Iron Fortification and Inulin Supplementation in Early Infancy: Evaluating the Impact on Iron Metabolism and Trace Mineral Status in a Piglet Model.

Background: Infant formula in the United States contains abundant iron, raising health concerns about excess iron intake in early infancy. Objectives: Using a piglet model, we explored the impact of high iron fortification and prebiotic or synbiotic supplementation on iron homeostasis and trace mineral bioavailability. Methods: Twenty-four piglets were stratified and randomly assigned to treatments on postnatal day 2. Piglets were individually housed and received an iron-adequate milk diet (AI), a high-iron milk diet (HI), HI supplemented with 5% inulin (HI with a prebiotic [HIP]), or HIP with an oral gavage of Ligilactobacillus agilis YZ050, an inulin-fermenting strain, every third day (HI with synbiotic [HIS]). Milk was provided in 14 meals daily, mimicking formula feeding in infants. Fecal consistency score and body weight were recorded daily or every other day. Blood and feces were sampled weekly, and tissues collected on postnatal day 29. Data were analyzed using mixed model analysis of variance with repeated measures whenever necessary. Results: Diet did not affect growth. HI increased hemoglobin, hematocrit, and serum iron compared to AI. Despite marginal adequacy, AI upregulated iron transporter genes and maintained satisfactory iron status in most pigs. HI upregulated hepcidin gene expression in liver, caused pronounced tissue iron deposition, and markedly increased colonic and fecal iron. Inulin supplementation, regardless of L. agilis YZ050, not only attenuated hepatic iron overload but also decreased colonic and fecal iron without altering pH or the expression of iron regulatory genes. HI lowered zinc (Zn) and copper (Cu) in the duodenum and liver compared to AI, whereas HIP and HIS further decreased Zn and Cu in the liver and diminished colonic and fecal trace minerals. Conclusions: Early-infancy excessive iron fortification causes iron overload and compromises Zn and Cu absorption. Inulin decreases trace mineral absorption likely by enhancing gut peristalsis and stool frequency.

Ultrathin silicon nitride microchip for in situ/operando microscopy with high spatial resolution and spectral visibility.

Utilization of in situ/operando methods with broad beams and localized probes has accelerated our understanding of fluid-surface interactions in recent decades. The closed-cell microchips based on silicon nitride (SiNx) are widely used as "nanoscale reactors" inside the high-vacuum electron microscopes. However, the field has been stalled by the high background scattering from encapsulation (typically ~100 nanometers) that severely limits the figures of merit for in situ performance. This adverse effect is particularly notorious for gas cell as the sealing membranes dominate the overall scattering, thereby blurring any meaningful signals and limiting the resolution. Herein, we show that by adopting the back-supporting strategy, encapsulating membrane can be reduced substantially, down to ~10 nanometers while maintaining structural resiliency. The systematic gas cell work demonstrates advantages in figures of merit for hitherto the highest spatial resolution and spectral visibility. Furthermore, this strategy can be broadly adopted into other types of microchips, thus having broader impact beyond the in situ/operando fields.

Single-Molecule Graphene Liquid Cell Electron Microscopy for Instability of Intermediate Amyloid Fibrils.

Single-molecule techniques are powerful microscopy methods that provide new insights into biological processes. Liquid-phase transmission electron microscopy (LP-TEM) is an ideal single-molecule technique for overcoming the poor spatiotemporal resolution of optical approaches. However, single-molecule LP-TEM is limited by several challenges such as electron-beam-induced molecular damage, difficulty in identifying biomolecular species, and a lack of analytical approaches for conformational dynamics. Herein, a single-molecule graphene liquid-cell TEM (GLC-TEM) technique that enables the investigation of real-time structural perturbations of intact amyloid fibrils is presented. It is demonstrated that graphene membranes significantly extend the observation period of native amyloid beta proteins without causing oxidative damage owing to electron beams, which is necessary for imaging. Stochastic and time-resolved investigations of single fibrils reveal that structural perturbations in the early fibrillar stage are responsible for the formation of various amyloid polymorphs. The advantage of observing structural behavior in real time with unprecedented resolution will potentially make GLC-TEM a complementary approach to other single-molecule techniques.

