Effects of biomass co-pyrolysis and herbaceous plant colonization on the transformation of tailings into soil like substrate.

Enhancing soil organic matter characteristics, ameliorating physical structure, mitigating heavy metal toxicity, and hastening mineral weathering processes are crucial approaches to accomplish the transition of tailings substrate to a soil-like substrate. The incorporation of biomass co-pyrolysis and plant colonization has been established to be a significant factor in soil substrate formation and soil pollutant remediation. Despite this, there is presently an absence of research efforts aimed at synergistically utilizing these two technologies to expedite the process of mining tailings soil substrate formation. The current study aimed to investigate the underlying mechanism of geochemical changes and rapid mineral weathering during the process of transforming tailings substrate into a soil-like substrate, under the combined effects of biomass co-smoldering pyrolysis and plant colonization. The findings of this study suggest that the incorporation of smoldering pyrolysis and plant colonization induces a high-temperature effect and biological effects, which enhance the physical and chemical properties of tailings, while simultaneously accelerating the rate of mineral weathering. Notable improvements include the amelioration of extreme pH levels, nutrient enrichment, the formation of aggregates, and an increase in enzyme activity, all of which collectively demonstrate the successful attainment of tailings substrate reconstruction. Evidence of the accelerated weathering was verified by phase and surface morphology analysis using X-ray diffraction and scanning electron microscopy. Discovered corrosion and fragmentation on the surface of minerals. The weathering resulted in corrosion and fragmentation of the surface of the treated mineral. This study confirms that co-smoldering pyrolysis of biomass, combined with plant colonization, can effectively promote the transformation of tailings into soil-like substrates. This method has can effectively address the key challenges that have previously hindered sustainable development of the mining industry and provides a novel approach for ecological restoration of tailings deposits.

Identification of APPB1 as a substrate for anaplastic lymphoma kinase.

Anaplastic lymphoma kinase (ALK) is a well-known oncogene involved in various malignancies such as anaplastic large cell lymphoma, lung cancer and neuroblastoma. Several substrates for fused ALK have been identified and their biological functions have been described. However, the lack of a comprehensive identification of ALK substrates limits our understanding of the biological roles of receptor ALK. Thus, this study aimed to identify novel ALK substrates and characterize their biological functions. We screened the interactors of the kinase domain of receptor ALK using proximity-dependent biotin identification and identified 43 interactors. We narrowed down the candidates by evaluating whether these interactors were downstream of ALK in a neuroblastoma cell line, NB-1. Among these, we identified amyloid beta precursor protein binding family B member 1 (APBB1) as an ALK downstream molecule involved in NB-1 cell viability. Finally, we assessed the kinase-substrate relationship between ALK and APBB1 and found that ALK phosphorylated multiple tyrosine residues in APBB1 both in-cell and in-tube assays, with tyrosine 269 as a major target. In conclusion, we successfully identified a new substrate for receptor ALK. Our results may help further elucidate the molecular mechanism of ALK downstream signaling in neuroblastoma.

Chiral Liquid-Crystal-Based Concave Holographic Spectrometer on a Curved Substrate.

Chiral liquid crystals (CLCs) self-assemble into a helical structure and can efficiently reflect circularly polarized light with corresponding handedness. Utilizing a curved glass substrate and polymerization of photoaligned CLCs, the operation of focusing and diffraction of incident light can be performed efficiently by a single component. When focusing and diffraction in a planar CLC cell are combined between two glass plates, the imaging suffers from astigmatism in the resulting spectrum. In this work, we demonstrate the operation of a spectrometer with low astigmatism using a polymerized CLC layer on a curved substrate. Two samples are fabricated, and the resulting components are operating in the wavelength range of 500-650 nm. Numerical optical modeling is used to minimize transverse aberrations and obtain a highly linear mapping on a camera sensor. In this way, it is demonstrated that a single reflective thin-film optical CLC component with a thickness of only a few micrometers can be used to realize a compact and efficient spectrometer.

