Drosophila HCN mediates gustatory homeostasis by preserving sensillar transepithelial potential in sweet environments.

Establishing transepithelial ion disparities is crucial for sensory functions in animals. In insect sensory organs called sensilla, a transepithelial potential, known as the sensillum potential (SP), arises through active ion transport across accessory cells, sensitizing receptor neurons such as mechanoreceptors and chemoreceptors. Because multiple receptor neurons are often co-housed in a sensillum and share SP, niche-prevalent overstimulation of single sensory neurons can compromise neighboring receptors by depleting SP. However, how such potential depletion is prevented to maintain sensory homeostasis remains unknown. Here, we find that the Ih-encoded hyperpolarization-activated cyclic nucleotide-gated (HCN) channel bolsters the activity of bitter-sensing gustatory receptor neurons (bGRNs), albeit acting in sweet-sensing GRNs (sGRNs). For this task, HCN maintains SP despite prolonged sGRN stimulation induced by the diet mimicking their sweet feeding niche, such as overripe fruit. We present evidence that Ih-dependent demarcation of sGRN excitability is implemented to throttle SP consumption, which may have facilitated adaptation to a sweetness-dominated environment. Thus, HCN expressed in sGRNs serves as a key component of a simple yet versatile peripheral coding that regulates bitterness for optimal food intake in two contrasting ways: sweet-resilient preservation of bitter aversion and the previously reported sweet-dependent suppression of bitter taste.

White matter tract density index is associated with disability in multiple sclerosis.

BACKGROUND: The association between common neuroradiological markers of multiple sclerosis (MS) and clinical disability is weak. Given that the disability in patients with MS may depend on the underlying structural connectivity of the brain, our study aimed to examine the association between white matter tracts affected by MS and the patients' disability using a new tract density index (TDI). METHOD: This study included 53 patients diagnosed with MS, examined between 2019 and 2020. Manual lesion segmentation was performed on fluid-attenuated inversion recovery (FLAIR) images, and the density of white matter tracts encompassing the lesion (i.e., TDI) was calculated. Correlation analysis was employed to assess the association between TDI and disability. Additionally, the relationship between disability, TDI, and lesion-derived network metrics was examined by computing a partial correlation network. RESULTS: The TDI significantly correlated with the expanded disability status scale (EDSS) (r = 0.30, p = 0.03). Furthermore, the patient's disability is linked solely through TDI to lesion-derived network metrics -a key metric that 'bridges' the gap between the brain lesion and disability. CONCLUSIONS: In this study, MS lesions encompassing regions with high white matter tract density were associated and linked with severe physical disability. These findings indicate that TDI may be an outcome predictor that may connect radiologic findings to clinical practice.

Optimizing Contact Between Older Adults and Senior Services: South Korean users' Perceptions of Gateway Institutions.

In response to the growing need for effective policy implementation strategies for older adults in South Korea, we propose the establishment of a user-centered institution tightly integrated with policies, termed, "comprehensive gateway institution for older adults." This research addresses the challenges 231 older adult users face when navigating various health policies. Our survey findings revealed that these users experienced difficulties in dealing with the current policy approach for older adults. The respondents expressed their expectations for improved service access through the proposed gateway institution. They emphasized the importance of universal access to services and the need for personalized offerings that consider their unique circumstances, physical abilities, and skills. By incorporating these research outcomes into practice, we can help lay the groundwork for more effective policies measures and create a system that better meets the needs of older adults in the future.

Effects of nature on restorative and cognitive benefits in indoor environment.

This study investigates the effects of natural exposure in an indoor environment on restorative quality and cognitive ability. Thirty participants were shown nature at three different indoor sites: baseline, indoor (some vegetation), and semi-indoor (a large amount of vegetation and view to sky) for five minutes. After viewing, they completed an assessment of restoration and a cognitive task, and their electroencephalography (EEG) was recorded. Compared to the baseline, the sites with nature resulted in restorative (higher perceived restoration scores) and cognitive (higher working memory performance and lower delta-to-theta ratio (DTR), delta-to-alpha ratio (DAR), theta-to-beta ratio (TBR), and alpha-to-beta ratio (ABR) responses) benefits. These findings further our understanding of the effects of exposure to nature on restorative and cognitive benefits in an indoor environment, and help to build guidance for future research on the effects of nature indoors and designing restorative- and cognitive-enhancing indoor spaces.