Dietary butyrate and valerate glycerides impact diarrhea severity and immune response of weaned piglets under ETEC F4-ETEC F18 coinfection conditions.

Enterotoxigenic Escherichia coli (ETEC) causes post-weaning diarrhea in piglets, significantly impacting animal welfare and production efficiency. The two primary ETEC pathotypes associated with post-weaning diarrhea are ETEC F4 and ETEC F18. During the post-weaning period, piglets may be exposed to both ETEC F4 and ETEC F18. However, the effects of coinfection by both strains have not been studied. Short chain fatty acid feed additives, such as butyrate and valerate, are being investigated for their potential to improve animal performance and disease resistance. Therefore, this pilot experiment aimed to test the effects of butyrate glycerides or valerate glycerides on growth performance, diarrhea incidence, and immune responses of piglets under ETEC F4-ETEC F18 coinfection conditions. Twenty piglets were individually housed and assigned to one of the three dietary treatments immediately at weaning (21 to 24 d of age). The dietary treatments included control (basal diet formulation), control supplemented with 0.1% butyrate glycerides or 0.1% valerate glycerides. After a 7-d adaptation, all pigs were inoculated with ETEC F4 and ETEC F18 (0.5 × 109 CFU/1.5 mL dose for each strain) on three consecutive days. Pigs and feeders were weighed throughout the trial to measure growth performance. Fecal cultures were monitored for hemolytic coliforms, and blood samples were collected for whole blood and serum analysis. Pigs fed valerate glycerides tended (P = 0.095) to have higher final body weight compared with control. The overall severity of diarrhea was significantly (P < 0.05) lower in both treatment groups than control. Pigs fed valerate glycerides tended (P = 0.061) to have lower neutrophils and had significantly (P < 0.05) lower serum TNF-α on day 4 post-inoculation. This pilot experiment established an appropriate experimental dose for an ETEC F4-ETEC F18 coinfection disease model in weaned piglets. Results also suggest that butyrate glycerides and valerate glycerides alleviated diarrhea and regulated immune responses in piglets coinfected with ETEC F4 and ETEC F18.

Piglets suffer from post-weaning diarrhea associated with Enterotoxigenic Escherichia coli (ETEC) F4 and F18, two prevalent strains on swine farms globally. Short chain fatty acids (SCFAs), such as butyrate and valerate, are natural, organic compounds that could potentially promote intestinal health when used as dietary supplements. During the post-weaning period, piglets are vulnerable to simultaneous infection by ETEC F4 and F18. Therefore, this experiment aimed to develop an experimental disease model for coinfection with ETEC F4 and F18, employing a dose of 0.5 × 109 CFU/1.5 mL of each strain, administered over three consecutive days. In addition, the experiment evaluated treatment diets supplemented with 0.1% butyrate or valerate glycerides compared with the control diet. Results from this experiment revealed that the inoculation dose incited infection and diarrhea in piglets, implying its suitability for use in a disease challenge model. Moreover, the results indicated that the inclusion of butyrate and valerate glycerides to pig's diet reduced the severity of diarrhea. Furthermore, pigs fed SCFA glycerides exhibited lowered levels of inflammatory blood markers. In conclusion, the experimental dose induced diarrhea in piglets, and dietary supplementation of butyrate and valerate glycerides alleviated the severity of diarrhea while augmenting inflammatory status.

Mid-to-late Holocene climate variability in coastal East Asia and its impact on ancient Korean societies.

The sustainability of human societies is contingent upon our ability to accurately predict the effects of future climate change on the global environment and humanity. Wise responses to forthcoming environmental alterations require extensive knowledge from historical precedents. However, in coastal East Asia, a region with a long history of agriculture, it is challenging to obtain paleoenvironmental proxy data without anthropogenic disturbances that can be used to assess the impact of late Holocene climate change on local communities. This study introduces a high-resolution multi-proxy sedimentary record from an isolated crater in Jeju Island, Korea, to elucidate the mechanisms underlying mid-to-late Holocene climate change and its impacts on ancient societies. Our findings suggest that hydroclimate changes were predominantly governed by sea surface temperature fluctuations in the western tropical Pacific, with low-frequency variability in solar activity and a decrease in summer insolation identified as primary drivers of temperature change. Moreover, ancient societies on the Korean peninsula were significantly affected by recurring cooling events, including the 2.8 ka event, 2.3 ka event, Late Antique Little Ice Age, maunder minimum, and others.

