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Objective Two series of experiments were conducted to determine how the incremental levels of sodium metabisulfite (SMB)-treated fruit and vegetable discards (FVD) in diet of Hanwoo heifers and cows affect their performance and health. Methods In Exp. 1, 36 Hanwoo heifers were stratified by age (13.3 ± 0.83 mo) and initial body weight (305 ± 19.7 kg), and divided randomly to one of three diets containing 0, 10, or 20% SMB-treated FVD (as-fed basis). The experiment lasted 110 d, including 20 d of adaptation. In Exp. 2, 24 multiparous Hanwoo cows were divided into three groups based on age (48.2 ± 2.81 mo) and initial body condition score (2.64 ± 0.33). Cows in each block were assigned randomly to one of three diets containing 0, 11, or 22% SMB-treated FVD (as-fed basis). The experiment lasted 80 d, including a 20-d adaptation period. In both experiments, SMB-treated FVD was used as a replacement for wet brewers grain in total mixed ration (TMR). Results Growing heifers exhibited no differences in their daily feed intake (6.58 ± 0.61 kg/d DM), average daily gain (0.60 ± 0.07 kg/d), and body condition score when they consumed the incremental levels of SMB-treated FVD. Although the majority of blood metabolites were unaffected by treatments, blood urea-N and s-hydroxybutyrate levels decreased linearly as the SMB-treated FVD level increased in TMR. Similar to Exp. 1, minor differences were found in daily feed intake (8.27 ± 0.72 kg DM/d) and body condition score of Hanwoo cows. The majority of blood metabolites remained unaffected by treatments, but blood urea-N decreased as the SMB-treated FVD level in TMR increased. Conclusion Our findings suggest that SMB-treated FVD could be safely incorporated into the diet of Hanwoo heifers and cows, potentially improving N-use efficiency in the body while not impairing performance or health.

Glycation is a spontaneous post-translational modification of lysine, arginine, and the N-terminus of proteins. Protein glycation is closely related to the pathogenesis of human diseases, including diabetes, Alzheimer’s disease, renal disease, and cancer. The levels of advanced glycation end products (AGEs) are positively correlated with the progression of many diseases. However, it remains challenging to analyze glycation-related products, such as reactive carbonyl species, Schiff bases, Amadori compounds, and AGEs, because of their high heterogeneity. Many analysis methods, such as fluorescence detection, immunoassays, and liquid chromatography-tandem mass spectrometry, have attempted to correlate glycation products with diseases. Some enrichment methods have been used to increase the probability of detection of glycated proteins due to their low abundance in blood plasma. This review summarizes the enrichment and analysis methods that are currently used to identify glycation as a disease biomarker in exploratory studies.

Aims: This study aimed to perform a quantitative analysis of the proteomic changes in neural cells under H2O2 exposure and N-acetylcysteine (NAC) treatment. Background: NAC is a potent antioxidant and an effective free radical scavenger that has been used as a potential treatment of several neurological disorders. Although the molecular mechanisms of neuroprotective action by NAC have been studied, more efforts are still needed for the elucidation of the mechanisms through a quantitative proteomic analysis. Objective: This study aimed to identify differentially expressed proteins among control, H2O2-treated group, and NAC+H2O2-treated group as well as reveal proteins involved in the protection of neural cells from H2O2-induced toxicity. Methods: SK-N-MC cells were untreated (control), treated with H2O2 (disease group), pretreated with NAC, and then treated with H2O2 (NAC group). Proteins were digested to peptides and analyzed using liquid chromatography-tandem mass spectrometry with the data-independent acquisition. Skyline was used to quantify peptides and proteins. MSstats was used for statistical analysis. Gene ontology and protein-protein interactions were performed using the Differentially Expressed Proteins (DEPs). Results: Cytoprotective effects of NAC on the cell against H2O2-induced toxicity were first proven using a cell viability study and lactate dehydrogenase assay. The proteomic analysis found 93 DEPs in three comparisons. Among them, 37 proteins were differentially expressed under H2O2 exposure. Only 10 DEPs were rescued in the case of NAC pretreatment. Aspartate aminotransferase and L-lactate dehydrogenase B chain were two DEPs involved in the cysteine and methionine metabolism pathway, which might relate to the mechanisms of NAC protective effects. Conclusion: The findings of cell studies and proteomic analysis were in agreement with previous results, confirming the cytoprotective effects of NAC on neural cells against oxidative stress.