Validation of Ultrasound and Computed Tomography-Based Risk Stratification System and Biopsy Criteria for Cervical Lymph Nodes in Preoperative Patients With Thyroid Cancer.

OBJECTIVE: This study aimed to validate the risk stratification system (RSS) and biopsy criteria for cervical lymph nodes (LNs) proposed by the Korean Society of Thyroid Radiology (KSThR). MATERIALS AND METHODS: This retrospective study included a consecutive series of preoperative patients with thyroid cancer who underwent LN biopsy, ultrasound (US), and computed tomography (CT) between December 2006 and June 2015. LNs were categorized as probably benign, indeterminate, or suspicious according to the current US- and CT-based RSS and the size thresholds for cervical LN biopsy as suggested by the KSThR. The diagnostic performance and unnecessary biopsy rates were calculated. RESULTS: A total of 277 LNs (53.1% metastatic) in 228 patients (mean age ± standard deviation, 47.4 years ± 14) were analyzed. In US, the malignancy risks were significantly different among the three categories (all P < 0.001); however, CT-detected probably benign and indeterminate LNs showed similarly low malignancy risks (P = 0.468). The combined US + CT criteria stratified the malignancy risks among the three categories (all P < 0.001) and reduced the proportion of indeterminate LNs (from 20.6% to 14.4%) and the malignancy risk in the indeterminate LNs (from 31.6% to 12.5%) compared with US alone. In all image-based classifications, nodal size did not affect the malignancy risks (short diameter [SD] ≤ 5 mm LNs vs. SD > 5 mm LNs, P ≥ 0.177). The criteria covering only suspicious LNs showed higher specificity and lower unnecessary biopsy rates than the current criteria, while maintaining sensitivity in all imaging modalities. CONCLUSION: Integrative evaluation of US and CT helps in reducing the proportion of indeterminate LNs and the malignancy risk among them. Nodal size did not affect the malignancy risk of LNs, and the addition of indeterminate LNs to biopsy candidates did not have an advantage in detecting LN metastases in all imaging modalities.

Effect of alanine versus serine at position 88 of human transthyretin mutants on the protein stability.

Human transthyretin (TTR) is a homo-tetrameric plasma protein associated with a high percentage of ß-sheet forming amyloid fibrils. It accumulates in tissues or extracellular matrices to cause amyloid diseases. Free energy simulations with thermodynamic integration based on all-atom molecular dynamics simulations have been carried out to analyze the effects of the His88 â†’ Ala and Ser mutations on the stability of human TTR. The calculated free energy change differences (ΔΔG) caused by the His88 â†’ Ala and His88 â†’ Ser mutations are -1.84 ± 0.86 and 7.56 ± 0.55 kcal/mol, respectively, which are in excellent agreement with prior reported experimental values. The simulation results show that the H88A mutant is more stable than the wild type, whereas the H88S mutant is less stable than the wild type. The free energy component analysis shows that the contribution to the free energy change difference (ΔΔG) for the His88 â†’ Ala and His88 â†’ Ser mutations mainly arise from electrostatic and van der Waals interactions, respectively. The electrostatic term stabilizes the H88A mutant more than the wild type, but the van der Waals interaction destabilizes the H88S mutant relative to the wild type. Individual residue contributions to the free energy change show neighboring residues exert stabilizing and destabilizing influence on the mutants. The implications of the simulation results for understanding the stabilizing and destabilizing effect and its contribution to protein stability are discussed.

Enlarged perivascular spaces are associated with decreased brain tau deposition.

AIMS: The aim of this study was to investigate the associations of enlarged perivascular spaces (EPVS) in the basal ganglia (BG) and centrum semiovale (CSO) with beta-amyloid (Aß) and tau deposition in older adults with a diverse cognitive spectrum. METHODS: A total of 163 (68 cognitively normal and 95 cognitively impaired) older participants underwent [11 C] Pittsburgh compound B and [18 F] AV-1451 PET, and MRI. EPVS in the BG and CSO and other small vessel disease markers, such as white matter hyperintensities, lacunes, and deep and lobar microbleeds, were assessed. RESULTS: Increased EPVS in the BG showed a significant association with lower cerebral tau deposition, even after controlling for other small vessel disease markers. Further exploratory analyses showed that this association was significant in cognitively impaired, Aß-positive, or APOE4-positive individuals, but not significant in the cognitively normal, Aß-negative, or APOE4-negative participants. In contrast to EPVS in the BG, EPVS in the CSO did not have any relationship with cerebral tau deposition. In addition, none of the two types of EPVS were associated with cerebral Aß deposition. CONCLUSION: Brain tau deposition appears to be reduced with increased EPVS in the BG, especially in individuals with cognitive impairment, pathological amyloid burden, or genetic Alzheimer's disease risk.