Full C-band wavelength-tunable, 250 MHz repetition rate mode-locked polarization-maintaining fiber laser.

We demonstrate a full C-band wavelength-tunable mode-locked fiber laser with a repetition rate of 250 MHz, representing the highest repetition rate for C-band tunable mode-locked lasers thus far to the best of our knowledge. The polarization-maintaining fiber-based Fabry-Perot cavity enables a fundamental repetition rate of 250 MHz with a semiconductor saturable absorber mirror as a mode-locker. We observed a stable and single soliton mode-locking state with wide tunability of the center wavelength from 1505 to 1561 nm by adjusting the incident angle of a bandpass filter inside the cavity. The wavelength-tunable high-repetition-rate mode-locked laser covering the full C-band is expected to be a compelling source for many frequency-comb-based applications, including high-precision optical metrology, broadband absorption spectroscopy, and broadband optical frequency synthesizers.

Iron Deficiency and Overload Modulate the Inflammatory Responses and Metabolism of Alveolar Macrophages.

Alveolar macrophages (AM) are critical to defense against respiratory pathogens. This study evaluated cellular iron imbalance to immunometabolism in endotoxin-polarized porcine AMs (PAMs). PAMs collected from five 6-week-old pigs were treated with a basal media, iron chelator, or ferric ammonium citrate to maintain iron replete or induce iron deficiency or overload, respectively. After 24 h treatment, PAMs were challenged with saline or lipopolysaccharide (LPS) for 6 h. Cells were analyzed for gene, protein, and untargeted metabolome. Cytokines were determined in culture media. Data were assessed using two-way ANOVA. Treatments successfully induced iron deficiency and overload. The mRNA of DMT1 and ZIP14 was increased up to 300-fold by LPS, but unaffected by iron. Surprisingly, both iron deprivation and overload attenuated LPS-induced inflammation, showing less TNFα production and lower mRNA of pro- and anti-inflammatory cytokines than iron-replete PAMs. Forty-eight metabolites were altered by either or both main effects. LPS enhanced the glycolysis and polyol pathways. Iron deprivation disrupted the TCA cycle. Iron overload increased intracellular cholesterol. Interestingly, iron deprivation augmented, whereas iron overload diminished, LPS-induced itaconic acid production, which has anti-microbial and anti-inflammatory properties. Therefore, iron-deficient PAMs may be more resistant to intracellular pathogens which use PAMs as a conduit for infection.

Foot contact forces can be used to personalize a wearable robot during human walking.

Individuals with below-knee amputation (BKA) experience increased physical effort when walking, and the use of a robotic ankle-foot prosthesis (AFP) can reduce such effort. The walking effort could be further reduced if the robot is personalized to the wearer using human-in-the-loop (HIL) optimization of wearable robot parameters. The conventional physiological measurement, however, requires a long estimation time, hampering real-time optimization due to the limited experimental time budget. This study hypothesized that a function of foot contact force, the symmetric foot force-time integral (FFTI), could be used as a cost function for HIL optimization to rapidly estimate the physical effort of walking. We found that the new cost function presents a reasonable correlation with measured metabolic cost. When we employed the new cost function in HIL ankle-foot prosthesis stiffness parameter optimization, 8 individuals with simulated amputation reduced their metabolic cost of walking, greater than 15% (p < 0.02), compared to the weight-based and control-off conditions. The symmetry cost using the FFTI percentage was lower for the optimal condition, compared to all other conditions (p < 0.05). This study suggests that foot force-time integral symmetry using foot pressure sensors can be used as a cost function when optimizing a wearable robot parameter.

Stability of Plant Leaf-Derived Extracellular Vesicles According to Preservative and Storage Temperature.

