Substrate Pairs Research Articles

Difference in structural changes of surfactant aggregates near solid surface under shear flow versus those in the bulk.

In water, the nonionic surfactant pentaethylene glycol monododecyl ether (C12E5) forms multi-lamellar vesicles upon application of shear, attributed to buckling instability of the surfactant layers. In the standard setup for applying shear, a pair of solid substrates is moved in opposite directions, and a non-slip condition at the solid surface is assumed. Based on theoretical predictions, the effective viscosity of the fluid surrounding the membrane is modified in this process, and this confinement may affect membrane fluctuation. However, only a few studies have analyzed the structural changes near the substrate. From this viewpoint, the structural changes in surfactant aggregates near a solid substrate under the application of shear were investigated herein using neutron reflectometry (NR). By increasing the shear rate, shear thickening at a lower shear rate and shear thinning at a higher shear rate were observed, similar to that in the bulk. However, a discontinuous change in the lamellar structure accompanying the condensation of the surfactant was observed in the NR experiments. This study presents the first experimental evidence indicating that the ramping speed of shear rates governs the shear-induced structuring of surfactant aggregates near the surface.

Disruptions to protein kinase A localization in adrenal pathology.

Cell signaling fidelity requires specificity in protein-protein interactions and precise subcellular localization of signaling molecules. In the case of protein phosphorylation, many kinases and phosphatases exhibit promiscuous substrate pairing and therefore require targeting interactions to modify the appropriate substrates and avoid cross-talk among different pathways. In the past 10 years, researchers have discovered and investigated how loss of specific interactions and subcellular targeting for the protein kinase A catalytic subunit (PKAc) lead to cortisol-producing adenomas and the debilitating stress disorder adrenal Cushing's syndrome. This article reviews classical studies regarding PKA localization in glucocorticoid-producing adrenal cells and synthesizes recent evidence of disrupted PKA localization and selective regulatory interactions in adrenal pathology.

Single-cell spatial transcriptomics reveals molecular patterns of selective neuronal vulnerability to α-synuclein pathology in a transgenic mouse model of Lewy body disease.

One of the unifying pathological hallmarks of Parkinson's disease (PD) and dementia with Lewy bodies (DLB) is the presence of misfolded, aggregated, and often phosphorylated forms of the protein α-synuclein in neurons. α-Synuclein pathology appears in select populations of neurons throughout various cortical and subcortical regions, and little is currently known about why some neurons develop pathology while others are spared. Here, we utilized subcellular-resolution imaging-based spatial transcriptomics (IST) in a transgenic mouse model that overexpresses wild-type human α-synuclein (α-syn-tg) to evaluate patterns of selective neuronal vulnerability to α-synuclein pathology. By performing post-IST immunofluorescence for α-synuclein phosphorylated at Ser129 (pSyn), we identified cell types in the cortex and hippocampus that were vulnerable or resistant to developing pSyn pathology. Next, we investigated the transcriptional underpinnings of the observed selective vulnerability using a set of custom probes to detect genes involved in α-synuclein processing and toxicity. We identified expression of the kinase:substrate pair Plk2 , which phosphorylates α-synuclein at Ser129, and human SNCA ( hSNCA ), as underlying the selective vulnerability to pSyn pathology. Finally, we performed differential gene expression analysis, comparing non-transgenic cells to pSyn - and pSyn + α-syn-tg cells to reveal gene expression changes downstream of hSNCA overexpression and pSyn pathology, which included pSyn-dependent alterations in mitochondrial and endolysosomal genes. This study provides a comprehensive use case of IST, yielding new biological insights into the formation of α-synuclein pathology and its downstream effects in a PD/DLB mouse model.

Modeling the distribution of enzymes on lipid vesicles: A novel framework for surface-mediated reactions in coagulation