Free energy simulations to study mutational effect of a conserved residue, Trp24, on stability of human serum retinol-binding protein.

Human serum retinol-binding protein (RBP) is a plasma transport protein for vitamin A. RBP is a prime subclass of lipocalins, which bind nonpolar ligands within a ß-barrel. To understand the role of Trp 24, one of the highly conserved residues in RBP, free energy simulations have been carried out to understand the effects of the mutations from Trp at position 24 to Leu, Phe, and Tyr in the apo-RBP on its thermal stability. We examine various unfolded systems to study the dependence of the free energy differences on the denatured structure. Our calculated free energy difference values for the three mutations are in excellent agreement with the experimental values when the initial coordinates of the seven-residue peptide segments truncated from the crystal structure are used for the denatured systems. Our free energy change differences for the Trp→Leu, Trp→Phe, and Trp→Tyr mutations are 2.50 ± 0.69, 2.58 ± 0.50, and 2.49 ± 0.48 kcal/mol, respectively, when the native-like seven-residue peptides are used as models for the denatured systems. The main contributions to the free energy change differences for the Trp24→Leu and Trp24→Phe mutations are mainly from van der Waals and covalent interactions, respectively. Electrostatic, van der Waals and covalent terms equally contribute to the free energy change difference for the Trp24→Tyr mutation. The free energy simulation helps understand the detailed microscopic mechanism of the stability of the RBP mutants relative to the wild type and the role of the highly conserved residue, Trp24, of the human RBP.Communicated by Ramaswamy H. Sarma.

Simulation analysis of selective alanine mutation effect on stability of human prion protein.

Prion diseases are neurodegenerative disorders caused by spongiform degeneration of the brain. Understanding the fundamental mechanism of prion protein aggregation caused by mutations is very crucial to resolve the pathology of prion diseases. To help understand the roles of individual residues on the stability of the human prion protein, the computational method of free energy simulations based on atomistic molecular dynamics trajectories is applied to Phe175 → Ala, Val180 → Ala, and Val209 → Ala mutations of the human prion protein. The simulations show that all three alanine mutations destabilize the human prion protein. The calculated free energy change differences, ΔΔG, for the Phe175 → Ala, Val180 → Ala, and Val209 → Ala mutations are in good agreement with the experimental values. The significant destabilizing effects on the mutants relative to the wild-type protein arise from van der Waals terms. Furthermore, our free energy decomposition analysis shows that the major contribution to destabilizing the V180A and V209A mutants relative to the wild-type protein is originated from van der Waals interactions from residues near the mutation sites. In contrast, the contribution to destabilizing the F175A mutant is mainly caused by van der Waals interactions from residues near and far away from the mutation site. Our results show that the free energy simulation with a thermodynamic integration approach for selected alanine scanning mutations is beneficial for understanding the detailed mechanism of human prion protein destabilization, specific residues' role, and the hydrophobic effect on protein stability.Communicated by Ramaswamy H. Sarma.

In Vitro Demonstration of Human Lipoyl Synthase Catalytic Activity in the Presence of NFU1.