Plant-derived extracellular vesicles (EVs) are capable of efficiency delivering mRNAs, miRNAs, bioactive lipids, and proteins to mammalian cells. Plant-derived EVs critically contribute to the ability of plants to defend against pathogen attacks at the plant cell surface. They also represent a novel candidate natural substance that shows potential to be developed for food, cosmetic, and pharmaceutical products. However, although plant-derived EVs are acknowledged as having potential for various industrial applications, little is known about how their stability is affected by storage conditions. In this study, we evaluated the stability of Dendropanax morbifera leaf-derived extracellular vesicles (LEVs) alone or combined with the preservatives, 1,3-butylene glycol (to yield LEVs-1,3-BG) or TMO (LEVs-TMO). We stored these formulations at -20, 4, 25, and 45 °C for up to 4 weeks, and compared the stability of fresh and stored LEVs. We also assessed the effect of freeze-thawing cycles on the quantity and morphology of the LEVs. We found that different storage temperatures and number of freeze-thawing cycles altered the stability, size distribution, protein content, surface charge, and cellular uptake of LEVs compared to those of freshly isolated LEVs. LEVs-TMO showed higher stability when stored at 4 °C, compared to LEVs and LEVs-1,3-BG. Our study provides comprehensive information on how storage conditions affect LEVs and suggests that the potential industrial applications of plant-derived EVs may be broadened by the use of preservatives.

Facile fabrication of flexible metal grid transparent electrode using inkjet-printed dot array as sacrificial layer.

In this study, we introduce a flexible metal grid transparent electrode fabricated using a lift-off process. This transparent electrode consisting of metal thin film with punched-like pattern by hole array was fabricated with 8 um separations. The separation of inkjet-printed etching resistant ink droplets was controlled in order to investigate the relationship between its electrical and optical properties of the electrodes. The aluminum areal density was defined to predict the electrical and optical properties of different arrays. A high and uniform transmittance spectrum appears to extend broadly into the UV region. The figure of merit of the transparent electrode was investigated in order to determine its performance as a transparent electrode. Moreover, there was no significant change in the resistance after 7000 bending cycles, indicating that the array conductor had superior stability. We also demonstrate transparent touch screen panels fabricated using the transparent electrode.

Linear-cavity Er-doped fiber mode-locked laser with large wavelength tunability.

A linear-type wavelength-tunable all-polarization-maintaining fiber mode-locked laser is proposed for the first time, to our knowledge, and is implemented with an Er-doped fiber and polarization-maintaining fiber components. The tuning range of the center wavelength is from 1533.7 nm to 1565.6 nm. The linear-type configuration makes the proposed laser simpler and more compact, allowing it to achieve the highest repetition rate of 126.5 MHz among C-band wavelength-tunable mode-locked lasers due to its short cavity length. Also, its polarization-maintaining fiber components provide reliable operating robustness. The significant wavelength tunability and high repetition rate of the proposed laser can be expected to make it an attractive resource for various applications, including optical communications, broadband spectroscopic LIDAR, and high-precision ranging.

Comparative physiological and transcriptomic analysis reveals salinity tolerance mechanisms in Sorghum bicolor (L.) Moench.

MAIN CONCLUSION: Mota Maradi is a sorghum line that exhibits holistic salinity tolerance mechanisms, making it a viable potential donor in breeding efforts for improved sorghum lines. High soil salinity is one of the global challenges for crop growth and productivity. Understanding the salinity tolerance mechanisms in crops is necessary for genetic breeding of salinity-tolerant crops. In this study, physiological and molecular mechanisms in sorghum were identified through a comparative analysis between a Nigerien salinity-tolerant sorghum landrace, Mota Maradi, and the reference sorghum line, BTx623. Significant differences on physiological performances were observed, particularly on growth and biomass gain, photosynthetic rate, and the accumulation of Na+, K+, proline, and sucrose. Transcriptome profiling of the leaves, leaf sheaths, stems, and roots revealed contrasting differentially expressed genes (DEGs) in Mota Maradi and BTx623 which supports the physiological observations from both lines. Among the DEGs, ion transporters such as HKT, NHX, AKT, HAK5, and KUP3 were likely responsible for the differences in Na+ and K+ accumulation. Meanwhile, DEGs involved in photosynthesis, cellular growth, signaling, and ROS scavenging were also identified between these two genotypes. Functional and pathway analysis of the DEGs has revealed that these processes work in concert and are crucial in elevated salinity tolerance in Mota Maradi. Our findings indicate how different complex processes work synergistically for salinity stress tolerance in sorghum. This study also highlights the unique adaptation of landraces toward their respective ecosystems, and their strong potential as genetic resources for future plant breeding endeavors.

