-species biofilm under treatment with SAEW were observed by using atomic force microscopy and confocal laser scanning microscopy. L. monocytogenes Scott A was more sensitive to SAEW in the mixed-species biofilm cells. These findings exhibited strong antibiofilm activities of SAEW in impairing biofilm cell membranes, decreasing cell density, and eliminating biofilm, which suggest that SAEW is an excellent antibacterial agent in the food processing industries.

Efficient differentiation of human primordial germ cells through geometric control reveals a key role for Nodal signaling.

Human primordial germ cells (hPGCs) form around the time of implantation and are the precursors of eggs and sperm. Many aspects of hPGC specification remain poorly understood because of the inaccessibility of the early postimplantation human embryo for study. Here, we show that micropatterned human pluripotent stem cells (hPSCs) treated with BMP4 give rise to hPGC-like cells (hPGCLC) and use these as a quantitatively reproducible and simple in vitro model to interrogate this important developmental event. We characterize micropatterned hPSCs up to 96 hr and show that hPGCLC populations are stable and continue to mature. By perturbing signaling during hPGCLC differentiation, we identify a previously unappreciated role for Nodal signaling and find that the relative timing and duration of BMP and Nodal signaling are critical parameters controlling the number of hPGCLCs. We formulate a mathematical model for a network of cross-repressive fates driven by Nodal and BMP signaling, which predicts the measured fate patterns after signaling perturbations. Finally, we show that hPSC colony size dictates the efficiency of hPGCLC specification, which led us to dramatically improve the efficiency of hPGCLC differentiation.

In humans and other animals, eggs and sperm are unique cells that pass on genetic material to the next generation. They originate from a small group of cells called primordial germ cells that form early in life in the developing embryo. Several different signal molecules including ones known as BMP4, Wnt, and Nodal, instruct certain cells in the embryo to become primordial germ cells. The process by which primordial germ cells are made in humans is very different to how primordial germ cells are made in mice and other so-called model animals that are commonly used in research. This has made it more challenging to uncover the details of the process in humans. Fortunately, new methods have recently been created that mimic aspects of how human embryos develop using human stem cells in a laboratory dish, providing an opportunity to gain a deeper understanding of how human germ cells form. Jo et al. used a technique called micropatterning to control the shape and size of groups of human stem cells growing in a laboratory dish. Treating these cells with a signal known as BMP4 gave rise to cells that resembled primordial germ cells. The team then used this system as a model to study how primordial germ cells form in humans. The experiments found that reducing Wnt signals in stem cells stopped primordial germ cells from forming in response to BMP4, confirming that Wnt signals made by the cells in response to BMP4 are essential. However, this block was overcome by providing the stem cells with another signal called Nodal. This suggests that the role of Wnt signaling in primordial germ cell formation is in part indirect by switching on Nodal in stem cells. Defects in eggs and sperm may lead to infertility, therefore, the findings of Jo et al. have the potential to help researchers develop new fertility treatments that use eggs or sperm grown in a laboratory from the patients' own stem cells. Such research would benefit from first developing a better understanding of how to make primordial germ cells.

Optimization and Effect of Water Hardness for the Production of Slightly Acidic Electrolyzed Water on Sanitization Efficacy.

Slightly acidic electrolyzed water (SAEW) has been recently proposed as a novel promising sanitizer and cleaner in the agricultural and food industries. However, several factors, including water hardness, were considered to strongly affect the physical properties and sanitization efficacy of SAEW. To study the effect of water hardness on the SAEW production, we evaluated the production properties and sanitization effect of SAEW, which was generated from water sources in 16 representatively geographical locations of South Korea. The results showed that the hardness of water sources from Kangwon-do, Jeollanam-do, and Daegu was 22-41 ppm; that from Busan, Gyeongnam-do, Gwangju Bukgu was 80-443 ppm, and that from seven other locations was 41-79 ppm. SAEW is produced from water hardness less than 50 ppm and greater than 80 ppm was beyond the accepted pH range (5.0-6.5). Notably, high-hardness water (>80 ppm) containing 5% HCl could be used to produce SAEW with accepted pH. The SAEW generated from low-hardness water with additions of 2% HCl and 2 M NaCl at 7 A showed accepted pH and higher germicidal effect. Furthermore, SAEW with the available chlorine concentration of 27-41 mg/L for 1 min was sufficient to completely inactivate non-spore-forming foodborne pathogens. Sanitization efficacy was not markedly affected by storage conditions for SAEW at 40 ppm. Our results demonstrated that the degree of water hardness is an important factor in the production of SAEW, which would provide a foundation for commercial application of SAEW.

