Molecular insight into the potential functional role of pseudoenzyme GFOD1 via interaction with NKIRAS2.

The glucose-fructose oxidoreductase/inositol dehydrogenase/rhizopine catabolism protein (Gfo/Idh/MocA) family includes a variety of oxidoreductases with a wide range of substrates that utilize NAD or NADP as redox cofactor. Human contains two members of this family, namely glucose-fructose oxidoreductase domain-containing protein 1 and 2 (GFOD1 and GFOD2). While GFOD1 exhibits low tissue specificity, it is notably expressed in the brain, potentially linked to psychiatric disorders and severe diseases. Nevertheless, the specific function, cofactor preference, and enzymatic activity of GFOD1 remain largely unknown. In this work, we find that GFOD1 does not bind to either NAD or NADP. Crystal structure analysis unveils that GFOD1 exists as a typical homodimer resembling other family members, but lacks essential residues required for cofactor binding, suggesting that it may function as a pseudoenzyme. Exploration of GFOD1-interacting partners in proteomic database identifies NK-κB inhibitor-interacting Ras-like 2 (NKIRAS2) as one potential candidate. Co-immunoprecipitation (co-IP) analysis indicates that GFOD1 interacts with both GTP- and GDP-bound forms of NKIRAS2. The predicted structural model of the GFOD1-NKIRAS2 complex is validated in cells using point mutants and shows that GFOD1 selectively recognizes the interswitch region of NKIRAS2. These findings reveal the distinct structural properties of GFOD1 and shed light on its potential functional role in cellular processes.

MRI analysis of and factors related to knee injuries in amateur marathon runners.

BACKGROUND: Marathons are the most challenging form of running, and amateur athletes may be more prone to injury due to a lack of professional knowledge and instruction in running. PURPOSE: To analyze the MRI manifestations of and factors related to knee injuries in amateur marathon runners. SUBJECTS: Data were collected from a hospital database of 105 qualified amateur marathon athletes (65 males,40 females), between May 2018 and December 2021. FIELD STRENGTH/SEQUENCE: 1.5T MR: sagittal fs-PDWI, sagittal T1WI and sagittal 3D-DESS sequence. ASSESSMENT: The MRI manifestations of knee joint injury were analyzed and evaluated by two radiologists. STATISTICAL TESTS: The inter-observer agreement on MRI readings was analyzed using the kappa coefficient, and binary logistic regression analysis was employed to identify factors associated with knee injuries. RESULTS: The overall prevalence of knee cartilage lesions, meniscus lesions and bone marrow edema among amateur marathon runners was 45.7%, 72.4%, and 49.5% respectively. Our analysis revealed that older age (OR = 1.135, P<0.001), higher BMI (OR = 1.236, P = 0.044), and slower pace (OR = 2.305, P = 0.017) were associated with increased risk of articular cartilage disease. Furthermore, older age (OR = 1.425, P<0.001) was identified as a risk factor for meniscal lesions, while older age (OR = 1.088, P = 0.002) was bone marrow edema. Notably, no significant correlation was observed between knee joint injuries of amateur marathon athletes and gender or the monthly running distance (P>0.05). CONCLUSIONS: The occurrence of knee injuries among amateur marathon athletes was highly prevalent, with the patellofemoral joint cartilage and posterior horn of medial meniscus being frequently affected areas. Moreover, age, BMI, running years and pace were significant risk factors of knee joint injury.

Biochemical and structural characterization of the DNA-binding properties of human TRIP4 ASCH domain reveals insights into its functional role.

TRIP4 is a conserved transcriptional coactivator that is involved in the regulation of the expression of multiple genes. It consists of a classical N-terminal C2HC5-like zinc-finger domain and a conserved C-terminal ASCH domain. Here, we characterized the DNA-binding properties of the human TRIP4 ASCH domain. Our biochemical data show that TRIP4-ASCH has comparable binding affinities toward ssDNA and dsDNA of different lengths, sequences, and structures. The crystal structures reveal that TRIP4-ASCH binds to DNA substrates in a sequence-independent manner through two adjacent positively charged surface patches: one binds to the 5'-end of DNA, and the other binds to the 3'-end of DNA. Further mutagenesis experiments and binding assays confirm the functional roles of key residues involved in DNA binding. In summary, our data demonstrate that TRIP4-ASCH binds to the 5' and 3'-ends of DNA in a sequence-independent manner, which will facilitate further studies of the biological function of TRIP4.