Optical method for simultaneous thickness measurements of two layers with a significant thickness difference.

In this study, an optical method that allows simultaneous thickness measurements of two different layers distributed over a broad thickness range from several tens of nanometers to a few millimeters based on the integration of a spectroscopic reflectometer and a spectral-domain interferometer is proposed. Regarding the optical configuration of the integrated system, various factors, such as the operating spectral band, the measurement beam paths, and the illumination beam type, were considered to match the measurement positions and effectively separate two measurement signals acquired using both measurement techniques. Furthermore, for the thickness measurement algorithm, a model-based analysis method for high-precision substrate thickness measurements in thin-film specimens was designed to minimize the measurement error caused by thin films, and it was confirmed that the error is decreased significantly to less than 8 nm as compared to that when using a Fourier-transform analysis. The ability to undertake simultaneous thickness measurements of both layers using the proposed system was successfully verified on a specimen consisting of silicon dioxide thin film with nominal thicknesses of 100 nm and 150 nm and a 450 µm-thick silicon substrate, resulting in the exact separation between the two layers. From measurement uncertainty evaluation of a thin-film, a substrate in a thin-film specimen, and a single substrate, the uncertainties were estimated to be 0.12 nm for the thin-film, 0.094 µm for the substrate in a thin-film specimen, and 0.076 µm for the substrate. The measurement performance of thicknesses distributed on multi-scale was verified through comparative measurements using standard measurement equipment for several reference samples.

Sub-100-nm precision distance measurement by means of all-fiber photonic microwave mixing.

The importance of dimensional metrology has gradually emerged from fundamental research to high-technology industries. In the era of the fourth industrial revolution, absolute distance measurements are required to cope with various applications, such as unmanned vehicles, intelligent robots, and positioning sensors for smart factories. In such cases, the size, weight, power, and cost (SWaP-C) should essentially be restricted. In this paper, sub-100 nm precision distance measurements based on an amplitude-modulated continuous-wave laser (AMCW) with an all-fiber photonic microwave mixing technique is proposed and realized potentially to satisfy SWaP-C requirements. Target distances of 0.879 m and 8.198 m were measured by detecting the phase delay of 15 GHz modulation frequencies. According to our measurement results, the repeatability could reach 43 nm at an average time of 1 s, a result not previously achieved by conventional AMCW laser distance measurement methods. Moreover, the performance by the proposed method in terms of Allan deviation is competitive with most frequency-comb-based absolute distance measurement methods, even with a simple configuration. Because the proposed method has a simple configuration such that it can be easily utilized and demonstrated on a chip-scale platform using CMOS-compatible silicon photonics, it is expected to herald new possibilities, leading to the practical realization of a fully integrated chip-scale LIDAR system.

Cyclic tangential flow filtration system for isolation of extracellular vesicles.

Size-based filtration techniques have been developed for high-throughput isolation of extracellular vesicles (EVs). Conventional direct filtration systems have limitations in that large particles generally not only block the pores of the membrane but also damage the particles because of the high fluid pressure. Here, we propose a cyclic tangential flow filtration (TFF) system that includes two membranes with pore sizes of 200 and 30 nm, connected to a peristaltic pump that feeds the stream flowing to the membrane for continuous circulation. The cyclic TFF system is better able to isolate the specific 30-200 nm size range in one step through dual cyclic filtration compared with direct filtration (DF) and single cyclic TFF (scTFF). We further introduced a buffer-exchange process to the dcTFF system after filtration to remove contaminants for more efficient purification. As a result of comparative evaluation of dcTFF and ExoQuick, EVs isolated by dcTFF had more abundant exosome markers and active EVs. The cyclic TFF system not only has great potential to separate EVs with high selectivity and separation efficiency in small volumes of samples but can also be used in clinical applications, including medical diagnostic procedures.

Graphene Liquid Cell Electron Microscopy: Progress, Applications, and Perspectives.

