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The zebrafish caudal fin has become a popular model to study cellular and molecular mechanisms of regeneration due to its high regenerative capacity, accessibility for experimental manipulations, and relatively simple anatomy. The formation of a regenerative epidermis and blastema are crucial initial events and tightly regulated. Both the regenerative epidermis and the blastema are highly organized structures containing distinct domains, and several signaling pathways regulate the formation and interaction of these domains. Bone is the major tissue regenerated from the progenitor cells of the blastema. Several cellular mechanisms can provide source cells for blastemal (pre-)osteoblasts, including dedifferentiation of differentiated osteoblasts and de novo formation from other cell types, providing intriguing examples of cellular plasticity. In recent years, omics analyses and single-cell approaches have elucidated genetic and epigenetic regulation, increasing our knowledge of the surprisingly complex coordination of various mechanisms to achieve successful restoration of a seemingly simple structure.

Keratin 17 (K17) is a multifaceted cytoskeletal protein that is not commonly expressed in the epidermis under normal physiological conditions. However, in psoriasis, K17 is overexpressed in the suprabasal layer of the epidermis and plays an important role in the pathogenesis of the disease. In this review, we have summarized our findings and those reported in other studies concerning the pathogenic functions of K17, as well as the mechanisms underlying the increase in K17 expression in psoriasis. K17 exerts both pro-proliferative and pro-inflammatory effects on keratinocytes. Moreover, K17 peptides trigger autoreactive T cells and promote psoriasis-related cytokine production. In turn, these cytokines modulate the expression, stability, and protein-protein interactions of K17 through transcriptional and translational regulation and post-translational modification of K17 in keratinocytes. Thus, a K17/T-cell/cytokine autoimmune loop is implicated in the pathogenesis of psoriasis, which is supported by the fact that therapies targeting K17 have achieved good outcomes in psoriasis-like mouse models. Future perspectives of K17 in psoriasis have also been discussed to provide potential directions for further studies.

Heavy metals like cadmium (Cd) are a serious threat for living organisms due to increased global environmental pollution. Aloe ferox Mill. is a very important species of the genus Aloe, especially from a medical point of view, because it is a source of biologically active substances and it is often used in the traditional medicine. Since the knowledge about the effect of heavy metals, like Cd on growth of Aloe species is scarce, this study showed the effect of Cd on anatomical, morphological and physiological properties of this important medicinal plant species. We found that Cd affected the leaf morphology and decreased biomass of the aboveground plant part. Cd also affected cuticle deposition and increased the number of stomata in the abaxial and adaxial epidermis of leaves. Roots treated with Cd showed lower lignification and decreased level of membrane lipids peroxidation than control roots. Cadmium concentration was the highest in the roots; however, relatively high portion of Cd is being translocated to the leaves. This might be dangerous as Aloe is well known medicinal plant with content of active secondary metabolites mainly in the leaves.

It is important to detect specific genes expressed in the guard cells, which control gas exchange and play key roles in response to drought and salt stresses. Due to the genetic transformation of Chinese cabbage (Brassica rapa) has not been well developed, in situ RT-PCR is a valuable option for detecting guard cell specific genes. We reported an optimized protocol of in situ RT-PCR by using an FAMA homologous gene Bra001929 in Brassica rapa. FAMA in Arabidopsis has been verified to be specially expressed in guard cells. We designed specific RT-PCR primers and optimized the protocol in terms of the (a) reverse transcription time, (b) blocking time, (c) antigen-antibody incubation time, and (d) washing temperature. Our approach provides a sensitive and effective in situ RT-PCR method for locating expression in the guard cells in Brassica rapa. Moreover, we proved the guard cell specific expression of Bra001929 in the epidermis indicating its’ applicability as a marker gene for guard cells of Brassica rapa.