Effects of probiotic supplementation on 12 min run performance, mood management, body composition and gut microbiota in amateur marathon runners: A double-blind controlled trial.

Background: Probiotic supplementation has a positive effect on endurance exercise performance and body composition in athletes, but the underlying mechanisms remain unclear. Gut microbiota can provide measurable markers of immune function in athletes, and microbial composition analysis may be sensitive enough to detect stress and metabolic disorders caused by exercise. Methods: Nineteen healthy active amateur marathon runners (15 male and 4 female) with a mean age of 29.11 years volunteered to participate in this double-blind controlled study. Based on the performance of the Cooper 12-min running test (CRT), the participants were allocated into two groups to receive either a probiotic formulation comprising lactobacillus acidophilus and bifidobacterium longum (n = 10) or placebo containing maltodextrin (n = 9) for five weeks. Consistency of diet and exercise was ensured throughout the experimental period. Before and after the intervention, all participants were assessed for CRT, emotional stability and gastrointestinal symptoms, gut microbiota composition, body composition and magnetic resonance imaging (MRI) indicators of skeletal muscle microcirculation. Results: Compared to before the intervention, the probiotics group showed an increase in CRT score (2.88 ± 0.57 vs 3.01 ± 0.60 km, P＜0.05), significant improvement in GSRS and GIQLI (9.20 ± 4.64 vs 7.40 ± 3.24, 118.90 ± 12.30 vs 127.50 ± 9.85, P＜0.05), while these indicators remained unchanged in the control group, with a significant time-group interaction effect on gastrointestinal symptoms. Additionally, some MRI metabolic cycling indicators of the thigh skeletal muscle also changed in the probiotics group (P＜0.05). Regarding microbiota abundance, the probiotics group exhibited a significant increase in the abundance of beneficial bacteria and a significant decrease in the abundance of harmful bacteria post-intervention (P＜0.05). Conclusion: As a sports nutritional supplement, probiotics have the potential to improve athletic performance by optimizing the balance of gut microbiota, alleviating gastrointestinal symptoms.

Longitudinal manipulation of local nonseparability in vector beams.

The inherent nonseparability of vector beams presents a unique opportunity to explore novel optical functionalities, expanding new degrees of freedom for optical information processing. In this Letter, we introduce a novel, to the best of our knowledge, method for tailoring the local nonseparability along the propagation axis of vector beams. Employing higher-order Bessel vector beams, the longitudinal control over the local nonseparability is achieved through targeted amplitude modulation of constituent orthogonal polarization components within the main ring region. Experimental demonstrations of diverse longitudinal nonseparability profiles corroborate the efficacy and versatility of our approach, opening avenues for further exploration of the nonseparability manipulation in vector beams.

Achieving focal invariance in different background refractive indices through a dual-environment metalens.

A conventional metalens is designed with a fixed working environment, and its focal length depends on the background refractive index. In this study, we propose a dual-environment metalens that can maintain the same focal length in both media of air and water. The metalens consists of 16 types of meta-atoms with different geometries, which can cover the 0-2π phase range in both air and water. We perform finite-difference time-domain simulations to investigate the metalens and demonstrate that its focal length remains unchanged, regardless of whether the background medium is air or water. Furthermore, we investigated the optical forces within the focal field of the metalens in both air and water, indicating its potential trapping capability in these media. Our method provides a new insight into dual-environment metasurfaces and advances the methodology of electromagnetic structures in extensive applications.

Biochemical properties of sheep colostrum and its potential benefits for lamb survival: a review.

Colostrum is the initial secretion of the mammary glands following parturition, which offers main food, protection, and biological active substances for the new born. The most threatening episode of neonate's life is the initial two weeks after birth. This period is associated with high neonatal mortality and morbidity. These worthwhile losses lead to a poor prolificacy rate, low profitability, and ultimately poor performance in animal production. Hence, both diseases and mortality cause valuable losses in terms of production and economic losses. The survival of neonate is correlated with their immune status and passive immune transfer (PIT). Colostrum provides the primary source of nutrition and immunity (PIT) that protects neonates against infections. It must be given as soon as possible after birth since its immunoglobulins are absorbed within the first 16-27 hours after birth, ideally within 2-4 hours. As a result, immunoglobulin (PIT) is the most important component of distressing infectious immunity, and a passable concentration of immunoglobulin in the blood of newborn lambs is linked to their health and survival rate. In this review, we summarized the importance of colostrum in early life and its association with neonatal lamb's survival, profitability and productivity of sheep farming.