Preparation of Polydimethylsiloxane-Modified Waterborne Polyurethane Coatings for Marine Applications.

A series of waterborne polyurethane dispersions (WPUs) modified with hydroxyl-terminated polydimethylsiloxane (PDMS) were prepared by incorporating PDMS into the soft segments of polyurethane chains. The structural characteristics of the prepared samples were analyzed by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and particle size analysis (PSA). The effect of PDMS content on the thermal and mechanical properties of PDMS-modified waterborne polyurethanes (PS-WPU) was investigated. In addition, the water resistance and dimensional stability of the PS-WPU were investigated by measuring its water absorption ratio and water contact angle along with universal testing machine measurements.

Development of Nanosensors Based Intelligent Packaging Systems: Food Quality and Medicine.

The issue of medication noncompliance has resulted in major risks to public safety and financial loss. The new omnipresent medicine enabled by the Internet of things offers fascinating new possibilities. Additionally, an in-home healthcare station (IHHS), it is necessary to meet the rapidly increasing need for routine nursing and on-site diagnosis and prognosis. This article proposes a universal and preventive strategy to drug management based on intelligent and interactive packaging (I2Pack) and IMedBox. The controlled delamination material (CDM) seals and regulates wireless technologies in novel medicine packaging. As such, wearable biomedical sensors may capture a variety of crucial parameters via wireless communication. On-site treatment and prediction of these critical factors are made possible by high-performance architecture. The user interface is also highlighted to make surgery easier for the elderly, disabled, and patients. Land testing incorporates and validates an approach for prototyping I2Pack and iMedBox. Additionally, sustainability, increased product safety, and quality standards are crucial throughout the life sciences. To achieve these standards, intelligent packaging is also used in the food and pharmaceutical industries. These technologies will continuously monitor the quality of a product and communicate with the user. Data carriers, indications, and sensors are the three most important groups. They are not widely used at the moment, although their potential is well understood. Intelligent packaging should be used in these sectors and the functionality of the systems and the values presented in this analysis.

Janus Graphene Oxide Sheets with Fe3O4 Nanoparticles and Polydopamine as Anodes for Lithium-Ion Batteries.

In this study, a one-step process to fabricate "Janus"-structured nanocomposites with iron oxide (Fe3O4) nanoparticles (Fe3O4 NPs) and polydopamine (PDA) on each side of a graphene oxide (GO) nanosheet using the Langmuir-Schaefer technique has been proposed. The Fe3O4 NPs-GO hybrid is used as a high-capacity active material, while PDA is added as a binder due to its unique wet-resistant adhesive property. The transmission electron microscopy image shows a superlattice-like out-of-plane section of the multilayered nanocomposite, which maximizes the density of the composite materials. Grazing-incidence small-angle X-ray scattering results combined with scanning electron microscopy images confirm that the multilayered Janus composite exhibits an in-plane hexagonal array structure of closely packed Fe3O4 NPs. This Janus multilayered structure is expected to maximize the amount of active material in a specific volume and reduce volume changes caused by the conversion reaction of Fe3O4 NPs. According to the electrochemical results, the Janus multilayer electrode delivers an excellent capacity of ∼903 mAh g-1 at a current density of 200 mA g-1 and a reversible capacity of ∼639 mAh g-1 at 1 A g-1 up to the 1800th cycle, indicating that this Janus composite can be a promising anode for Li-ion batteries.

Asynchronous multi-decadal time-scale series of biotic and abiotic responses to precipitation during the last 1300 years.

East Asian summer monsoon (EASM)-driven rapid hydroclimatic variation is a crucial factor with major socioeconomic impacts. Nevertheless, decadal- to centennial-scale EASM variability over the last two millennia is still poorly understood. Pollen-based quantitative annual precipitation (PqPann) and annual precipitation reconstructed by artificial neural networks (ANNs) for the period 650-1940 CE were reconstructed from a paleo-reservoir in South Korea. ANNs reconstruction was performed to compensate for a hiatus section. On a decadal timescale, 10 high-precipitation periods were identified, and PqPann and ANNs reconstructions were comparable to local instrumental rainfall and historic drought records. Biotic lags to rapid climatic changes ranging from 25 to 100 years were recognized by asynchronous pollen and speleothem responses to precipitation. We suggest that PqPann-based decadal- to centennial-scale climatic change reconstruction should take biotic lags into account, although the lags can be ignored on the millennial scale. The position of the EASM rainband influenced rainfall magnitude.