Blood coagulation is a network of biochemical reactions wherein dozens of proteins act collectively to initiate a rapid clotting response. Coagulation reactions are lipid-surface dependent, and this dependence is thought to help localize coagulation to the site of injury and enhance the association between reactants. Current mathematical models of coagulation either do not consider lipid as a variable or do not agree with experiments where lipid concentrations were varied. Since there is no analytic rate law that depends on lipid, only apparent rate constants can be derived from enzyme kinetic experiments. We developed a new mathematical framework for modeling enzymes reactions in the presence of lipid vesicles. Here the concentrations are such that only a fraction of the vesicles harbor bound enzymes and the rest remain empty. We call the lipid vesicles with and without enzyme TF:VIIa+ and TF:VIIa− lipid, respectively. Since substrate binds to both TF:VIIa+ and TF:VIIa− lipid, our model shows that excess empty lipid acts as a strong sink for substrate. We used our framework to derive an analytic rate equation and performed constrained optimization to estimate a single, global set of intrinsic rates for the enzyme–substrate pair. Results agree with experiments and reveal a critical lipid concentration where the conversion rate of the substrate is maximized, a phenomenon known as the template effect. Next, we included product inhibition of the enzyme and derived the corresponding rate equations, which enables kinetic studies of more complex reactions. Our combined experimental and mathematical study provides a general framework for uncovering the mechanisms by which lipid mediated reactions impact coagulation processes.

Reversing the relative time courses of the peptide bond reaction with oligopeptides of different lengths and charged amino acid distributions in the ribosome exit tunnel

The kinetics of the protein elongation cycle by the ribosome depends on intertwined factors. One of these factors is the electrostatic interaction of the nascent protein with the ribosome exit tunnel. In this computational biology theoretical study, we focus on the rate of the peptide bond formation and its dependence on the ribosome exit tunnel electrostatic potential profile. We quantitatively predict how oligopeptides of variable lengths can affect the peptide bond formation rate. We applied the Michaelis-Menten model as previously extended to incorporate the mechano-biochemical effects of forces on the rate of reaction at the catalytic site of the ribosome. For a given pair of carboxy-terminal amino acid substrate at the P- and an aminoacyl-tRNA at the A-sites, the relative time courses of the peptide bond formation reaction can be reversed depending on the oligopeptide sequence embedded in the tunnel and their variable lengths from the P-site. The reversal is predicted to occur from a shift in positions of charged amino acids upstream in the oligopeptidyl-tRNA at the P-site. The position shift must be adjusted by clever design of the oligopeptide probes using the electrostatic potential profile along the exit tunnel axial path. These predicted quantitative results bring strong evidence of the importance and relative contribution of the electrostatic interaction of the ribosome exit tunnel with the nascent peptide chain during elongation.

(NHC)Ni(II)-catalyzed regioselective hydroalkenylation of norbornene derivatives: fine-tuning of NHC ligands and donor alkenes.

This study introduced a novel cross-hydroalkenylation approach for addressing a resilient challenge in C5-exo-selective norbornene derivative functionalization. The process, guided by mutual interactions among the NHC-Ni catalyst and the substrate pairs, ensured highly chemo- and regio-selective insertion.

IDPpub: Illuminating the Dark Phosphoproteome Through PubMed Mining

Global phosphoproteomics experiments quantify tens of thousands of phosphorylation sites. However, data interpretation is hampered by our limited knowledge on functions, biological contexts, or precipitating enzymes of the phosphosites. This study establishes a repository of phosphosites with associated evidence in biomedical abstracts, using deep learning-based natural language processing techniques. Our model for illuminating the dark phosphoproteome through PubMed mining (IDPpub) was generated by fine-tuning BioBERT, a deep learning tool for biomedical text mining. Trained using sentences containing protein substrates and phosphorylation site positions from 3000 abstracts, the IDPpub model was then used to extract phosphorylation sites from all MEDLINE abstracts. The extracted proteins were normalized to gene symbols using the National Center for Biotechnology Information gene query, and sites were mapped to human UniProt sequences using ProtMapper and mouse UniProt sequences by direct match. Precision and recall were calculated using 150 curated abstracts, and utility was assessed by analyzing the CPTAC (Clinical Proteomics Tumor Analysis Consortium) pan-cancer phosphoproteomics datasets and the PhosphoSitePlus database. Using 10-fold cross validation, pairs of correct substrates and phosphosite positions were extracted with an average precision of 0.93 and recall of 0.94. After entity normalization and site mapping to human reference sequences, an independent validation achieved a precision of 0.91 and recall of 0.77. The IDPpub repository contains 18,458 unique human phosphorylation sites with evidence sentences from 58,227 abstracts and 5918 mouse sites in 14,610 abstracts. This included evidence sentences for 1803 sites identified in CPTAC studies that are not covered by manually curated functional information in PhosphoSitePlus. Evaluation results demonstrate the potential of IDPpub as an effective biomedical text mining tool for collecting phosphosites. Moreover, the repository (http://idppub.ptmax.org), which can be automatically updated, can serve as a powerful complement to existing resources.