Realizing multi-function absorptions through arbitrary octagonal meta-atoms.

Metasurface absorbers (MA) typically exhibit a single type of absorption function due to their regular structures. In this study, we propose an irregular MA structure with octagonal meta-atoms. The presence of eight vertices in each meta-atom allows for tunable coordinates and offers a multitude of degrees of freedom in terms of geometry. As a result, the proposed MA exhibits diverse functionalities, including perfect absorption, multi-peaks absorption, and high absorption with a filtering window. To predict the geometric parameters of the MA structure based on a given target absorption spectrum, as well as the inverse design of the structure using the absorption spectrum as input, we employ a deep neural network combined with the particle swarm optimization algorithm. Remarkably, the mean-square error for spectrum prediction and inverse design of the MA structure is found to be as low as 0.0008 and 0.0031, respectively. This study opens up new possibilities for designing irregular electromagnetic structures and holds great potential for applications in multifunctional metasurfaces and metamaterials.

Structure and biochemical characterization of l-2-hydroxyglutarate dehydrogenase and its role in the pathogenesis of l-2-hydroxyglutaric aciduria.

l-2-hydroxyglutarate dehydrogenase (L2HGDH) is a mitochondrial membrane-associated metabolic enzyme, which catalyzes the oxidation of l-2-hydroxyglutarate (l-2-HG) to 2-oxoglutarate (2-OG). Mutations in human L2HGDH lead to abnormal accumulation of l-2-HG, which causes a neurometabolic disorder named l-2-hydroxyglutaric aciduria (l-2-HGA). Here, we report the crystal structures of Drosophila melanogaster L2HGDH (dmL2HGDH) in FAD-bound form and in complex with FAD and 2-OG and show that dmL2HGDH exhibits high activity and substrate specificity for l-2-HG. dmL2HGDH consists of an FAD-binding domain and a substrate-binding domain, and the active site is located at the interface of the two domains with 2-OG binding to the re-face of the isoalloxazine moiety of FAD. Mutagenesis and activity assay confirmed the functional roles of key residues involved in the substrate binding and catalytic reaction and showed that most of the mutations of dmL2HGDH equivalent to l-2-HGA-associated mutations of human L2HGDH led to complete loss of the activity. The structural and biochemical data together reveal the molecular basis for the substrate specificity and catalytic mechanism of L2HGDH and provide insights into the functional roles of human L2HGDH mutations in the pathogeneses of l-2-HGA.

Single-shot measurement of the Jones matrix for anisotropic media using four-channel digital polarization holography.

Dynamic measurement of the Jones matrix is crucial in investigating polarization light fields, which have wide applications in biophysics, chemistry, and mineralogy. However, acquiring the four elements of the Jones matrix instantly is difficult, hindering the characterization of random media and transient processes. In this study, we propose a single-shot measurement method of the Jones matrix for anisotropic media called "four-channel digital polarization holography" (FC-DPH). The FC-DPH system is created by a slightly off-axis superposition of reference light waves, which are modulated by a spatial light modulator (SLM), and signal light waves that pass through a Ronchi grating. The SLM enables flexible adjustment of the spatial carrier frequency, which can be adapted to different anisotropic media. The four elements of the Jones matrix can be obtained from the interferogram through the inverse Fourier transform. Optical experiments on anisotropic objects validate the feasibility and accuracy of the proposed method.

Lactate dehydrogenase D is a general dehydrogenase for D-2-hydroxyacids and is associated with D-lactic acidosis.

Mammalian lactate dehydrogenase D (LDHD) catalyzes the oxidation of D-lactate to pyruvate. LDHD mutations identified in patients with D-lactic acidosis lead to deficient LDHD activity. Here, we perform a systematic biochemical study of mouse LDHD (mLDHD) and determine the crystal structures of mLDHD in FAD-bound form and in complexes with FAD, Mn2+ and a series of substrates or products. We demonstrate that mLDHD is an Mn2+-dependent general dehydrogenase which exhibits catalytic activity for D-lactate and other D-2-hydroxyacids containing hydrophobic moieties, but no activity for their L-isomers or D-2-hydroxyacids containing hydrophilic moieties. The substrate-binding site contains a positively charged pocket to bind the common glycolate moiety and a hydrophobic pocket with some elasticity to bind the varied hydrophobic moieties of substrates. The structural and biochemical data together reveal the molecular basis for the substrate specificity and catalytic mechanism of LDHD, and the functional roles of mutations in the pathogenesis of D-lactic acidosis.