[What is confirmed in the treatment of Fabry's disease?] / Was ist gesichert in der Therapie von Morbus Fabry?

Fabry's disease is a rare X chromosome-linked inherited lysosomal storage disease characterized by insufficient metabolism of the substrate globotriaosylceramide (Gb3) due to reduced alpha-galactosidase A (AGAL) activity. Lysosomal Gb3 accumulation causes a multisystemic disease which, if untreated, reduces the life expectancy in females and males by around 10 and 20 years, respectively, due to progressive renal dysfunction, hypertrophic cardiomyopathy, cardiac arrhythmia and early occurrence of cerebral infarction. The diagnosis is confirmed by determining the reduced AGAL activity in leukocytes in males and molecular genetic detection of a -mutation causing the disease in females. The treatment comprises enzyme replacement therapy (ERT), agalsidase alfa, 0.2 mg/kg body weight (BW), agalsidase beta 1.0 mg/kg BW or pegunigalsidase alfa 1.0 mg/kg BW every 2 weeks i.v. or oral chaperone therapy (one capsule of migalastat 123 mg every other day) in the presence of amenable mutations. This article summarizes the data on the treatment of Fabry's disease and on complications in practice. The current guideline recommendations are addressed and new study results that could expand the therapeutic repertoire in the future are discussed.

Structure-Assisted Design of Chitosanase Product Specificity for the Production of High-Degree Polymerization Chitooligosaccharides.

Chitosanases are valuable enzymatic tools in the food industry for converting chitosan into functional chitooligosaccharides (COSs). However, most of the chitosanases extensively characterized produced a low degree of polymerization (DP) COSs (DP = 1-3, LdpCOSs), indicating an imperative for enhancements in the product specificity for the high DP COS (DP >3, HdpCOSs) production. In this study, a chitosanase from Methanosarcina sp. 1.H.T.1A.1 (OUC-CsnA4) was cloned and expressed. Analysis of the enzyme-substrate interactions and the subsite architecture of the OUC-CsnA4 indicated that a Ser49 mutation could modify its interaction pattern with the substrate, potentially enhancing product specificity for producing HdpCOSs. Site-directed mutagenesis provided evidence that the S49I and S49P mutations in OUC-CsnA4 enabled the production of up to 24 and 26% of (GlcN)5 from chitosan, respectively─the wild-type enzyme was unable to produce detectable levels of (GlcN)5. These mutations also altered substrate binding preferences, favoring the binding of longer-chain COSs (DP >5) and enhancing (GlcN)5 production. Furthermore, molecular dynamics simulations and molecular docking studies underscored the significance of +2 subsite interactions in determining the (GlcN)4 and (GlcN)5 product specificity. These findings revealed that the positioning and interactions of the reducing end of the substrate within the catalytic cleft are crucial factors influencing the product specificity of chitosanase.

Carbon and nitrogen addition-derived enzyme activities in topsoil but nitrogen availability in subsoil controls the response of soil organic carbon decomposition to warming.

Subsoil stores the majority of soil organic carbon (SOC), and plays a vital role in the global carbon cycle in terrestrial ecosystems and in regulating climate change. Response of SOC decomposition to temperature warming (TR) is a crucial parameter to predict SOC dynamics under global warming. However, it remains unknown how TR varies across the whole soil profile and responds to exogenous C and N inputs. To assess this, we designed a novel incubation system to measure SOC-derived CO2 efflux across the whole soil column (i.e., 60 cm length), allowing manual addition of 13C-labeled glucose and ammonium nitrate, and incubated it under ambient or warmed temperatures (+4 °C). We found that C addition significantly increased TR in 0-20 cm, 20-40 cm and 40-60 cm by 64.3 %, 68.1 % and 57.2 %, respectively. However, the combined addition of C and N decreased TR by 11.1 % - 15.3 % compared to without anything addition (CK) in the whole soil profile. The effect of N on TR ranged from -22.8 % to -40.4 % in the whole soil profile, and was significantly lower in topsoil than in subsoil. Furthermore, sole N addition significantly promoted TR compared to CK by 79.0 % and 94.7 % in 20-40 cm and 40-60 cm subsoil, only 9.8 % in 0-20 cm topsoil. These results together suggested that TR is sensitive to increasing C availability in the whole soil profile and increasing N availability in 20-60 cm subsoil. Random forest model indicated that soil enzyme activities (explained 21.3 % of the variance) and DOC (explained 11.1 % of the variance) dominantly governed TR in topsoil, but N availability displayed a predominant control of TR in subsoil. Overall, our results suggested that increased C and N availability under climate warming scenarios could further increase the risk of carbon loss especially in subsoil with substrate deficiency, but labile C (e.g., root exudation) input under climate warming and N enrichment could reduce SOC decomposition and benefit for C sequestration by decreasing TR.