The application of medical devices to repair skin damage is clinically accepted and natural polymer enjoys an important role in this field, such as collagen or hyaluronic acid, etc. However, the biosafety and efficacy of these implants are still challenged. In this study, a skin damage animal model was prepared by UV-photoaging and recombinant humanized type III collagen (rhCol III) was applied as a bioactive material to implant in vivo to study its biological effect, comparing with saline and uncrosslinked hyaluronic acid (HA). Animal skin conditions were non-invasively and dynamically monitored during the 8 weeks experiment. Histological observation, specific gene expression and other molecular biological methods were applied by the end of the animal experiment. The results indicated that rhCol III could alleviate the skin photoaging caused by UV radiation, including reduce the thickening of epidermis and dermis, increase the secretion of Collagen I (Col I) and Collagen III (Col III) and remodel of extracellular matrix (ECM). Although the cell-material interaction and mechanism need more investigation, the effect of rhCol III on damaged skin was discussed from influence on cells, reconstruction of ECM, and stimulus of small biological molecules based on current results. In conclusion, our findings provided rigorous biosafety information of rhCol III and approved its potential in skin repair and regeneration. Although enormous efforts still need to be made to achieve successful translation from bench to clinic, the recombinant humanized collagen showed superiorities from both safety and efficacy aspects.

Background Prurigo nodularis (PN) is a debilitating, difficult-to-treat, intensely pruritic, chronic inflammatory skin disease characterized by hyperkeratotic skin nodules. The pathogenesis of PN is not well understood but is believed to involve cross talk between sensory nerve fibers, immune cells, and the epidermis. It is centered around the neuroimmune cytokine IL-31, driving an intractable itch–scratch cycle. Objective We sought to provide a comprehensive view of the transcriptomic changes in PN skin and characterize the mechanism of action of the anti–IL-31 receptor inhibitor nemolizumab. Method RNA sequencing of biopsy samples obtained from a cohort of patients treated with the anti–IL-31 receptor inhibitor nemolizumab and taken at baseline and week 12. Generation and integration of patient data with RNA-Seq data generated from reconstructed human epidermis stimulated with IL-31 and other proinflammatory cytokines. Results Our results demonstrate that nemolizumab effectively decreases IL-31 responses in PN skin, leading to effective suppression of downstream inflammatory responses including TH2/IL-13 and TH17/IL-17 responses. This is accompanied by decreased keratinocyte proliferation and normalization of epidermal differentiation and function. Furthermore, our results demonstrate how transcriptomic changes associated with nemolizumab treatment correlate with improvement in lesions, pruritus, stabilization of extracellular matrix remodeling, and processes associated with cutaneous nerve function. Conclusion These data demonstrate a broad response to IL-31 receptor inhibition with nemolizumab and confirm the critical upstream role of IL-31 in PN pathogenesis.

Dermal fibroblasts are strategically positioned underneath the basal epidermis layer to support keratinocyte proliferation and extracellular matrix production. In inflammatory conditions, these fibroblasts produce cytokines and chemokines that promote the chemoattraction of immune cells into the dermis and the hyperplasia of the epidermis, two characteristic hallmarks of psoriasis. However, how dermal fibroblasts specifically contribute to psoriasis development remains largely uncharacterized. In this study, we investigated through which cytokines and signaling pathways dermal fibroblasts contribute to the inflammatory features of psoriatic skin. We show that dermal fibroblasts from lesional psoriatic skin are important producers of inflammatory mediators, including IL-6, CXCL8, and CXCL2. This increased cytokine production was found to be regulated by ZFP36 family members ZFP36, ZFP36L1, and ZFP36L2, RNA-binding proteins with mRNA-degrading properties. In addition, the expression of ZFP36 family proteins was found to be reduced in chronic inflammatory conditions that mimic psoriatic lesional skin. Collectively, these results indicate that dermal fibroblasts are important producers of cytokines in psoriatic skin and that reduced expression of ZFP36 members in psoriasis dermal fibroblasts contributes to their inflammatory phenotype.