One-dimensional hydrodynamic model for the isothermal-evaporation of an axisymmetric-unbounded liquid bridge

In this manuscript, we examine the isothermal-evaporation of an unbounded axisymmetric liquid bridge confined between parallel-planar similar or chemically different substrates using both theory and experiments for axial Bond numbers 0.04 and 0.16. A perturbation analysis of the governing equations of motion produced a one-dimensional equation model, and this model made it possible to compute the interface profile evolution. Glass-acrylic or acrylic-acrylic substrate pairs (top-bottom) were used in experiments, with purified water as the liquid. Transient liquid-bridge volume estimates and contact angle data were extracted via image analysis at fixed time intervals and compared with results from the profiles that were computed using the one-dimensional equation model. There was good agreement when comparing the volume estimates and contact angle data.

Mechanistic Static Model based Prediction of Transporter Substrate Drug-Drug Interactions Utilizing Atorvastatin and Rifampicin.

An in vitro relative activity factor (RAF) technique combined with mechanistic static modeling was examined to predict drug-drug interaction (DDI) magnitude and analyze contributions of different clearance pathways in complex DDIs involving transporter substrates. Atorvastatin and rifampicin were used as a model substrate and inhibitor pair. In vitro studies were conducted with transfected HEK293 cells, hepatocytes and human liver microsomes. Prediction success was defined as predictions being within twofold of observations. The RAF method successfully translated atorvastatin uptake from transfected cells to hepatocytes, demonstrating its ability to quantify transporter contributions to uptake. Successful translation of atorvastatin's in vivo intrinsic hepatic clearance (CLint,h,in vivo) from hepatocytes to liver was only achieved through consideration of albumin facilitated uptake or through application of empirical scaling factors to transporter-mediated clearances. Transporter protein expression differences between hepatocytes and liver did not affect CLint,h,in vivo predictions. By integrating cis and trans inhibition of OATP1B1/OATP1B3, atorvastatin-rifampicin (single dose) DDI magnitude could be accurately predicted (predictions within 0.77-1.0 fold of observations). Simulations indicated that concurrent inhibition of both OATP1B1 and OATP1B3 caused approximately 80% of atorvastatin exposure increases (AUCR) in the presence of rifampicin. Inhibiting biliary elimination, hepatic metabolism, OATP2B1, NTCP, and basolateral efflux are predicted to have minimal to no effect on AUCR. This study demonstrates the effective application of a RAF-based translation method combined with mechanistic static modeling for transporter substrate DDI predictions and subsequent mechanistic interpretation.

StereoPHIP: Stereoselective Parahydrogen-Induced Polarization.

Parahydrogen-induced polarization (PHIP) followed by polarization transfer to 13 C is a rapidly developing technique for the generation of 13 C-hyperpolarized substrates. Chirality plays an essential role in living systems and differential metabolism of enantiomeric pairs of metabolic substrates is well documented. Inspired by asymmetric hydrogenation, here we report stereoPHIP, which involves the addition of parahydrogen to a prochiral substrate with a chiral catalyst followed by polarization transfer to 13 C spins. We demonstrate that parahydrogen could be rapidly added to the prochiral precursor to both enantiomers of lactic acid (D and L), with both the (R,R) and (S,S) enantiomers of a chiral rhodium(I) catalyst to afford highly 13 C-hyperpolarized (over 20 %) L- and D-lactate ester derivatives, respectively, with excellent stereoselectivity. We also show that the hyperpolarized 1 H signal decays obtained with the (R,R) and (S,S) catalysts were markedly different. StereoPHIP expands the scope of conventional PHIP to the production of 13 C hyperpolarized chiral substrates with high stereoselectivity.