Machine learning-assisted substrate binding pocket engineering based on structural information.

Engineering enzyme-substrate binding pockets is the most efficient approach for modifying catalytic activity, but is limited if the substrate binding sites are indistinct. Here, we developed a 3D convolutional neural network for predicting protein-ligand binding sites. The network was integrated by DenseNet, UNet, and self-attention for extracting features and recovering sample size. We attempted to enlarge the dataset by data augmentation, and the model achieved success rates of 48.4%, 35.5%, and 43.6% at a precision of ≥50% and 52%, 47.6%, and 58.1%. The distance of predicted and real center is ≤4 Å, which is based on SC6K, COACH420, and BU48 validation datasets. The substrate binding sites of Klebsiella variicola acid phosphatase (KvAP) and Bacillus anthracis proline 4-hydroxylase (BaP4H) were predicted using DUnet, showing high competitive performance of 53.8% and 56% of the predicted binding sites that critically affected the catalysis of KvAP and BaP4H. Virtual saturation mutagenesis was applied based on the predicted binding sites of KvAP, and the top-ranked 10 single mutations contributed to stronger enzyme-substrate binding varied while the predicted sites were different. The advantage of DUnet for predicting key residues responsible for enzyme activity further promoted the success rate of virtual mutagenesis. This study highlighted the significance of correctly predicting key binding sites for enzyme engineering.

Case report of persistent atrial fibrillation with durably isolated pulmonary veins: what's next?

Background: Pulmonary vein isolation (PVI) has emerged as a safe and effective treatment for patients with paroxysmal and persistent atrial fibrillation. Nevertheless, in some patients, a relapse of atrial fibrillation occurs although pulmonary veins are durably isolated. For those patients, the underlying mechanisms of atrial fibrillation perpetuation are manifold and optimal treatment options are not yet defined. Case summary: We describe a case of a 55-year-old man with a history of atrial fibrillation and previous PVI presenting with persistent AF and arrhythmia induced cardiomyopathy. During the redo procedure, electro-anatomical mapping revealed durably isolated PV. Bipolar mapping showed large low-voltage areas at the posterior wall and the septum. As the patient was refractory to electrical cardioversion, it was decided to modify the large low-voltage areas as potential arrhythmic substrate. After performing additional ablation with isolation of the posterior wall and two anterior/septal lines, the patient spontaneously converted to sinus rhythm. Discussion: Ablation in patients with persistent AF despite durable PVI remains a challenge for the treating team. Individualized ablation approaches addressing additional arrhythmic substrates or extra-PV triggers can be considered to treat patients with persistent AF and durable PVI.

Textile-Based Membraneless Microfluidic Double-Inlet Hybrid Microbial-Enzymatic Biofuel Cell.