Gall-inducing organisms change the development of their host plant organs, resulting in ontogenetic patterns not observed in the non-galled plants. Distinct taxa induce galls on Schinus spp., manipulating meristematic patterns in the host plant in distinct ways. Here we report ontogenetic novelties induced in the lateral buds of S. engleri by Eucecidoses minutanus, a Cecidosidae, whose galls have been poorly understood.The anatomy, histochemistry, and histometry of galls in distinct phases of development, non-galled buds, and stems of Schinus engleri were analyzed in parallel with the instars of E. minutanus to detail the morphogenetic changes in the host with each larval stage.Ontogenetic phases of the galls were intricately associated with larval development. First and second-instar larvae induced pericycle and pith cells to dedifferentiate into the gall inner meristem, where hyperplasia and cell hypertrophy characterized the growth and development phase of the gall. The innermost layers were lipid-rich nutritive cells that lined the larval chamber. Additional vascular bundle rows were produced in young galls. Third and fourth instar-larvae were associated with the gall maturation phase: centripetal lignification of the outer parenchyma cell layers, epidermal stratification, and activation of a cambium-like meristem (CLM). The CLM activity resulted in new layers of nutritive cells that differentiated inward as the first layers of nutritive cells were consumed by E. minutanus larvae, and, also, in more parenchyma cell layers that formed outward. All tissues between the innermost layer of nutritive tissue that surround the gall chamber and the outermost layer of the dermal system that externally covers the gall form the gall wall, and increased in thickness until the end of gall maturation.E. minutanus induces a structurally complex globoid stem gall, modifying all host plant tissues and stimulating a novel meristematic pattern in S. engleri. The gall developmental stages are each related to specific gall-inducing instars, as gall development progresses according to the development of E. minutanus.

The main objective of this study was to investigate the effects of bacterial cellulose hydrogel (BCH) incorporated into montmorillonite (MMT) and its underlying mechanisms of action on a skin wound healing mouse model following pressure injury model. Komagataeibacter hansenii was used to obtain 5 cm in diameter and 0.8 mm of thickness circular bacterial cellulose (BC) sheets, which were incorporated with MMT by deposition ex-site using a 0.1% MMT suspension (100 rpm for 24 h at 28 °C). Afterward, Fourier Transform Infrared Spectroscopy (FT-IR) and Scanning Electron Microscopy (SEM) were used to characterize the bacterial cellulose hydrogel incorporated into montmorillonite (BCH-MMT). The pressure injury model was assessed by macroscopic and histological analysis in male Swiss mice. Both, BC and BCH-MMT, showed a typical FTIR spectrum of cellulosic substrates with pronounces bands around 3344, 2920, 1637, and 1041 cm−1 while microparticles of MMT dispersed uniformly throughout BC were revealed by SEM photographs. Animals treated with BCH-MMT showed significant healing of pressure ulcers as demonstrated by reduced area of redness and spontaneous hyperalgesia, lower amounts of in-site inflammatory cells (to the same level as the positive control Dersani®) and ultimately, complete epidermis re-epithelialization and tissue regeneration. Altogether, these findings suggest that a modified BCH-MMT film could serve as scaffolding for skin tissue engineering and potentially as a novel dressing material for pressure injury.

Molting is critical for growth, development, and survival in insects. As the main components of cuticle, dynamic change of chitin is consistent with molting process. Chitinase is main enzyme to mediate chitin metabolism in the old cuticle. To avoid over-degrading chitin from the new cuticle, expression of chitinase must be precisely regulated. In this study, we performed microRNA-sequencing to investigate expression change of microRNAs in silkworm epidermis during molting process. Comparative microRNA transcriptomic analysis from different molting stages and 20-hydroxyecdysone (20E) treatment identified bmo-miR-282-5p as candidate. By bioinformatic analysis, chitinase 5 (BmCht5) was predicted to be a target of bmo-miR-282-5p. Meanwhile, temporal expression analysis revealed that BmCht5 only expressed at molting D3 stage, whereas bmo-miR-282-5p showed converse pattern, in which its transcript signal disappeared at this time point. Furthermore, luciferase assay and agomir treatment demonstrated that bmo-miR-282-5p suppressed transcript of BmCht5 in vivo. As a result, injection of 282-5p agomir triggered 40% death due to molting failure. In addition, RNA interference (RNAi)-mediated silencing of BmCht5 caused 30% developmental defect. Taken together, our data demonstrates the coordinated regulation of chitinase 5 by conserved miR-282-5p and 20E signaling pathway is essential for the normal molting process in the domesticated silkworm. This article is protected by copyright. All rights reserved.