Opposite response modes of NADW dynamics to obliquity forcing during the late Paleogene.

Although the responses of North Atlantic Deep Water (NADW) is deeply connected to orbital rhythms, those under different tectonic and atmospheric boundary conditions remain unknown. Here, we report suborbitally resolved benthic foraminiferal stable isotope data from J-anomaly Ridge in the North Atlantic from ca. 26.4-26.0 Ma. Our results indicate that the formation of NADW during that time interval was increased during the obliquity-paced interglacial periods, similar to in the Plio-Pleistocene. During the late Oligocene, the interglacial poleward shifts of the stronger westerlies in the southern hemisphere, which occurred due to the higher thermal contrasts near the upper limit of the troposphere, reinforced the Antarctic Circumpolar Current (ACC) and, in turn, the Atlantic meridional overturning circulation (AMOC). However, such a response mode in deep ocean circulation did not occur during the middle Eocene because of different tectonic boundary conditions and the immature states of the ACC. Instead, the middle Eocene interglacial conditions weakened the formation of the proto-type NADW due to less heat loss rate in high-latitude regions of the North Atlantic during high obliquity periods. Our findings highlight the different responses of deep ocean circulation to orbital forcing and show that climate feedbacks can be largely sensitive to boundary conditions.

Enhanced Dynamics of Confined Polymers near the Immiscible Polymer-Polymer Interface: Neutron Reflectivity Studies.

For polymer-blend films, local dynamics in confined polymer domains tend to differ from the bulk because of significant contributions from the polymer-polymer interface. Herein, we investigated the diffusion dynamics of entangled polymer thin films confined between different polymers in a direction perpendicular to the surface using neutron reflectivity. We found that a bilayer of poly(methyl methacrylate) (PMMA) and deuterated PMMA (dPMMA) sandwiched between polystyrene (PS) layers exhibited significant increase in mobility near the polymer-polymer interface with decreasing PMMA thickness. This indicates that the contribution of repulsive interactions at the immiscible polymer-polymer interface becomes more significant as the film thickness decreases. We also found that the interfacial roughness between PMMA and PS (28 Å at equilibrium) and soft confinement of PS layers did not significantly affect the change in the diffusion dynamics of the adjacent PMMA. This was evidenced by comparison with the diffusion results of multilayers with a flat interface (8 Å at equilibrium) between PMMA and hard PS by UV cross-linking.

Position-Dependent Diffusion Dynamics of Entangled Polymer Melts Nanoconfined by Parallel Immiscible Polymer Films.

The morphological structure and dynamics of confined polymers adjacent to the polymer-polymer interface have a profound effect on determining the overall physical properties of polymer blends. We measured the diffusion dynamics of poly(methyl methacrylate) (PMMA) melts confined between polystyrene (PS) layers using neutron reflectivity. Combinations of various thicknesses of PMMA and deuterated PMMA (dPMMA) allowed us to experimentally reveal the nonmonotonic behavior of polymer mobility near the PS-PMMA interface. From the neutron reflectivity results, we found that the polymers adjacent to the immiscible polymer-polymer interface showed enhanced diffusion dynamics because of the repulsive interaction between PS and PMMA, whereas the polymer at local regions farther from the interface exhibited reduced dynamics. This is probably due to the nonspherical conformation of PMMA and spatial confinement near the PS-PMMA interface.

Effects of Al-Si Coating and Zn Coating on the Hydrogen Uptake and Embrittlement of Ultra-High Strength Press-Hardened Steel.

Press-hardened steel (PHS), used for automotive safety-related structure parts, is sensitive to hydrogen embrittlement due to its martensitic microstructure. Hydrogen is introduced in PHS during the hot press forming (HPF) process, by an atmospheric corrosion process. In this study, the hydrogen embrittlement behavior of uncoated, aluminized, and galvanized PHSs was investigated. The Al-10%Si coating promoted the absorption of diffusible hydrogen at elevated temperature during the HPF while the reacted coating layer prevented the absorbed hydrogen from out-diffusing through the reacted coating surface layer at room temperature. Therefore, the aluminized PHS showed a greater sensitivity to both the hydrogen uptake and the resultant embrittlement, as compared to the uncoated and galvanized PHSs. Use of galvanized PHS for HPF application reduces the risk of hydrogen embrittlement, since the Zn coating effectively prevents the hydrogen uptake. The greater embrittlement resistance of the galvanized PHS is possibly due to the inhibition of the hydrogen generation reaction by the surface ZnO oxide layer and the low rate of hydrogen transport through the liquid Zn phase.