This study reports the development of a textile-based colaminar flow hybrid microbial-enzymatic biofuel cell. Shewanella MR-1 was used as a biocatalyst on the anode, and bienzymatic system catalysts based on glucose oxidase and horseradish peroxidase were applied on an air-breathing cathode to address the overpotential loss in a body-friendly way. A single-layer Y-shaped channel configuration with a double-inlet was adopted. Microchannels of biofuel cells were patterned by silk screen printing with Ecoflex to maintain the flexibility of textile substrates without harm to the human body. The electrodes were fabricated with poly(3,4-ethylenedioxythiophene):polystyrene sulfonate and a mixture of multiwalled carbon nanotubes and single-walled carbon nanotubes by screen printing. The effects of electrode materials, catalyst type, catalyst concentration, and glucose concentration in the catholyte were investigated to optimize the fuel cell performance. The peak power density (44.9 µW cm-2) and maximum current density (388.9 µA cm-2) of the optimized hybrid biofuel cell were better than those of previously reported textile- or paper-substrate microscale single microbial fuel cells. The developed biofuel cell will be a useful platform as a microscale power source that is harmless to the environment and living organisms.

Investigation the behavior of different fullerenes on graphene surface.

In the present study, the regime of motion of fullerene molecules on graphene substrate in a specific temperature range is investigated. The potential energy of fullerene molecules is analyzed using classical molecular dynamics methods. Fullerene molecules C36, C50, C60, C76, C80, and C90 are selected due to spherical shapes of different sizes and good motion performance in previous studies. Analysis of the motion regime at different temperatures is one of the main objectives of this study. To achieve this aim, the translational and rotational movements of fullerene molecules are studied independently. In the first step of the investigation, Lennard-Jone's potential energy of fullerene molecules is calculated. Subsequently, the motion regime of different fullerenes is classified based on their displacement and diffusion coefficient. Findings indicate C60 is not appropriate in all conditions. However, C90 and C76 molecules are found to be appropriate candidates in most cases in different conditions. As far as a straight-line movement is considered, the deviation of fullerene molecules is compared by their angular velocities. Although C60 has a lower angular velocity due to its symmetrical shape, it may not move well due to its low diffusion coefficient. Overall, our study helps to understand the performance of different fullerene molecules on graphene substrate and find their possible applications, especially as wheels in nanomachine or nanocarrier structures.

Using a metasurface to enhance the radiation efficiency of subterahertz antennas printed on thick substrates.

This study investigates the possibility of increasing the radiation efficiency of printed antennas and arrays by suppressing their inherent surface waves using a metasurface made of quad-split rings (QSR). A symmetrical resonant microstrip dipole and a four-element series-fed dipole array printed on an infinite grounded dielectric layer (layer thickness: 0.2 mm; relative permittivity: 9.4; tanδ: 0.0005) were simulated with FEKO 2022 software. Conducted at 100-116 GHz, the numerical results revealed extremely low radiation efficiencies of approximately 31% and 40% for the studied dipole and dipole array, respectively, which resulted from the presence of surface waves in the dielectric. However, placing only one QSR near each dipole arm triggered an increase in radiation efficiency by 2.5 times (up to 75%). The use of a metasurface in the form of two small QSR arrays triggered a pronounced improvement in radiation efficiency, reaching 93.6% and 95.8% for the studied dipole and dipole array, respectively. Analysis of the electric field distribution images showed that this enhancement resulted from surface wave suppression.

The roles of FHL2 as a mechanotransducer for cellular functions in the mechanical environment.

The cell has multiple mechanisms for sensing and responding to dynamic changes in the mechanical environment. In the process, intracellular signaling is activated to modulate gene expression. Recent studies have shown that multifunctional signaling molecules that link intracellular force and gene expression are important for understanding cellular functions in the mechanical environment. This review discusses recent studies on one of the mechanotransducers, Four-and-a-half LIM domains 2 (FHL2), which localizes to focal adhesions (FAs), actin cytoskeleton, and nucleus. FHL2 localizes to FAs and the actin cytoskeleton in the cell on stiff substrate. In this situation, intracellular tension of F-actin by Myosin II is critical for FHL2 localization to FAs and actin stress fibers. In the case, a conserved phenylalanine in each LIM domain is responsible for its localization to F-actin. On the other hand, lower tension of F-actin in the cell on a soft substrate causes FHL2 to be released into the cytoplasm, resulting in its localization in the nucleus. At the molecular level, phosphorylation of specific tyrosine in FHL2 by FAK, non-receptor tyrosine kinase, is critical to nuclear localization. Finally, by binding to transcription factors, FHL2 modulates gene expression for cell proliferation as a transcriptional co-factor. Thus, FHL2 is involved in mechano-sensing and -transduction in the cell in a mechanical environment.