Background: Diabetic Retinopathy (DR), one of the major microvascular complications commonly occurring in diabetic patients, can be classified into Proliferative Diabetic Retinopathy (PDR) and Non-Proliferative Diabetic Retinopathy (NPDR). Currently available therapies are only targeted for later stages of the disease in which some pathologic changes may be irreversible. Thus, there is a need to develop new treatment options for earlier stages of DR through revealing pathological mechanisms of PDR and NPDR. Objective: The purpose of this study was to characterize proteomes of diabetic through quantitative analysis of PDR and NPDR. Methods: Vitreous body was collected from three groups: control (non-diabetes mellitus), NPDR, and PDR. Vitreous proteins were digested to peptide mixtures and analyzed using LC-MS/MS. MaxQuant was used to search against the database and statistical analyses were performed using Perseus. Gene ontology analysis, related-disease identification, and protein-protein interaction were performed using the differential expressed proteins. Results: Twenty proteins were identified as critical in PDR and NPDR. The NPDR group showed different expressions of kininogen-1, serotransferrin, ribonuclease pancreatic, osteopontin, keratin type II cytoskeletal 2 epidermal, and transthyretin. Also, prothrombin, signal transducer and activator of transcription 4, hemoglobin subunit alpha, beta, and delta were particularly up-regulated proteins for PDR group. The up-regulated proteins related to complement and coagulation cascades. Statherin was down-regulated in PDR and NPDR compared with the control group. Transthyretin was the unique protein that increased its abundance in NPDR compared with the PDR and control group. Conclusion: This study confirmed the different expressions of some proteins in PDR and NPDR. Additionally, we revealed uniquely expressed proteins of PDR and NPDR, which would be differential biomarkers: prothrombin, alpha-2-HS-glycoprotein, hemoglobin subunit alpha, beta, and transthyretin.

The standard bottom-up proteomic workflow is comprised of sample preparation, data acquisition, and data analysis. While the latter two parts have made considerable advances in the last decade, sample preparation has remained an important challenge within the workflow due to the multi-step nature of complex biological samples, and still requires much development. Several sample preparation methods have been developed and used in the last two decades, including in-gel, in-solution, on-bead, filter-aided sample preparation, and suspension trapping, to improve reproducibility, efficiency, scalability, and reduce the handling time of this process. One of the most recent methods developed and applied in proteomics studies in recent years is suspension trapping, which combines rapid detergent removal, reactor-type protein digestion, and peptide clean-up in a tip or spin column. Suspension trapping is a simple, rapid, and reproducible digestion method that can effectively handle proteins in low microgram or sub-microgram amounts. This review discusses the benefits of the suspension trapping digestion method in relation to its development and application in bottom-up proteomics studies. We also discuss recent applications of suspension trapping digestion to different sample types and the features of the suspension trapping digestion method compared with other sample preparation methods.

Cyanobacteria, as photosynthetic bacteria, can generate energy and carbon sources directly from sunlight and atmospheric carbon dioxide. With the aid of genetic engineering, heterologous enzymes and/or pathways have been introduced into cyanobacteria, enabling the production of various biofuels and biochemicals. By utilizing cyanobacteria as production hosts, biofuels and biochemicals can be produced directly from carbon dioxide, and ultimately carbon dioxide fixation by cyanobacteria can help reduce atmospheric carbon dioxide concentration, alleviating global warming and air pollution. In this review, we introduce various biofuels and biochemicals produced, particularly from acetyl-CoA by cyanobacteria, and summarize research strategies that have been made to improve their production. This review will provide comprehensive information and valuable insights into strategies for enhancing cyanobacterial biofuels/chemicals production.

Cosmic Ray Energetics And Mass for the International Space Station (ISS-CREAM) was designed to study high-energy cosmic rays up to PeV and recorded data from August 22nd, 2017 to February 12th, 2019 on the ISS. In this analysis, the Silicon Charge Detector (SCD), CALorimeter (CAL), and Top and Bottom Counting Detectors (TCD/BCD) are used. The SCD is composed of four layers and provides the measurement of cosmic-ray charges with a resolution of $\sim$0.2e. The CAL comprises 20 interleaved tungsten plates and scintillators, measures the incident cosmic-ray particles' energies, and provides a high energy trigger. The TCD/BCDs consist of photodiode arrays and plastic scintillators and provide a low-energy trigger. In this analysis, the SCD top layer is used for charge determination. Here, we present the heavy nuclei analysis using the ISS-CREAM instrument.