Poc1B and Sas-6 Function Together during the Atypical Centriole Formation in Drosophila melanogaster.

Insects and mammals have atypical centrioles in their sperm. However, it is unclear how these atypical centrioles form. Drosophila melanogaster sperm has one typical centriole called the giant centriole (GC) and one atypical centriole called the proximal centriole-like structure (PCL). During early sperm development, centriole duplication factors such as Ana2 and Sas-6 are recruited to the GC base to initiate PCL formation. The centriolar protein, Poc1B, is also recruited at this initiation stage, but its precise role during PCL formation is unclear. Here, we show that Poc1B recruitment was dependent on Sas-6, that Poc1B had effects on cellular and PCL Sas-6, and that Poc1B and Sas-6 were colocalized in the PCL/centriole core. These findings suggest that Sas-6 and Poc1B interact during PCL formation. Co-overexpression of Ana2 and Sas-6 induced the formation of ectopic particles that contained endogenous Poc1 proteins and were composed of PCL-like structures. These structures were disrupted in Poc1 mutant flies, suggesting that Poc1 proteins stabilize the PCL-like structures. Lastly, Poc1B and Sas-6 co-overexpression also induced the formation of PCL-like structures, suggesting that they can function together during the formation of the PCL. Overall, our findings suggest that Poc1B and Sas-6 function together during PCL formation.

Dewetting of Thin Polymer Films on Wrinkled Graphene Oxide Monolayers.

We investigated the effect of the morphological structure of a graphene oxide (GO) monolayer on the dewetting dynamics of the upper polymer thin films. The Langmuir-Schaefer (LS) technique was used to prepare a wrinkled GO ( wrGO) structure with a root mean square (rms) roughness of 22.7 Å. The dewetting behavior of poly(methyl methacrylate) (PMMA) thin films on the wrGO monolayers was perfectly prevented, whereas the PMMA thin films on a flat GO monolayer were dewetted at 203 °C. This wrinkle effect of the GO can be also obtained when the GOs monolayers are intercalated to the PMMA/polystyrene (PS) interface. In this multilayer, the flat GO monolayer at the interface between the PS and PMMA layers was spontaneously roughened with rms roughness of 46.9 Å after annealing and also prohibited the dewetting behavior. From the results, we found that to improve the compatibility of polymer blends by adding the two-dimensional nanosheets, it is important to control the morphological structure of the sheets at the interface, along with manipulation of the GO-polymer interactions.

Author Correction: A novel atypical sperm centriole is functional during human fertilization.

In the original version of this Article, the affiliation details for Jadranka Loncarek and Vito Mennella were incorrectly given as 'Cell Biology Program, The Hospital for Sick Children, Department of Biochemistry, University of Toronto, 555 University Avenue, Toronto, ON, M5G 1X8, Canada' and 'Laboratory of Protein Dynamics and Signaling, Center for Cancer Research, National Cancer Institute, 1050 Boyles Street, Frederick, MD, 21702, USA', respectively. This has now been corrected in both the PDF and HTML versions of the Article.

A novel atypical sperm centriole is functional during human fertilization.

The inheritance of the centrosome during human fertilization remains mysterious. Here we show that the sperm centrosome contains, in addition to the known typical barrel-shaped centriole (the proximal centriole, PC), a surrounding matrix (pericentriolar material, PCM), and an atypical centriole (distal centriole, DC) composed of splayed microtubules surrounding previously undescribed rods of centriole luminal proteins. The sperm centrosome is remodeled by both reduction and enrichment of specific proteins and the formation of these rods during spermatogenesis. In vivo and in vitro investigations show that the flagellum-attached, atypical DC is capable of recruiting PCM, forming a daughter centriole, and localizing to the spindle pole during mitosis. Altogether, we show that the DC is compositionally and structurally remodeled into an atypical centriole, which functions as the zygote's second centriole. These findings now provide novel avenues for diagnostics and therapeutic strategies for male infertility, and insights into early embryo developmental defects.