Staphylococcus aureus is a skin commensal microorganism commonly colonizing healthy humans. Nevertheless, S. aureus can also be responsible for cutaneous infections and contribute to flare-up of inflammatory skin diseases such as atopic dermatitis (AD), which is characterized by dysbiosis of the skin microbiota with S. aureus as the predominant species. However, the role of major virulence factors of this pathogen such as phenol-soluble modulin (PSM) toxins in epidermal inflammation remains poorly understood. Stimulation of primary human keratinocytes with sublytic concentrations of synthetic and purified PSM α3 resulted in upregulation of a large panel of pro-inflammatory chemokine and cytokine gene expression, including CXCL1, CXCL2, CXCL3, CXCL5, CXCL8, CCL20, IL-1α, IL-1β, IL-6, IL-36γ and TNF-α, while inducing the release of CXCL8, CCL20, TNF-α and IL-6. In addition, using S. aureus culture supernatant from mutants deleted from genes encoding either α-type PSMs or all PSM production, PSMs were shown to be the main factors of S. aureus secretome responsible for pro-inflammatory mediator induction in human keratinocytes. On the other hand, α-type PSM-containing supernatant triggered an intense induction of pro-inflammatory mediator expression and secretion during both topical and basal layer stimulation of an ex vivo model of human skin explants, a physiologically relevant model of pluristratified epidermis. Taken together, the results of this study show that PSMs and more specifically α-type PSMs are major virulence factors of S. aureus inducing a potent inflammatory response during infection of the human epidermis and could thereby contribute to AD flare-up through exacerbation of skin inflammation.

Introduction The skin is comprised of various kinds of cells and has three layers, the epidermis, dermis and subcutaneous adipose tissue. Stem cells in each tissue duplicate themselves and differentiate to supply new cells that function in the tissue, and thereby maintain the tissue homeostasis. In contrast, senescent cells accumulate with age and secrete senescence-associated secretory phenotype (SASP) factors that impair surrounding cells and tissues, which lowers the capacity to maintain homeostasis in each tissue. Previously, we found Gremlin 2 (GREM2) as a novel SASP factor in the skin and reported that GREM2 suppressed the differentiation of adipose-derived stromal/stem cells. In the present study, we investigated the effects of GREM2 on stem cells in the epidermis and dermis. Methods To examine whether GREM2 expression and the differentiation levels in the epidermis and dermis are correlated, the expressions of GREM2, stem cell markers, an epidermal differentiation marker Keratin 10 (KRT10) and a dermal differentiation marker type 3 procollagen were examined in the skin samples (n = 14) randomly chosen from the elderly where GREM2 expression level is high and the individual differences of its expression are prominent. Next, to test whether GREM2 affects the differentiation of skin stem cells, cells from two established lines (an epidermal and a dermal stem/progenitor cell model) were cultured and induced to differentiate, and recombinant GREM2 protein was added. Results In the human skin, the expression levels of GREM2 varied among individuals both in the epidermis and dermis. The expression level of GREM2 was not correlated with the number of stem cells, but negatively correlated with those of both an epidermal and a dermal differentiation markers. The expression levels of epidermal differentiation markers were significantly suppressed by the addition of GREM2 in the three-dimensional (3D) epidermis generated with an epidermal stem/progenitor cell model. In addition, by differentiation induction, the expressions of dermal differentiation markers were induced in cells from a dermal stem/progenitor cell model, and the addition of GREM2 significantly suppressed the expressions of the dermal differentiation markers. Conclusions GREM2 expression level did not affect the numbers of stem cells in the epidermis and dermis but affects the differentiation and maturation levels of the tissues, and GREM2 suppressed the differentiation of stem/progenitor cells in vitro. These findings suggest that GREM2 may contribute to the age-related reduction in the capacity to maintain skin homeostasis by suppressing the differentiation of epidermal and dermal stem/progenitor cells., Highlights • In epidermis, the expression levels of GREM2 and KRT10 were negative correlation. • In dermis, the expression levels of GREM2 and Pro-COL3 were negative correlation. • GREM2 suppressed epidermal stem/progenitor cell differentiation in vitro. • GREM2 suppressed dermal stem/progenitor cell differentiation in vitro.