One of the consequences of having a high-density calorimeter as part of an experiment is a large number of secondary shower particles generated in the calorimeter -- some of which scatter back up towards the charge measurement devices. This so-called "backscatter effect" can interfere severely with accurate charge measurement of the primary nucleus, especially at high energies, as the number of backscattered particles increases with the incident energy. In this analysis, we study the effect of backscattered particles on particle identification by simulating the ISS-CREAM instrument model detector response using the GEANT3 simulation package [1] with the FLUKA hadronic model [2]. Our study shows the importance of the fine segmentation of charge detectors above the calorimeter. It can minimize backscattered particle contamination in the same charge detector segment as the incident particle to avoid its charge misidentification. We present simulation results regarding charge measurements, including the tracking resolution, backscattering effects, and charge determination efficiency.

The Cosmic Ray Energetics And Mass for the International Space Station (ISS-CREAM) experiment successfully recorded the data for about 539 days from August 2017 to February 2019. In this talk, we report the measurement of the cosmic-ray proton energy spectrum from the ISS-CREAM experiment in the energy range of 2.5 TeV - 650 TeV. For the analysis, we used the silicon charge detector (SCD) placed at the top of the ISS-CREAM payload to identify the incoming cosmic-ray charge. The SCD is finely segmented to minimize charge misidentification due to backscatter effects. The four-layer SCD consists of 10,752 silicon pixels, each of which is 1.37×1.57×0.05 cm^3 in size. The calorimeter (CAL) consists of 20 layers of tungsten/scintillating fibers preceded by carbon targets. It provided cosmic-ray tracking, energy determination, and the high-energy trigger. The Top and Bottom Counting detectors (T/BCD) are above and below the CAL, respectively, and provided the low energy trigger. Each T/BCD is composed of an array of 20×20 photodiodes on plastic scintillators. The measured proton spectral index of 2.67±0.02 between 2.5 and 12.5 TeV is consistent with prior CREAM measurements. The spectrum softens above∼10 TeV consistent with the bump-like structure as reported by CREAM-I+III, DAMPE, and NUCLEON, but ISS-CREAM extends measurements to higher energies than those prior measurement

A life cycle CO2 assessment is performed to evaluate the net CO2 reduction performance of a modeled ammonia-based carbon capture and storage (CCS) system and to identify high CO2 emitting processes to be improved with priority. The CCS system sequesters only 64% of the CO2 emitted from a power plant, which is significantly lower than the CO2 capture performance (95%), because CO2 equal to the gap is newly emitted due to energy and materials consumptions for CCS operations. The priority processes are CO2 capture, transport, and wastewater treatment. This study provides valuable information needed to develop high performance CCS systems.

We present a permanent magnet–based spherical wheel motor that can be used in omnidirectional mobility applications. The proposed motor consists of a ball‐shaped rotor with a magnetic dipole and a hemispherical shell with circumferential air‐core coils attached to the outer surface acting as a stator. Based on the rotational symmetry of the rotor poles and stator coils, we are able to model the rotor poles and stator coils as dipoles. A simple physical model constructed based on a torque model enables fast numerical simulations of motor dynamics. Based on these numerical simulations, we test various control schemes that enable constant‐speed rotation along arbitrary axes with small rotational attitude error. Torque analysis reveals that the back electromotive force induced in the coils can be used to construct a control scheme that achieves the desired results. Numerical simulations of trajectories confirm that even without explicit methods for correcting the rotational attitude error, it is possible to drive the motor with a low attitude error (

OBJECTIVES: To investigate the effect of optic disc size on correlation between Bruch’s membrane opening-minimum rim width (BMO-MRW) and peripapillary retinal nerve fibre layer (RNFL) thickness from three scan circles. METHODS: In this retrospective, observational study, non-glaucomatous eyes without visible RNFL defect or visual field loss were included. A total of 101 subjects were distributed into three groups based on disc size: group 1 (n = 26), small disc (disc area 2.43 mm(2)). All patients underwent standard ophthalmic examinations including confocal scanning laser tomography and spectral-domain optical coherence tomography. RESULTS: Global BMO-MRW was the thickest in group 1 (314.96 ± 60.38 μm, BMO area: 1.72 ± 0.45 mm(2)), followed by that in group 2 (259.03 ± 40.04 μm, BMO area: 2.29 ± 0.31 mm(2)). It was the thinnest in group 3 (236.74 ± 31.21 μm, BMO area: 2.91 ± 0.31 mm(2); p 0.05). CONCLUSIONS: Correlation between BMO-MRW values and RNFL thickness differed significantly according to disc size. Thus, when we assess BMO-MRW in relation to RNFL thickness, disc size may need to be taken into consideration.