StereoPHIP: Stereoselective Parahydrogen‐Induced Polarization

AbstractParahydrogen‐induced polarization (PHIP) followed by polarization transfer to 13C is a rapidly developing technique for the generation of 13C‐hyperpolarized substrates. Chirality plays an essential role in living systems and differential metabolism of enantiomeric pairs of metabolic substrates is well documented. Inspired by asymmetric hydrogenation, here we report stereoPHIP, which involves the addition of parahydrogen to a prochiral substrate with a chiral catalyst followed by polarization transfer to 13C spins. We demonstrate that parahydrogen could be rapidly added to the prochiral precursor to both enantiomers of lactic acid (D and L), with both the (R,R) and (S,S) enantiomers of a chiral rhodium(I) catalyst to afford highly 13C‐hyperpolarized (over 20 %) L‐ and D‐lactate ester derivatives, respectively, with excellent stereoselectivity. We also show that the hyperpolarized 1H signal decays obtained with the (R,R) and (S,S) catalysts were markedly different. StereoPHIP expands the scope of conventional PHIP to the production of 13C hyperpolarized chiral substrates with high stereoselectivity.

Iridium(III)-Catalyzed Intermolecular C(sp3 )-H Amidation for the Synthesis of Chiral 1,2-Diamines.

Chiral 1,2-diamines are privileged scaffolds among bioactive natural products, active pharmaceutical ingredients, ligands for transition-metal-based asymmetric catalysis and organocatalysts. Despite this interest, the construction of chiral 1,2-diamine motifs still remains a challenge. To address this, an iridium(III)-catalyzed intermolecular C(sp3 )-H amidation reaction was developed. This method relies on the design of a new, cheap and cleavable exo-protecting/directing group derived from camphorsulfonic acid, which is directly installed from easily accessible precursors, and furnishes scalemic free 1,2-diamines upon cleavage of both nitrogen substituents. It was found applicable to both α-secondary and α-tertiary-1,2-diamines, for which a two-step protocol involving intermolecular olefin hydroamination and C(sp3 )-H amidation was developed. Kinetic and computational studies provided insights into the observed reactivity difference between pairs of diastereoisomeric substrates.

Iridium(III)‐Catalyzed Intermolecular C(sp3)‐H Amidation for the Synthesis of Chiral 1,2‐Diamines

AbstractChiral 1,2‐diamines are privileged scaffolds among bioactive natural products, active pharmaceutical ingredients, ligands for transition‐metal‐based asymmetric catalysis and organocatalysts. Despite this interest, the construction of chiral 1,2‐diamine motifs still remains a challenge. To address this, an iridium(III)‐catalyzed intermolecular C(sp3)‐H amidation reaction was developed. This method relies on the design of a new, cheap and cleavable exo‐protecting/directing group derived from camphorsulfonic acid, which is directly installed from easily accessible precursors, and furnishes scalemic free 1,2‐diamines upon cleavage of both nitrogen substituents. It was found applicable to both α‐secondary and α‐tertiary‐1,2‐diamines, for which a two‐step protocol involving intermolecular olefin hydroamination and C(sp3)‐H amidation was developed. Kinetic and computational studies provided insights into the observed reactivity difference between pairs of diastereoisomeric substrates.

Engineering of substrate specificity in a plant cell-wall modifying enzyme through alterations of carboxyl-terminal amino acid residues.

Structural determinants of substrate recognition remain inadequately defined in broad specific cell-wall modifying enzymes, termed xyloglucan xyloglucosyl transferases (XETs). Here, we investigate the Tropaeolum majus seed TmXET6.3 isoform, a member of the GH16_20 subfamily of the GH16 network. This enzyme recognises xyloglucan (XG)-derived donors and acceptors, and a wide spectrum of other chiefly saccharide substrates, although it lacks the activity with homogalacturonan (pectin) fragments. We focus on defining the functionality of carboxyl-terminal residues in TmXET6.3, which extend acceptor binding regions in the GH16_20 subfamily but are absent in the related GH16_21 subfamily. Site-directed mutagenesis using double to quintuple mutants in the carboxyl-terminal region - substitutions emulated on barley XETs recognising the XG/penta-galacturonide acceptor substrate pair - demonstrated that this activity could be gained in TmXET6.3. We demonstrate the roles of semi-conserved Arg238 and Lys237 residues, introducing a net positive charge in the carboxyl-terminal region (which complements a negative charge of the acidic penta-galacturonide) for the transfer of xyloglucan fragments. Experimental data, supported by molecular modelling of TmXET6.3 with the XG oligosaccharide donor and penta-galacturonide acceptor substrates, indicated that they could be accommodated in the active site. Our findings support the conclusion on the significance of positively charged residues at the carboxyl terminus of TmXET6.3 and suggest that a broad specificity could be engineered via modifications of an acceptor binding site. The definition of substrate specificity in XETs should prove invaluable for defining the structure, dynamics, and function of plant cell walls, and their metabolism; these data could be applicable in various biotechnologies.