Lipoyl synthase (LS) catalyzes the last step in the biosynthesis of the lipoyl cofactor, which is the attachment of sulfur atoms at C6 and C8 of an n-octanoyllysyl side chain of a lipoyl carrier protein (LCP). The protein is a member of the radical S-adenosylmethionine (SAM) superfamily of enzymes, which use SAM as a precursor to a 5'-deoxyadenosyl 5'-radical (5'-dA·). The role of the 5'-dA· in the LS reaction is to abstract hydrogen atoms from C6 and C8 of the octanoyl moiety of the substrate to initiate subsequent sulfur attachment. All radical SAM enzymes have at least one [4Fe-4S] cluster that is used in the reductive cleavage of SAM to generate the 5'-dA·; however, LSs contain an additional auxiliary [4Fe-4S] cluster from which sulfur atoms are extracted during turnover, leading to degradation of the cluster. Therefore, these enzymes catalyze only 1 turnover in the absence of a system that restores the auxiliary cluster. In Escherichia coli, the auxiliary cluster of LS can be regenerated by the iron-sulfur (Fe-S) cluster carrier protein NfuA as fast as catalysis takes place, and less efficiently by IscU. NFU1 is the human ortholog of E. coli NfuA and has been shown to interact directly with human LS (i.e., LIAS) in yeast two-hybrid analyses. Herein, we show that NFU1 and LIAS form a tight complex in vitro and that NFU1 can efficiently restore the auxiliary cluster of LIAS during turnover. We also show that BOLA3, previously identified as being critical in the biosynthesis of the lipoyl cofactor in humans and Saccharomyces cerevisiae, has no direct effect on Fe-S cluster transfer from NFU1 or GLRX5 to LIAS. Further, we show that ISCA1 and ISCA2 can enhance LIAS turnover, but only slightly.

[Association between Cerebral Small Vessel and Alzheimer's Disease]. / ì•Œì¸ í•˜ì´ë¨¸ë³‘ê³¼ 뇌소혈관질환의 연관성.

Cerebral small vessel disease (CSVD) includes vascular lesions detected on brain MRI, such as white matter hyperintensities, lacunar infarctions, microbleeds, or enlarged perivascular spaces. There is accumulating evidence that vascular changes may play an important role in development of Alzheimer's disease (AD), and CSVD lesions detected on brain MRI were reported to be associated with ß-amyloid and tau proteins accumulation. As the vascular contribution has therapeutic potential, it is important to understand the association of CSVD with AD and AD biomarkers. This review begins with a brief introduction of AD and AD biomarkers, explains the association between AD and vascular changes, and then details the pathogenesis and MR imaging findings of CSVD. Afterwards, we discuss the association of CSVD with AD and AD biomarkers.

Models and Molecular Markers of Spermatogonial Stem Cells in Vertebrates: To Find Models in Nonmammals.

Spermatogonial stem cells (SSCs) are the germline stem cells that are essential for the maintenance of spermatogenesis in the testis. However, it has not been sufficiently understood in amphibians, reptiles, and fish because numerous studies have been focused mainly on mammals. The aim of this review is to discuss scientific ways to elucidate SSC models of nonmammals in the context of the evolution of testicular organization since rodent SSC models. To further understand the SSC models in nonmammals, we point out common markers of an SSC pool (undifferentiated spermatogonia) in various types of testes where the kinetics of the SSC pool appears. This review includes the knowledge of (1) common molecular markers of vertebrate type A spermatogonia including putative SSC markers, (2) localization of the markers on the spermatogonia that have been reported in previous studies, (3) highlighting the most common markers in vertebrates, and (4) suggesting ways of finding SSC models in nonmammals.

Validation of Ultrasound Risk Stratification Systems for Cervical Lymph Node Metastasis in Patients with Thyroid Cancer.

A malignancy risk stratification system (RSS) for cervical lymph nodes (LNs) has not been fully established. This study aimed to validate the current RSS for the diagnosis of cervical LN metastasis in thyroid cancer. In total, 346 LNs from 282 consecutive patients between December 2006 and June 2015 were included. We determined the malignancy risk of each ultrasound (US) feature and performed univariable and multivariable logistic regression analyses. Each risk category from the Korean Society of Thyroid Radiology (KSThR) and the European Thyroid Association (ETA) was applied to calculate malignancy risks. The effects of size, number of suspicious features, and primary tumor characteristics were analyzed to refine the current RSS. Suspicious features including echogenic foci, cystic change, hyperechogenicity, and abnormal vascularity were independently predictive of malignancy (p ≤ 0.045). The malignancy risks of probably benign, indeterminate, and suspicious categories were 2.2-2.5%, 26.8-29.0%, and 85.8-87.4%, respectively, according to the KSThR and ETA criteria. According to the ETA criteria, 15.1% of LNs were unclassifiable. In indeterminate LNs, multiplicity of the primary tumor was significantly associated with malignancy (odds ratio, 6.53; p = 0.004). We refined the KSThR system and proposed a US RSS for LNs in patients with thyroid cancer.

Significant Therapeutic Effects of Adult Human Neural Stem Cells for Spinal Cord Injury Are Mediated by Monocyte Chemoattractant Protein-1 (MCP-1).