Conductive single-phase SrMoO3 epitaxial films synthesized in pure Ar ambience via plasma-assisted radio frequency sputtering.

The cubic perovskite SrMoO3 with a paramagnetic ground state and remarkably low room-temperature resistivity has been considered as a suitable candidate for the new-era oxide-based technology. However, the difficulty of preparing single-phase SrMoO3 thin films by hydrogen-free sputtering has hindered their practical use, especially due to the formation of thermodynamically favorable SrMoO4 impurity. In this work, we developed a radio frequency sputtering technology enabling the reduction reaction and achieved conductive epitaxial SrMoO3 films with pure phase from a SrMoO4 target in a hydrogen-free, pure argon environment. We demonstrated the significance of controlling the target-to-substrate distance (TSD) on the synthesis of SrMoO3; the film resistivity drastically changes from 1.46 × 105 µΩ·cm to 250 µΩ·cm by adjusting the TSD. Cross-sectional microstructural analyses demonstrated that films with the lowest resistivity, deposited for TSD = 2.5 cm, possess a single-phase SrMoO3 with an epitaxial perovskite structure. The formation mechanism of the conductive single-phase SrMoO3 films can be attributed to the plasma-assisted growth process by tuning the TSD. Temperature-dependent resistivity and Hall effect studies revealed metal-like conducting properties for low-resistive SrMoO3 films, while the high-resistive ones displayed semiconductor-like behavior. Our approach makes hydrogen-free, reliable and cost-efficient scalable deposition of SrMoO3 films possible, which may open up promising prospects for a wide range of future applications of oxide materials.

For the first time, we developed a plasma-assisted RF sputtering technology enabling the reduction reaction for the synthesis of single-phase conductive SrMoO₃ epitaxial films from insulating SrMoO₄ in pure-argon atmosphere.

Degradation-driven vegetation-soil-microbe interactions alter microbial carbon use efficiency in Moso bamboo forests.

Microbial carbon utilization efficiency (CUE) is a crucial indicator for evaluating the efficiency of soil carbon sequestration and transformation, which is applied to quantify the proportion of soil carbon extracted by microbes for anabolism (growth) and catabolism (respiration). It has been found that the degradation of Moso bamboo forest (Phyllostachys edulis) leads to the destruction of aboveground bamboo structure, reduced vegetation carbon storage, and weakened ecosystem carbon sequestration capacity. Interestingly, soil organic carbon storage is gradually increasing. However, the mechanism by which degradation-induced changes in soil and vegetation characteristics affect microbial CUE and drive soil carbon sequestration remains unclear. Here we selected four stands with the same origin but different degradation years (intensive management, CK; 2 years' degradation, DM1; 6 years' degradation, DM2; and 10 years' degradation, DM3) based on the local management profiles. The principle of space-for-time substitution was used to investigate the changes in microbial CUE along a degradation time and to further identify the controlling biotic and abiotic factors. Our finding showed that microbial CUE increased by 12.27 %, 31.01 %, and 55.95 %, respectively, compared with CK; whereas microbial biomass turnover time decreased from 23.99 ±â€¯1.11 to 17.16 ±â€¯1.20 days. Promoting microbial growth was the main pathway to enhance microbial CUE. Massive inputs of vegetative carbon replenished soil carbon substrate content, and altered microbial communities and life history strategy, which in turn promoted microbial growth and increased microbial CUE. These findings provide theoretical support for the interactions between carbon dynamics and microbial physiology in degraded bamboo forests, and reinforce the importance of vegetation and microbial properties and soil carbon substrates in predicting microbial CUE.

Stretchable Triboelectric Nanogenerator Based on Liquid Metal with Varying Phases.