The Architecture of Metabolic Networks Constrains the Evolution of Microbial Resource Hierarchies.

Microbial strategies for resource use are an essential determinant of their fitness in complex habitats. When facing environments with multiple nutrients, microbes often use them sequentially according to a preference hierarchy, resulting in well-known patterns of diauxic growth. In theory, the evolutionary diversification of metabolic hierarchies could represent a mechanism supporting coexistence and biodiversity by enabling temporal segregation of niches. Despite this ecologically critical role, the extent to which substrate preference hierarchies can evolve and diversify remains largely unexplored. Here, we used genome-scale metabolic modeling to systematically explore the evolution of metabolic hierarchies across a vast space of metabolic network genotypes. We find that only a limited number of metabolic hierarchies can readily evolve, corresponding to the most commonly observed hierarchies in genome-derived models. We further show how the evolution of novel hierarchies is constrained by the architecture of central metabolism, which determines both the propensity to change ranks between pairs of substrates and the effect of specific reactions on hierarchy evolution. Our analysis sheds light on the genetic and mechanistic determinants of microbial metabolic hierarchies, opening new research avenues to understand their evolution, evolvability, and ecology.

Asymmetric droplet evaporation on inclined surfaces

A computational model based on disjoining pressure is developed to study the asymmetric evaporation of droplets on an inclined substrate. In the proposed formulation, the disjoining pressure is combined with the arbitrary Lagrange-Euler method (ALE) to describe the solid-liquid interaction and investigate the contact angle hysteresis during evaporation. This model can be used to simulate the droplet evaporation modes of constant contact radius (CCR), constant contact angle (CCA) and “Stick-Slip” (mix evaporation mode), while the results are in good agreement with the experimental data. The obtained droplet shape for different inclined angles shows an asymmetric variation in contact angle at the front and rear ends, with the mean contact angle decreasing as the tilt angle increases. The evaporation flux is symmetric in the case of a horizontal substrate, however, when the substrate is tilted, the front and rear contact angles are modified, which leads to a larger evaporation flux on the rear contact line. Moreover, the asymmetric droplet morphology leads to asymmetric temperature field and vortex in the initial stage of evaporation; as evaporation proceeds, however, the flow and temperature fields within the droplet exhibit a gradual convergence towards symmetry. Moreover, when droplets evaporate on a pair of substrates with complementary tilt angles, their lifetimes exhibit a remarkable similarity. From the evaporation rate evolution, it yields that the larger the percentage of pinning time on the contact line, the faster the evaporation rate of the droplet. The evaporation rate of droplets was 14.76% higher at a tilt angle of α=0° than the case of α=45°.

27‐4: Development of a novel glass‐cell FAIMS olfactory sensor and measurements of complex odors

High‐Field Asymmetric Waveform Ion Mobility Spectrometry (FAIMS) utilizes the characteristics of nonlinear ion mobility at high electric fields to filter ions. We propose a novel glass‐cell FAIMS olfactory sensor that mimics the olfactory sense of living organisms by applying LCD mass production process technology. This olfactory sensor has a simple configuration consisting of an odor molecule ionization section and a glass‐cell FAIMS sensor, which is a pair of glass substrates with patterned electrodes. The ionized odor molecules pass through the glass‐cell FAIMS sensor, and the filtered ion molecules are detected. We have experimented with different odorous components, including fermented beverages (wine) and animal excrement (cat urine, stool), to create unique “odor fingerprints”.

Comparison of Monophosphine and Bisphosphine Precatalysts for Ni-Catalyzed Suzuki-Miyaura Cross-Coupling: Understanding the Role of the Ligation State in Catalysis.