The limited capability of regeneration in the human central nervous system leads to severe and permanent disabilities following spinal cord injury (SCI) while patients suffer from no viable treatment option. Adult human neural stem cells (ahNSCs) are unique cells derived from the adult human brain, which have the essential characteristics of NSCs. The objective of this study was to characterize the therapeutic effects of ahNSCs isolated from the temporal lobes of focal cortical dysplasia type IIIa for SCI and to elucidate their treatment mechanisms. Results showed that the recovery of motor functions was significantly improved in groups transplanted with ahNSCs, where, in damaged regions of spinal cords, the numbers of both spread and regenerated nerve fibers were observed to be higher than the vehicle group. In addition, the distance between neuronal nuclei in damaged spinal cord tissue was significantly closer in treatment groups than the vehicle group. Based on an immunohistochemistry analysis, those neuroprotective effects of ahNSCs in SCI were found to be mediated by inhibiting apoptosis of spinal cord neurons. Moreover, the analysis of the conditioned medium (CM) of ahNSCs revealed that such neuroprotective effects were mediated by paracrine effects with various types of cytokines released from ahNSCs, where monocyte chemoattractant protein-1 (MCP-1, also known as CCL2) was identified as a key paracrine mediator. These results of ahNSCs could be utilized further in the preclinical and clinical development of effective and safe cell therapeutics for SCI, with no available therapeutic options at present.

Magnetoactive Acoustic Topological Transistors.

Topological field-effect transistor is a revolutionary concept that physical fields are used to switch on and off quantum topological states of the condensed matter. Although this emerging concept has been explored in electronics, how to realize it in the acoustic realm remains elusive. In this work, a class of magnetoactive acoustic topological transistors capable of on-demand switching on and off topological states and reconfiguring topological edges with external magnetic fields is presented. The key mechanism is to harness magnetic fields to tune air-cavity volumes within acoustic chambers, thus breaking or preserving the inversion symmetry to manifest or conceal the quantum valley Hall effect. To switch the topological transport beyond the in-plane routes, a magneto-tuned non-topological band gap to allow or forbid the wave transport out-of-plane is harnessed. With the reversible magnetic control, on-demand switching of topological routes to realize topological field-effect waveguides and wave regulators is demonstrated. Analogous to the impact of semiconductor transistors on modern electronics, this work may expand the scope of topological acoustics by achieving unprecedented functions in acoustic modulation.

Modulation of human transthyretin stability by the mutations at histidine 88 studied by free energy simulation.

Human transthyretin (TTR) is a homotetrameric plasma protein associated with a high percentage of ß-sheet, which forms amyloid fibrils and accumulates in tissues or extracellular matrix to cause amyloid diseases. Free energy simulations based on all-atom molecular dynamics simulations were carried out to analyze the effects of the His88 → Arg, Phe, and Tyr mutations on the stability of human TTR. The calculated free energy change differences (ΔΔG) caused by the His → Arg, Phe, and Tyr mutations at position 88 are 6.48 ± 0.45, -9.99 ± 0.54, and 2.66 ± 0.33 kcal/mol, respectively. These calculated free energy change differences between wild type and the mutants are in excellent agreement with prior experimental values. Our simulation results show that the wild type of the TTR is more stable than H88R and H88Y mutants, whereas it is less stable than the H88F mutant. The free energy component analysis shows that the primary contribution to the free energy change difference (ΔΔG) for the His → Arg mutation arises from electrostatic interaction; the ΔΔG for the His → Phe mutation is from van der Waals and electrostatic interactions and that for the His → Tyr mutation from covalent interaction. The simulation results show that the free energy calculation with thermodynamic integration is beneficial for understanding the detailed microscopic mechanism of protein stability. The implications of the results for understanding stabilizing and destabilizing effect of the mutation and the contribution to protein stability are discussed.

Constructive adaptation of 3D-printable polymers in response to typically destructive aquatic environments.