Introduction Artificial dermis is an effective therapeutic method for full-thickness dermal defects. However, the currently available artificial dermis made of porcine or bovine type I collagen has several limitations such as incomplete epithelialization and delayed migration of fibrogenic and angiogenic cells into the graft. We previously developed a composite dermal graft containing a mixture of moon jellyfish collagen and porcine type I collagen, and reported its stimulatory effect on both the re-epithelialization of the epidermis and the migration of fibrogenic and angiogenic cells into the graft. In the present study, we examined whether the same effect was observed by administering jellyfish collagen solution externally onto an artificial dermal graft made of bovine type I collagen. Methods We used a 6 mm full-thickness wound defect model. Moon jellyfish collagen was prepared as a concentrated 0.5% solution and dripped externally onto a transplanted artificial dermal graft made of bovine type I collagen. Wound repair and long-term dermal tissue remodeling were compared between mice administered jellyfish collagen solution on the bovine collagen graft and those transplanted with a composite dermal graft containing the same amounts of jellyfish and bovine collagens. The stimulatory effect of jellyfish collagen solution was also evaluated using diabetic dB/dB mice. Results External administration of jellyfish collagen solution onto the bovine collagen graft significantly accelerated wound closure compared to control saline. It also decreased the number of inflammatory cells infiltrating the wound and suppressed absorption of the transplanted graft, as well as reduced subsequent scar formation. Furthermore, external administration of jellyfish collagen solution onto the bovine collagen graft improved the delayed wound healing in diabetic model mice, and this effect was superior to that of the currently used basic fibroblast growth factor. Conclusions External administration of moon jellyfish collagen solution onto a bovine collagen graft significantly accelerated physiological wound healing and prevented excessive scar formation. It also improved wound closure in diabetic model mice, confirming its therapeutic application for intractable skin ulcers caused by impaired wound healing., Highlights • Impaired wound healing is frequently observed in elderly and diabetic patients. • Moon jellyfish collagen accelerates wound closure after full-thickness dermal defect in mice. • Jellyfish collagen used as an external medicine stimulates the elongation of regenerating epithelial cells. • Externally applied jellyfish collagen improves wound healing in diabetic model mice.

Abroma augusta L. known as Devil’s cotton belongs to Malvaceae. The exploratory study aimed to study the morphological and anatomical characteristics of the aerial parts of A. augusta L. from Semarang. The transverse section of the aerial parts was made by a simple method (fresh preparation) and then observed under a binocular microscope with an optilab. All characteristics were observed and then compared with the references. The collected data were analyzed descriptively and quantitatively. In summary, the results showed that A. augusta L. was an evergreen shrub (small tree) with orthotropic and plagiotropic branches and polymorphous leaves. The inflorenscence was found in the terminal and axillar plagiotropic branching with bisex, actinomorphic, and pentamerous flowers. The fruit was unique (obconical capsule with a rounded base and truncate-tip with 5 angled wings) including cotton fibers and numerous black seeds. The petiole was composed of epidermis, collenchyma, cortical parenchyma, sclerenchyma, vascular bundle, mucilaginous ducts, and pith. The dorsiventral leaf was composed of upper and lower epidermis, palisade, and spongy parenchyma. The stomata type was ranunculaceous (anomocytic) while the guard cell was kidney-shaped. The stomata density on the abaxial leaf was higher than that of the adaxial leaf. The stellate and unicellular non-glandular trichomes, and capitate glandular trichomes were found abundantly on the petiole and leaf blade. These morphological and anatomical studies are important to support the identification as a part of the conservation effort of the plant. Further studies are recommended to investigate the root morphology and anatomy and also biochemical characteristics of each part of the plant in order to obtain complete plant identification.