Molecular basis for the functional roles of the multimorphic T95R mutation of IRF4 causing human autosomal dominant combined immunodeficiency.

Interferon regulatory factor 4 (IRF4) is a transcription factor that regulates the development and function of immune cells. Recently, a new multimorphic mutation T95R was identified in the IRF4 DNA-binding domain (DBD) in patients with autosomal dominant combined immune deficiency. Here, we characterized the interactions of the wild-type IRF4-DBD (IRF4-DBDWT) and T95R mutant (IRF4-DBDT95R) with a canonical DNA sequence and several noncanonical DNA sequences. We found that compared to IRF4-DBDWT, IRF4-DBDT95R exhibits higher binding affinities for both canonical and noncanonical DNAs, with the highest preference for the noncanonical GATA sequence. The crystal structures of IRF4-DBDWT in complex with the GATA sequence and IRF4-DBDT95R in complexes with both canonical and noncanonical DNAs were determined, showing that the T95R mutation enhances the interactions of IRF4-DBDT95R with the canonical and noncanonical DNAs to achieve higher affinity and specificity. Collectively, our data provide the molecular basis for the gain-of-function and new function of IRF4T95R.

Spatio-temporal structuring control of a vectorial focal field.

Focal field modulation has attracted a lot of interest due to its potential in many applications such as optical tweezers or laser processing, and it has recently been facilitated by spatial light modulators (SLMs) owing to their dynamic modulation abilities. However, capabilities for manipulating focal fields are limited by the space-bandwidth product of SLMs. This difficulty can be alleviated by taking advantage of the high-speed modulation ability of digital micromirror devices (DMDs), i.e., trading time for space to achieve fine focus shaping. In this paper, we propose a new, to the best of our knowledge, technique for achieving four-dimensional focal field modulation, which allows for independent manipulation of the focal field's parameters (including amplitude, phase, and polarization) in both the space and time domains. This technique combines a DMD and a vector field synthesis system based on a 4-f system. The high-speed modulation ability of DMDs enables versatile focus patterns to be fast switchable during the exposure time of the detector, forming multiple patterns in a single recording frame. By generating different kinds of focal spots and lines at different moments during the exposure time of the detector, we can finally get complete multifocal spots and lines. Our proposed method is effective at improving the flexibility and speed of the focal field modulation, which is beneficial to applications.

Molecular insights into the catalysis and regulation of mammalian NAD-dependent isocitrate dehydrogenases.

Eukaryotic NAD-dependent isocitrate dehydrogenases (NAD-IDHs) are mitochondria-localized enzymes which catalyze the oxidative decarboxylation of isocitrate to α-ketoglutarate using NAD as a cofactor. In mammals, NAD-IDHs (or IDH3) consist of three types of subunits (α, ß, and γ), and exist as (α2ßγ)2 heterooctamer. Mammalian NAD-IDHs are regulated allosterically and/or competitively by a diversity of metabolites including citrate, ADP, ATP, NADH, and NADPH, which are associated with cellular metabolite flux, energy demands, and redox status. Proper assembly of the component subunits is essential for the catalysis and regulation of the enzymes. Recently, crystal structures of human IDH3 have been solved in apo form and in complex with various ligands, revealing the molecular mechanisms for the assembly, catalysis, and regulation of the enzyme.

Protein-metabolite interactomics of carbohydrate metabolism reveal regulation of lactate dehydrogenase.

Metabolic networks are interconnected and influence diverse cellular processes. The protein-metabolite interactions that mediate these networks are frequently low affinity and challenging to systematically discover. We developed mass spectrometry integrated with equilibrium dialysis for the discovery of allostery systematically (MIDAS) to identify such interactions. Analysis of 33 enzymes from human carbohydrate metabolism identified 830 protein-metabolite interactions, including known regulators, substrates, and products as well as previously unreported interactions. We functionally validated a subset of interactions, including the isoform-specific inhibition of lactate dehydrogenase by long-chain acyl-coenzyme A. Cell treatment with fatty acids caused a loss of pyruvate-lactate interconversion dependent on lactate dehydrogenase isoform expression. These protein-metabolite interactions may contribute to the dynamic, tissue-specific metabolic flexibility that enables growth and survival in an ever-changing nutrient environment.