Stretchable triboelectric nanogenerators (TENGs) represent a new class of energy-harvesting devices for powering wearable devices. However, most of them are associated with poor stretchability, low stability, and limited substrate material choices. This work presents the design and demonstration of highly stretchable and stable TENGs based on liquid metalel ectrodes with different phases. The conductive and fluidic properties of eutectic gallium-indium (EGaIn) in the serpentine microfluidic channel ensure the robust performance of the EGaIn-based TENG upon stretching over several hundred percent. The bi-phasic EGaIn (bGaIn) from oxidation lowers surface tension and increases adhesion for printing on diverse substrates with high output performance parameters. The optimization of the electrode shapes in the bGaIn-based TENGs can reduce the device footprint and weight, while enhancing stretchability. The applications of the EGaIn- and bGaIn-based TENG include smart elastic bands for human movement monitoring and smart carpets with integrated data transmission/processing modules for headcount monitoring/control. Combining the concept of origami in the paper-based bGaIn TENG can reduce the device footprint to improve output performance per unit area. The integration of bGaIn-TENG on a self-healing polymer substrate with corrosion resistance against acidic and alkaline solutions further facilitates its use in various challenging and extreme environments.

Porous red mud ceramsite for aquatic phosphorus removal: Application in constructed wetlands.

Red mud (RM) and spent oyster mushroom substrate (SOMS), by-products of industrial and agricultural production, can be recycled for polluted freshwater purification, bringing about a win-win situation. In this study, unacidified RM and RM acidified with oxalic acid (O-RM) and hydrochloric acid (H-RM), respectively, were mixed with SOMS to produce a porous ceramsite as a potential constructed wetlands (CWs) substrate. The results showed that the O-RM, H-RM, and RM ceramsites displayed fine compressive strengths of 7.75 ± 1.14, 8.40 ± 1.30, and 8.84 ± 0.69 MPa after calcining at 950 °C for 30 min, respectively. The phosphorus adsorption capacities of H-RM, O-RM, and RM ceramsite at a solid-liquid ratio of 25 g/L were 1.18 mg/g, 0.88 mg/g, and 1.06 mg/g, respectively. Toxicity release experiments showed that the ceramsites did not cause secondary environmental pollution, except for arsenic (ranging from 0.210 to 0.238 mg/L). The H-RM ceramsite was tested in a tidal flow-vertical flow CW (TF-VFCW) with Iris pseudacorus L. and Canna indica L plants. In the TF-VFCW, the average chemical oxygen demand (COD), ammonia nitrogen (NH4+-N), total nitrogen (TN), and total phosphorus (TP) removal rates were 81.01, 90.25, 66.90, and 77.32 %, respectively. Plant growth had less impact on COD and NH4-N removal but had greater limited TN and TP removal. Scanning electron microscopy, Fourier-transform infrared spectroscopy, and X-ray diffraction analysis confirmed that acid pretreatment and the incorporation of SOMS significantly increased the surface and interior porous structures of the ceramsite and enhanced phosphate adsorption by the polyhydroxyl aluminum-iron complex ions. Bacteroides and Campylobacter used the energy produced during polyhydroxyalkanoic acid (PHA) catabolism to absorb phosphorus. Therefore, the synergistic effect of the substrate, plants, and microorganisms achieved the removal of phosphorus from CWs and offered effective and environmentally friendly recycling of RM and SOMS.

Substrate specificity modification of paraben hydrolase and tannase from Aspergillus oryzae.