We propose a substrate with high thermal conductivity, manufactured by the low‐temperature co‐fired ceramic (LTCC) multilayer circuit process technology, as a new DC/DC converter platform for power electronics applications. We compare the reliability and power conversion efficiency of a converter using the LTCC substrate with the one using a conventional printed circuit board (PCB) substrate, to demonstrate the superior characteristics of the LTCC substrates. The power conversion efficiencies of the LTCC‐ and PCB‐based synchronous buck converters are 95.5% and 94.5%, respectively, while those of nonsynchronous buck converters are 92.5% and 91.3%, respectively, at an output power of 100 W. To verify the reliability of the LTCC‐based converter, two types of tests were conducted. Storage temperature tests were conducted at −20 °C and 85 °C for 100 h each. The variation in efficiency after the tests was less than 0.3%. A working temperature test was conducted for 60 min, and the temperature of the converter was saturated at 58.2 °C without a decrease in efficiency. These results demonstrate the applicability of LTCC as a substrate for power conversion systems.

Cellulose is one of the constituents of plants and is an environmentally friendly resource with a semi-permanent supply and high biodegradability. CNF from cellulose is useful for various purposes. CNF has high aspect ratio and high specific surface area and it is excellent in mechanical properties and thermal stability, and it has shear-thinning flow characteristics. When CNF is coated on paper, it provides high barrier property with excellent network forming ability. The surface roughness of the coated paper is one of the characteristics that determine the quality of coated paper and printing. The surface roughness is measured by the stylus profiling, the air leak, and the optical image analysis methods. Each analysis method has certain characteristics, so surface roughness of coated paper was analyzed by those three methods and results were compared. After mixed coating of CNF and oxidized starch, the degree of roughness was increased in the probe method and the air leakage method when the coating weight and the amount of CNF were increased. The degree of roughness which is measured by Optitopo decreased when the mixed solution was coated. Because the range of roughness measured by each measurement method is different, it might give different tendencies.

The rapid and reproducible preparation of biological samples is an essential requirement for LC-MS-based clinical metabolomic profiling. Herein, we proposed a novel, automated sample preparation method for serum metabolite LC-MS analysis. This method included a one-step double-filtration process with two filters (30k and 10k MWCO). High sample throughput (up to 96 samples) was achieved in an automated manner by incorporating the process in a liquid handling robotic system. The effectiveness of the developed method was evaluated through comparison with a conventional manual centrifugation-based method using seven human serum samples. A higher metabolite recovery and better reproducibility were achieved in quantitative analysis using the automated double-filtration method. The results suggested that the multiplexed serum sample preparation system can be applied to large-scale untargeted metabolomics studies.

Cyanobacteria are promising microbial hosts for the production of diverse biofuels and biochemicals. However, compared to other model microbial hosts such as Escherichia coli and yeast, it takes a long time to genetically modify cyanobacteria. One way to efficiently engineer cyanobacteria while minimizing genetic engineering would be to develop a fast, high-throughput prototyping tool for cyanobacteria. In this study, we developed a CRISPR/Cas12a-based assay coupled with cyanobacteria cell-free systems to rapidly prototype promoter characteristics. Using this newly developed assay, we demonstrated cyanobacteria cell-free transcription for the first time and confirmed a positive correlation between the in vitro and in vivo transcription performance. Furthermore, we generated a synthetic promoter library and evaluated the characteristics of promoter subregions by using the assay. Varied promoter strength derived from random mutations were rapidly and effectively measured in a high-throughput way. We believe that this study offers an easily applicable and rapid prototyping platform to characterize promoters for cyanobacterial engineering.

In this study, magnetic steel slag biochar (MSSB) was synthesized from low-cost steel slag waste to investigate the effectiveness of steel slag biochar composite for NH4-N removal and magnetic properties in aqueous solution. The maximum adsorption capacity of NH4-N by MSSB was 4.366 mg/g according to the Langmuir model. The magnetic properties of MSSB indicated paramagnetic behavior and a saturation magnetic moment of 2.30 emu/g at 2 Tesla. The NH4-N adsorption process was well characterized by the pseudo-second order kinetic model and Temkin isotherm model. This study demonstrated the potential of magnetic biochar synthesized from steel slag waste for NH4-N removal in aqueous solution.