In response to environmental stressors, biological systems exhibit extraordinary adaptive capacity by turning destructive environmental stressors into constructive factors; however, the traditional engineering materials weaken and fail. Take the response of polymers to an aquatic environment as an example: Water molecules typically compromise the mechanical properties of the polymer network in the bulk and on the interface through swelling and lubrication, respectively. Here, we report a class of 3D-printable synthetic polymers that constructively strengthen their bulk and interfacial mechanical properties in response to the aquatic environment. The mechanism relies on a water-assisted additional cross-linking reaction in the polymer matrix and on the interface. As such, the typically destructive water can constructively enhance the polymer's bulk mechanical properties such as stiffness, tensile strength, and fracture toughness by factors of 746% to 790%, and the interfacial bonding by a factor of 1,000%. We show that the invented polymers can be used for soft robotics that self-strengthen matrix and self-heal cracks after training in water and water-healable packaging materials for flexible electronics. This work opens the door for the design of synthetic materials to imitate the constructive adaptation of biological systems in response to environmental stressors, for applications such as artificial muscles, soft robotics, and flexible electronics.

Unassisted N-acetyl-phenylalanine-amide transport across membrane with varying lipid size and composition: kinetic measurements and atomistic molecular dynamics simulation.

Biological membranes are essential to preserve structural integrity and regulate functional properties through the permeability of nutrients, pharmaceutical drugs, and neurotransmitters of a living cell. The movement of acetylated and amidated phenylalanine (NAFA) across 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membrane bilayers is investigated to probe physical transport. The rate of transport is measured experimentally applying parallel artificial membrane permeation assay (PAMPA). At the physiological temperature, 310 K, the measured time constants in the neutral pH were ∼6 h in DOPC and ∼3 h in POPC, while in a more acidic condition, at a pH 4.8, the time constants were ∼8 h in both lipids. Computationally, we have expanded our transport study of three aromatic dipeptides across a bilayer composed of DOPC18. In this study, we have examined the effects of lipid composition and bilayer size on the passive transport of NAFA by simulating the dipeptide in three different bilayers, with 50 DOPC lipids, 50 POPC lipids, and 40 POPC molecules. Specifically, atomistic molecular dynamics simulations with umbrella sampling were used to calculate the potential of mean force for the passive permeation of NAFA across the bilayers. Diffusion constants were then calculated by numerically solving the Smoluchowski equation. Permeability coefficients and mean first passage times were then calculated. Structural properties - Ramachandran plots, sidechain torsions, peptide insertion angles, radial distribution functions, and proximal peptide water molecules - were also examined to determine the effect of system size and lipid type. In terms of systems size, we observed a small decrease in the highest barrier of the potential of mean force and fewer sampled sidechain dihedral angle conformations with 40 versus 50 POPC lipids due to weaker membrane deformations within a smaller lipid bilayer. In terms of lipid type, DOPC contains two monounsaturated acyl chains compared to only one such acyl chain in POPC; therefore, DOPC bilayers are less ordered and more easily deformed, as seen by a much broader potential of mean force profile. The NAFA in DOPC lipid also transitioned to an internally hydrogen-bonded backbone conformation at lower membrane depths than in POPC. Similarly, as for other aromatic dipeptides, NAFA tends to insert into the membrane sidechain-first, remains mostly desolvated in the membrane center, and exhibits slow reorientations within the bilayer in both DOPC and POPC. With a joint experimental and computational study we have gained a new insight into the rate of transport and the underlying microscopic mechanism in different lipid bilayer conditions of the simplest hydrophobic aromatic dipeptide.Communicated by Ramaswamy H. Sarma.

Single-cell RNA sequencing of human nail unit defines RSPO4 onychofibroblasts and SPINK6 nail epithelium.

Research on human nail tissue has been limited by the restricted access to fresh specimen. Here, we studied transcriptome profiles of human nail units using polydactyly specimens. Single-cell RNAseq with 11,541 cells from 4 extra digits revealed nail-specific mesenchymal and epithelial cell populations, characterized by RSPO4 (major gene in congenital anonychia) and SPINK6, respectively. In situ RNA hybridization demonstrated the localization of RSPO4, MSX1 and WIF1 in onychofibroblasts suggesting the activation of WNT signaling. BMP-5 was also expressed in onychofibroblasts implicating the contribution of BMP signaling. SPINK6 expression distinguished the nail-specific keratinocytes from epidermal keratinocytes. RSPO4+ onychofibroblasts were distributed at close proximity with LGR6+ nail matrix, leading to WNT/ß-catenin activation. In addition, we demonstrated RSPO4 was overexpressed in the fibroblasts of onychomatricoma and LGR6 was highly expressed at the basal layer of the overlying epithelial component, suggesting that onychofibroblasts may play an important role in the pathogenesis of onychomatricoma.