Practical advances in Ni-catalyzed Suzuki-Miyaura cross-coupling (SMC) have been limited by a lack of mechanistic understanding of phosphine ligand effects. While bisphosphines are commonly used in these methodologies, we have observed instances where monophosphines can provide comparable or higher levels of reactivity. Seeking to understand the role of ligation state in catalysis, we performed a head-to-head comparison study of C(sp2)-C(sp2) Ni SMCs catalyzed by mono and bisphosphine precatalysts using six distinct substrate pairings. Significant variation in optimal precatalyst was observed, with the monophosphine precatalyst tending to outperform the bisphosphines with electronically deactivated and sterically hindered substrates. Mechanistic experiments revealed a role for monoligated (P1Ni) species in accelerating the fundamental organometallic steps of the catalytic cycle, while highlighting the need for bisligated (P2Ni) species to avoid off-cycle reactivity and catalyst poisoning by heterocyclic motifs. These findings provide guidelines for ligand selection against challenging substrates and future ligand design tailored to the mechanistic demands of Ni-catalyzed SMCs.

(NHC)Ni(II)-Directed Insertions and Higher Substituted Olefin Synthesis from Simple Olefins

ConspectusWell-controlled olefin insertion is critical for achieving catalytic and productive bulk and fine-chemical synthesis. Developing efficient and selective methods for meeting diverse insertion demands is extremely noteworthy, as it supports numerous transformations. The challenges are related to improving catalyst performance and selectivity control and uniting previously unreactive substrate pairs to achieve higher molecular structural complexity and utility. Nickel catalysts have received persistent attention in higher substituted olefin synthesis and polymerization, and numerous new strategies have been established to fulfill the ever-changing demands. This Account focuses on the recent progress based on N-heterocyclic carbene (NHC) ligands and nickel catalysts in our laboratory in using simple terminal olefins as olefin donors or acceptors.It begins with a brief history of olefin codimerization and the major advances in hydrovinylation achieved by other research groups using ethylene as an olefin donor. It then describes problems related to the reductive elimination that can occur when both the hydrometalated alkene and NHC are on the catalyst. It emphasizes the impact of NHC catalyst generation methods on the competing reactivity. Next, it explains the principal challenges and great opportunities in using our method (with α-olefins as olefin donors and alkenyl sources) to replace intermolecular reductive hydroalkenylation reactions (which require rare and more expensive alkenyl halides and boronic acids as reactants, alongside a stoichiometric amount of metallic reagents). The Account then illustrates the potential uses of our method for solving challenging organic synthesis problems using tailor-made (NHC)Ni(II) catalysts to allow redox-neutral catalytic cycles based on high chemo- and regioselective cross-insertion controls. It shows that upon optimal steric and electronic cooperation between the NHC, olefin donor, and olefin acceptor, regiodivergent insertion and convergent synthesis can be achieved easily.In the course of our work, we uncovered several unique insights into regulating (anti-)Markovnikov hydronickelation, carbonickelation, hydrocarbonation, ring closure, 1,3-allyl shift, isomerization, and catalyst regeneration under green, neutral, and mild-temperature conditions. These insights are also outlined here, along with theoretical calculations that offer additional understandings of the insertion reactivity and selectivity differences observed between the NHC and the highly related phosphorus-based Ni(II) hydride-catalyzed cross-hydroalkenylation and cycloisomerization systems.Compared to traditional olefin and cyclic structure synthesis technology, such as olefin cross-metathesis, enyne cyclization, and cross-coupling reactions, the new catalyst systems often offer previously inaccessible product structural characteristics, substrate scope, and outcomes. In particular, the method is effective for the catalytic synthesis of unsymmetrical and functionalized 1,1-disubstituted olefins (a.k.a. gem-olefins), 1,4-dienes (a.k.a. skipped dienes), conjugated dienes, endo- and exocyclic olefins, fused and spiro rings, and aromatic products. These syntheses are variously achieved by cross-hydroalkenylation, insertion-induced rearrangement, cycloadditions, and other approaches inspired by our investigations and detailed in this Account. Cross-hydroalkenylation can be achieved with high enantioselectivity by application of carefully designed and structurally flexible C1 and C2 chiral NHC ligands, yielding a pool of chiral branched alkenes and 1,4-dienes directly from simple chemical feedstocks used in industry. This Account will draw further attention to green alkenylation and the related development of redox-neutral catalytic cycles.

Homocysteine impedes neurite outgrowth recovery after intracerebral haemorrhage by downregulating pCAMK2A

Hyperhomocysteinemia (HHcy) is independently associated with poorer long-term prognosis in patients with intracerebral haemorrhage (ICH); however, the effect and mechanisms of HHcy on ICH are still unclear. Here, we evaluated...