In this study, the toxic effects of paraquat, one of the most commercially sold herbicides in the world, and the protective role of green tea leaf extract (GTLE) against these effects were investigated. Allium cepa L. bulbs (n = 16) were used as test material. One hundred milligrams per liter dose of paraquat and 190 and 380 mg/L doses of GTLE were preferred. Paraquat toxicity was investigated with the help of physiological (percent germination, root length, and weight gain), cytogenetic (mitotic index = MI, micronucleus = MN, and chromosomal damages = CAs), biochemical (superoxide dismutase = SOD, catalase = CAT, malondialdehyde = MDA), and anatomical (meristematic cell damages) parameters. A. cepa bulbs were divided into 6 groups as 1 control and 5 applications. The control group was germinated with tap water, and the application groups were germinated with paraquat and two different doses of GTLE. Germination was carried out at room temperature for 72 h. At the end of the period, A. cepa bulbs were prepared for physiological, cytogenetic, biochemical, and anatomical analyzes using routine preparation techniques. As a result, paraquat application caused a decrease in physiological parameters and an increase in cytogenetic (except MI) and biochemical parameters. Compared to the control (group I), the germination percentage decreased by 38%, root length 12.5 times, and weight gain 5 times decreased in group IV treated with paraquat. MDA level increased 2.58 times, SOD activity 2.48 times, and CAT activity 4.51 times increased. Paraquat application caused a decrease in the percentage of MI and an increase in the number of MN and CAs. Paraquat application caused CAs in the form of fragment, sticky chromosome, unequal distribution of chromatin, bridge, nucleus with vacuoles, nucleus bud, and reverse polarization. In the meristematic cells of the root tips applied paraquat, unclearly vascular tissue, flattened cell nucleus, epidermis, and cortex cell deformation were observed. The application of GTLE together with paraquat caused an increase in the physiological parameter values and a decrease in the cytogenetic (except MI) and biochemical parameter values. An improvement in the severity of damages induced by paraquat was also observed in root tip meristematic cells. It was determined that the improvements observed in all these parameters were related to the dose of GTLE applied. The 380 mg/L dose of GTLE provided more protection than the 190 mg/L dose. Compared to group IV in which paraquat was applied, the germination percentage increased by 21%, root length 5.83 times, and weight gain 2.92 times increased in group VI administered 380 mg/L dose of GTLE. In addition, MDA level decreased 1.78 times, SOD activity 1.59 times and CAT activity 1.65 times. In conclusion, paraquat administration at a dose of 100 mg/L caused physiological, cytogenetic, biochemical, and anatomical toxicity in A. cepa bulbs. GTLE application, on the other hand, resulted in improvements in the severity of this toxicity induced by paraquat, depending on the dose. Therefore, GTLE can be used as an effective nutritional supplement to reduce or prevent the toxicity caused by environmental agents such as pesticides.

In the present study, the surface and external structure of petals, sepals, hypanthium and pedicel of eight Rosa species were examined. Micromorphological investigation of distribution and dimension of different trichome types, which were found in analysed plant parts was done using scanning electron microscopy (SEM). Our results showed that the adaxial epidermal cells of all analysed petals formed papillae, whereas the abaxial epidermis was with flat walls and had cuticle striation. Two types of trichomes, the capitate glandular and nonglandular trichomes (ribbon and acicles) were observed. Ribbon trichomes were presented on the adaxial side of all analysed sepals with the exception of R. spinosissima samples. Multicellular glandular trichomes were distributed on the surface of the hypanthium and the sepals of R. gallica, R. micrantha, R. pendulina, R. tomentosa and R. villosa samples. In addition, they were presented on the pedicel of most analysed species, except in R. dumetorum samples. Micromorphological analysis had shown that capitate glandular trichomes, based on their size, can be classified into two typeslong and short trichomes. Their presence, type and length were specific for each analysed species. Additionally, acicles were found only on the pedicel and hypanthium of R. spinosissima and R. gallica samples. This study provided useful information of different trichomes of genus Rosa which could be helpful in more accurate plant determination.

Some plants have been reported to have the water-induced shape memory effect (SME). Since the tissue system of most plant leaves is a biophysical sandwich-like structure composed of epidermis, mesophyll and veins, the leaves are actually bio-composites. In this paper, we experimentally investigated the contribution of different tissues to the SME in dried plant leaves (namely, soursop, sugar apple, and longan) and skeleton (i.e., cuticle and lamina removed) leaves (soursop only). Shape recovery in the leave strips, and skeleton soursop leave strips, all programmed (deformed) via rolling and drying, was activated by room temperature water, hot (90 °C) water, or pure heating (up to 230 °C) to uncover the influence of temperature in the SME. Although heating alone does not induce any apparent shape recovery, hot water causes much faster recovery than room temperature water. Besides, same as the influence of the maximum programming strain on the heating-responsive shape recovery in shape memory polymers, less programmed leaves (i.e., with smaller maximum programming strain) tend to recover faster. Together with other tests (including drying, wetting, drying/wetting, cyclic differential scanning calorimetry, etc.), the contributions/roles of different tissues on the phenomena observed in this study are revealed. This work is supported by the National Natural Science Foundation of China (Grant No. 11828201 and No. 11872207) and Natural Science Foundation of Jiangsu Province (Grant No. BK20200413).