Paraben hydrolase and tannase catalyze the hydrolysis of parabens (4-hydroxybenzoic acid esters) and gallic acid (3,4,5-trihydroxybenzoic acid) esters, respectively. Paraben hydrolase (AoPrbA) and tannase (AoTanB) from Aspergillus oryzae belong to the tannase family in the ESTHER database. However, the substrate specificities of AoPrbA and AoTanB are narrow. Based on structural information of Aspergillus niger tannase (PDB code 7k4o), we constructed five single variants of AoPrbA (Thr200Glu, Phe231Gln, Leu232Gln, Ile361Tyr, and Leu428Ser) and four of AoTanB (Glu203Asp, Glu203Thr, His237Ala, and Ser440Leu) to investigate substrate discrimination between AoPrbA and AoTanB. Each variant was expressed in Pichia pastoris and were purified from the culture supernatant. Five purified variants of AoPrbA and four variants of AoTanB showed reduced paraben hydrolase and tannase activities compared with AoPrbA and AoTanB wild types, respectively. Interestingly, the AoPrbA wild type did not hydrolyze gallic acid methyl ester, whereas the Thr200Glu, Leu232Gln, and Leu428Ser variants did, indicating that these three variants acquired tannase activity. In particular, the Leu428Ser variant exhibited considerably greater hydrolysis of gallic acid and protocatechuic acid methyl esters. Meanwhile, the AoTanB wild type, and Glu203Asp, His237Ala and Ser440Leu variants hydrolyzed the protocatechuate methyl and 4-hydroxybenzoate ethyl esters; however, the Glu203Thr variant did not hydrolyze above-mentioned substrates. Additionally, the ratio of paraben hydrolase activity to tannase activity in Ser440Leu was markedly elevated.

A structural peculiarity of Antarctic fish IgM drives the generation of an engineered mAb by CRISPR/Cas9.

IgM is the major circulating Ig isotype in teleost fish, showing in Antarctic fish unique features such as an extraordinary long hinge region, which plays a crucial role in antibody structure and function. In this work, we describe the replacement of the hinge region of a murine monoclonal antibody (mAb) with the peculiar hinge from Antarctic fish IgM. We use the CRISPR/Cas9 system as a powerful tool for generating the engineered mAb. Then, we assessed its functionality by using an innovative plasmonic substrate based on bimetallic nanoislands (AgAuNIs). The affinity constant of the modified mAb was 2.5-fold higher than that obtained from wild-type mAb against the specific antigen. Here, we show the suitability of the CRISPR/Cas9 method for modifying a precise region in immunoglobulin gene loci. The overall results could open a frontier in further structural modifications of mAbs for biomedical and diagnostic purposes.

Betaine for the prevention and treatment of insulin resistance and fatty liver in a high-fat dietary model of insulin resistance in C57BL mice.

Aim: The aim was to investigate mechanisms by which betaine improves hepatic insulin signaling in a dietary mouse model of insulin resistance and fatty liver. Methods: C57BL 6J mice were fed a standard diet (SF), a standard diet with betaine (SFB), a nutritionally complete high fat (HF) diet, or a high fat diet with betaine (HFB) for 14 weeks. In a separate experiment, mice were fed high fat diet for 18 weeks, half of whom received betaine for the final 4 weeks. Activation of insulin signaling in the liver was assessed by western blot. Insulin signaling was also assessed in insulin resistant primary human hepatocytes treated with betaine. Results: As compared with SF, mice receiving HF diet were heavier, had more hepatic steatosis, and abnormal glucose tolerance test (GTT). Betaine content in liver and serum was 50% lower in HF than in SF; betaine supplementation restored serum and liver betaine content. Betaine treatment of HF reduced whole body insulin resistance as measured by GTT. Betaine treatment of HF increased tyrosine phosphorylation of insulin receptor substrate-1 and phosphorylation (activation) of Akt, and increased hepatic glycogen content. In vitro, betaine reversed insulin resistance in primary human hepatocytes by increasing insulin-stimulated tyrosine phosphorylation of IRS1 and of Akt. Conclusion: Betaine supplementation reduced whole body insulin resistance and increased activation of insulin signaling pathways in the liver in a mouse model of insulin resistance and fatty liver created by feeding a nutritionally complete high fat diet for 14 weeks. Betaine also reduced liver injury as assessed by ALT and by liver histology. In vitro, betaine reversed insulin resistance by increasing insulin-stimulated tyrosine phosphorylation of IRS1 and activation of downstream proteins in the insulin signaling cascade in insulin resistant primary human hepatocytes.