Graphene liquid cell electron microscopy (GLC-EM), a cutting-edge liquid-phase EM technique, has become a powerful tool to directly visualize wet biological samples and the microstructural dynamics of nanomaterials in liquids. GLC uses graphene sheets with a one carbon atom thickness as a viewing window and a liquid container. As a result, GLC facilitates atomic-scale observation while sustaining intact liquids inside an ultra-high-vacuum transmission electron microscopy chamber. Using GLC-EM, diverse scientific results have been recently reported in the material, colloidal, environmental, and life science fields. Here, the developments of GLC fabrications, such as first-generation veil-type cells, second-generation well-type cells, and third-generation liquid-flowing cells, are summarized. Moreover, recent GLC-EM studies on colloidal nanoparticles, battery electrodes, mineralization, and wet biological samples are also highlighted. Finally, the considerations and future opportunities associated with GLC-EM are discussed to offer broad understanding and insight on atomic-resolution imaging in liquid-state dynamics.

Liquid-Flowing Graphene Chip-Based High-Resolution Electron Microscopy.

The recent advances in liquid-phase transmission electron microscopy represent tremendous potential in many different fields and exciting new opportunities. However, achieving both high-resolution imaging and operando capabilities remain a significant challenge. This work suggests a novel in situ imaging platform of liquid-flowing graphene chip TEM (LFGC-TEM) equipped with graphene viewing windows and a liquid exchange system. The LFGCs are robust under high-pressure gradients and rapid liquid circulation in ranges covering the experimental conditions accessible with conventional thick SiNx chips. LFGC-TEM provides atomic resolution for colloidal nanoparticles and molecular-level information limits for unstained wet biomolecules and cells that are comparable to the resolutions achievable with solid-phase and cryogenic TEM, respectively. This imaging platform can provide an opportunity for live imaging of biological phenomena that is not yet achieved using any current methods.

Lithographically patterned well-type graphene liquid cells with rational designs.

Graphene liquid cell transmission electron microscopy allows in situ observation of nanomaterial dynamics in a liquid environment. However, this method suffers from both random formation and small size of liquid pockets. Here, we introduce facile and mass-producible graphene-sealed well-type liquid cells with rational designs. The developed liquid cell structure and its formation mechanism depending on hole diameter (d)/spacer thickness (h) ratio are systematically analyzed. Finally, we show its high-resolution imaging and chemical analysis capability for nanoparticles and biomaterial applications. This work will provide an enhanced liquid cell platform for diverse liquid environmental studies.

Precise thickness profile measurement insensitive to spatial and temporal temperature gradients on a large glass substrate.

When manufacturing glass substrates for display devices, especially for large-sized ones, the time-varying spatial temperature gradient or distribution on the samples is remarkably observed. It causes serious degradation of thickness measurement accuracy due to the combination of thermally expanded thickness and temperature-dependent refractive index. To prevent or minimize the degradation in thickness measurement accuracy, the temperature distribution over an entire glass substrate has to be known in real time in synchronization with the thickness measurement to specify the refractive index of the sample based on an exact mathematical model of the temperature-dependent refractive index. In this paper, a measurement method for determining the thickness profile of a large glass substrate regardless of precise measurement of temperature distribution and the mathematical model of the refractive index was demonstrated. The widely used glass substrates with nominal thicknesses of 0.6 mm and 1.3 mm were measured at room and high temperatures. Through comparison of thickness profiles of hot glass substrates having large temperature gradients and those estimated through thermal expansion of thickness profiles measured at room temperature, it was confirmed that the proposed method can provide highly reliable thickness measurement results under such challenging conditions, unlike simple calculation from the optical thickness using the well-known refractive index.

Real-Time Observation of CaCO3 Mineralization in Highly Supersaturated Graphene Liquid Cells.

The mineralization dynamics of calcium carbonate is investigated under highly supersaturated conditions using graphene liquid cell transmission electron microscopy. We demonstrate that the mineralization process has three steps: nucleation, diffusion-limited growth, and Ostwald ripening/coalescence. In addition, we show that the polymorphs precipitate in a specific order, from metastable aragonite to stable calcite, thus proving Ostwald's rule of stages. In highly supersaturated solutions, the aragonite phase crystallizes in a stable manner, in addition to the calcite phase.