The Accuracy and Clinical Relevance of the Multi-echo Dixon Technique for Evaluating Changes to Hepatic Steatosis in Patients with Non-alcoholic Fatty Liver Disease Treated with Formulated Food.

PURPOSE: The Multi-echo Dixon (ME-Dixon) is a non-invasive quantitative MRI technique to diagnose non-alcoholic fatty liver disease (NAFLD). In this study, the hydrogen proton MR spectroscopy (1H-MRS) was used as a reference to explore the accuracy of the ME-Dixon technique in evaluating hepatic steatosis in NAFLD patients after ingesting formulated food and its correlation with changes in clinical indicators. METHODS: Twenty-seven patients with NAFLD were enrolled. Fifteen patients completed 12 weeks of treatment with prebiotics and dietary fiber. In addition, abdominal MRI scans and blood tests were performed before and after treatment. The MRI-proton density fat fraction (MRI-PDFF) and MRS-PDFF were measured using the ME-Dixon and 1H-MRS techniques. The Bland-Altman method and Pearson correlation analysis were used to test the consistency of the two techniques for measuring the liver fat content and the changed values. Besides, correlation analysis was conducted between the MRI-PDFF value and metabolic indicators. RESULTS: In the PDFF quantification of 42 person-times and the monitoring of the PDFF change in 15 patients under treatment, there was a good consistency and a correlation between MRI and MRS. At baseline, MRI-PDFF was positively correlated with insulin resistance index (HOMA-IR), fatty liver index (FLI), and liver enzymes. After treatment, the changes in MRI-PDFF were positively correlated with the recovery degree of FLI and liver enzymes. CONCLUSION: ME-Dixon has a good consistency and a correlation with MRS in quantifying the liver fat content and monitoring the treatment effect, which may be used as an accurate indicator for clinical monitoring of changes in the liver fat content.

Breaking symmetry restriction of chirality through spin-decoupled phase modulation utilizing non-mirror-symmetric meta-atoms.

The geometric phase in metasurfaces follows a symmetry restriction of chirality, which dictates that the phases of two orthogonal circularly polarized waves are identical but have opposite signs. In this study, we propose a general mechanism to disrupt this symmetric restriction on the chirality of orthogonal circular polarizations by introducing mirror-symmetry-breaking meta-atoms. This mechanism introduces a new degree of freedom in spin-decoupled phase modulation without necessitating the rotation of the meta-atom. To demonstrate the feasibility of this concept, we design what we believe is a novel meta-atom with a QR-code structure and successfully showcase circular-polarization multiplexing metasurface holography. Our investigation offers what we believe to be a novel understanding of the chirality in geometric phase within the realm of nanophotonics. Moreover, it paves the way for the development of what we believe will be novel design methodologies for electromagnetic structures, enabling applications in arbitrary wavefront engineering.

Structures of a constitutively active mutant of human IDH3 reveal new insights into the mechanisms of allosteric activation and the catalytic reaction.

Human NAD-dependent isocitrate dehydrogenase or IDH3 (HsIDH3) catalyzes the decarboxylation of isocitrate into α-ketoglutarate in the tricarboxylic acid cycle. It consists of three types of subunits (α, ß, and γ) and exists and functions as the (αßαγ)2 heterooctamer. HsIDH3 is regulated allosterically and/or competitively by numerous metabolites including CIT, ADP, ATP, and NADH. Our previous studies have revealed the molecular basis for the activity and regulation of the αß and αγ heterodimers. However, the molecular mechanism for the allosteric activation of the HsIDH3 holoenzyme remains elusive. In this work, we report the crystal structures of the αß and αγ heterodimers and the (αßαγ)2 heterooctamer containing an α-Q139A mutation in the clasp domain, which renders all the heterodimers and the heterooctamer constitutively active in the absence of activators. Our structural analysis shows that the α-Q139A mutation alters the hydrogen-bonding network at the heterodimer-heterodimer interface in a manner similar to that in the activator-bound αγ heterodimer. This alteration not only stabilizes the active sites of both αQ139Aß and αQ139Aγ heterodimers in active conformations but also induces conformational changes of the pseudo-allosteric site of the αQ139Aß heterodimer enabling it to bind activators. In addition, the αQ139AICT+Ca+NADßNAD structure presents the first pseudo-Michaelis complex of HsIDH3, which allows us to identify the key residues involved in the binding of cofactor, substrate, and metal ion. Our structural and biochemical data together reveal new insights into the molecular mechanisms for allosteric regulation and the catalytic reaction of HsIDH3.