BackgroundTear deficiency due to lacrimal gland (LG) dysfunction is one of the major causes of dry eye disease (DED). Therefore, LG stem cell-based therapies have been extensively reported to regenerate injured lacrimal tissue; however, the number of stem cells in the LG tissue is low, and 2D long-term cultivation reduces the differentiation capacity of stem cells. Nevertheless, 3D LG organoids could be an alternative for a DED therapy because it is capable of prolonged growth while maintaining the characteristics of the LG tissue. Here, we report the development of LG organoids and their application as cell therapeutics.MethodsDigested cells from human LG tissue were mixed with Matrigel and cultured in five different media modified from human prostate/salivary organoid culture media. After organoid formation, the growth, specific marker expression, and histological characteristics were analyzed to authenticate the formation of LG organoids. The secretory function of LG organoids was confirmed through calcium influx or proteomics analysis after pilocarpine treatment. To explore the curability of the developed organoids, mouse-derived LG organoids were fabricated and transplanted into the lacrimal tissue of a mouse model of DED.ResultsThe histological features and specific marker expression of LG organoids were similar to those of normal LG tissue. In the pilocarpine-treated LG organoid, levels of internal Ca2+ions and β-hexosaminidase, a lysosomal protein in tear fluid, were increased. In addition, the secreted proteins from pilocarpine-treated lacrimal organoids were identified through proteomics. More than 70% of the identified proteins were proven to exosome through gene ontology analysis. These results indicate that our developed organoid was pilocarpine reactive, demonstrating the function of LG. Additionally, we developed LG organoids from patients with Sjogren’s syndrome patients (SS) and confirmed that their histological features were similar to those of SS-derived LG tissue. Finally, we confirmed that the mouse LG organoids were well engrafted in the lacrimal tissue two weeks after transplantation.ConclusionThis study demonstrates that the established LG organoids resemble the characteristics of normal LG tissue and may be used as a therapy for patients with DED.

AIM: To confirm the changes in proteins related with hypoxia-induced retinal cell death and to assess the effects of resveratrol (Res). METHODS: The therapeutic effect of Res was verified using an ischemic/reperfusion (I/R) model in vivo and a hypoxia modelin retinal ganglion cells (RGCs) in vitro. Death of RGCs were confirmed by TUNEL assay. Protein expression was confirmed by Western blotting and immunohistochemistry. In addition, flow cytometric analysis was used to confirm the response in the cell unit to obtain more accurate data. RESULTS: ErbB2 expression and apoptosis in the ganglion cell layer (GCL) increased after I/R injury. Treatment of Res rescued I/R-induced ganglion cell death, downregulated apoptosis and ErbB2 protein expression in the retina. In subsequent in vitro models, Res affects apoptosis by regulating the phosphorylation and expression of mouse double minute 2 homolog (MDM2), along with those of ErbB2. These results suggest that Res reverses GCL-specific apoptosis via downregulation of ErbB2 in ischemic injury. CONCLUSION: In light of Res favorable properties, it should be evaluated in the treatment of RGC death and related retinal disease characterized by ErbB2 and MDM2 expression. Therefore, Res is appropriate therapeutic agent for treating ischemic injury-related eye diseases by targeting the expression of ErbB2 and MDM2.

Additional file 1: Table S1. The components of culture medium for lacrimal gland organoid. Table S2. The primers used for quantitative RT-PCR.

Background Stem cell transplantation has been proposed as an alternative treatment for intractable optic nerve disorders characterized by irrecoverable loss of cells. Mesenchymal stem cells, with varying tissue regeneration and recovery capabilities, are being considered for potential cell therapies. To overcome the limitations of cell therapy, we isolated exosomes from human placenta-derived mesenchymal stem cells (hPMSCs) and investigated their therapeutic effects in R28 cells (retinal precursor cells) exposed to CoCl2. Method After 9 h of exposure to CoCl2, the hypoxic damaged R28 cells were divided into the non-treatment group (CoCl2 + R28 cells) and treatment group (CoCl2 + R28 cells treated with exosome). Immunoblot analysis was performed for Pcna, Hif-1α, Vegf, Vimentin, Thy-1, Gap43, Ermn, Neuroflament, Wnt3a, β-catenin, phospo-GSK3β, Lef-1, UBA2, Skp1, βTrcp, and ubiquitin. The proteomes of each group were analyzed by liquid chromatography/tandem mass (LC-MS/MS) spectrometry. Differentially expressed proteins (DEPs) were detected by label-free quantification, and the interactions of the proteins were examined through signal transduction pathway and gene ontology analysis. Result We observed that exosome could significantly recover proliferation damaged by CoCl2 treatment. In addition, the treatment group presented the decreased expression of Hif-1α protein (P P p 2 in lacking UBA2 R28 cells. Conclusion This study reported that hypoxic damaged expression of regeneration markers in R28 cells was significantly recovered by hPMSC exosomes. We could also demonstrate that UBA2 played a key role in activating the Wnt/β-catenin signaling pathway during protection of hypoxic damaged R28 cells, induced by hPMSC exosomes.