Keratinocytes are the primary cell type in the epidermis and play an essential role in protecting the body against external insults. To investigate the modulatory potential of Korean red ginseng (KRG) on keratinocytes, we explored the possible mechanism underlying the promotion of keratinocyte migration. We determined the levels of TERT, HAS-2, SPTLC2, and CerS3 genes in HaCaT cells using qRT-PCR. In addition, changes in the levels of involucrin, filaggrin, and loricrin were measured using qRT-PCR and immunoblotting. Lastly, the keratinocyte migration promoting potential of KRG was determined using a cell migration assay and immunoblotting. KRG (100 and 200 μg/mL) treatment upregulated the telomerase reverse transcriptase gene independent of the UVB insult. KRG treatment upregulated the levels of HAS-2, SPTLC2, CerS3, and loricrin genes involved in the skin barrier function of keratinocytes. KRG promotes keratinocyte migration via NF-κB activation. KRG showed a direct modulatory effect on keratinocytes, and these modulatory functions may benefit skin health when KRG is used as a cosmeceutical or nutraceutical ingredient.

SummaryBackground: Hair follicles are produced in a cyclical manner and the machinery involved in the reproduction of these follicles is present since the fetal stage. Although extensive research has been done on the human hair follicle, very little is known about the importance of adhesion molecules in its development. Methods: We analyzed here, the immunoexpression of beta-1 integrin, p-cadherin, e- cadherin, and beta-catenin in hair follicles from 26 formalin-fixed and paraffin-embedded skin samples from human embryos and fetus between 12-23 weeks of gestational age. Findings: The adhesion molecules beta-1 integrin and e-cadherin/p-cadherin were expressed from 12 weeks and seemed to play a role in regulating epidermis invagination. Beta-catenin immuno stainingwas negative in all cases; down regulation of this protein may be necessary for fetal hair development and thus facilitating hair follicle down growth. Conclusion: Adhesion molecules are essential for hair follicle down growth and proliferation; integrins and cadherins play a major role in this process. More studies are needed to describe hair follicle development.

Erythrokeratodermia variabilis et progressiva (EKVP) is a rare genodermatosis of clinical and genetic heterogeneity, characterized by the manifestations of localized or disseminated persistent hyperkeratotic plagues and stationary to migratory transient erythematous patches. The majority of EKVP cases display an autosomal dominant mode of inheritance with incomplete penetrance, although recessive transmission has also been described. Mutations associated with EKVP have been primarily detected in connexin (Cx) genes. We herein reported a Chinese sporadic case of late-onset EKVP with a novel heterozygous missense mutation c.109G>A (p.V37M) in GJB4 (Cx30.3) gene, which resulted in a significant reduction of GJB4 expression in the epidermis of the patient. In accordance, while wild-type GJB4 localized at the cell membrane of HeLa cells forming intercellular junctions and intracellular puncta, V37M mutant variant was diffusely expressed within HeLa cells at a considerably lower level. Our findings reveal an essential role of GJB4 in the pathogenesis of EKVP and provides insights into the therapeutic potential of the disease.

The Ca(2+)-sensing receptor (CaSR) drives essential Ca(2+) and E-cadherin-mediated processes in the epidermis, including differentiation (Komuves et al., 2002), cell-to-cell adhesion (Tu et al., 2008) and epidermal barrier homeostasis in cells and young adult mice (Tunggal et al., 2005; Tu et al., 2012; Tu et al., 2019). We now report that decreased CaSR expression leads to impaired Ca(2+) signal propagation in aged (>22 months) mouse epidermis and (>79 years donor age) human keratinocytes. Baseline cytosolic Ca(2+) concentrations were higher, and capacitive Ca(2+) entry was lower in aged vs. young keratinocytes, even though endoplasmic reticulum (ER) stores were largely intact. As in CaSR knockout mice ((Epid)CaSR(−/−)), decreased CaSR expression led to decreased E-cadherin, PLC γ expression and a compensatory upregulation of Stromal Interaction Molecule 1 (STIM1). Pretreatment with the CaSR agonist NPS-R568 normalized Ca(2+) propagation and E-cadherin organization after experimental wounding. These results suggest that age-related defects in CaSR expression dysregulate normal keratinocyte and epidermal Ca(2+) signaling, leading to impaired E-cadherin expression, organization and function. These findings illustrate a innovative mechanism whereby Ca(2+) and E-cadherin-dependent functions are impaired in aging epidermis and suggest a new therapeutic approach by restoring CaSR function.

Background Acne is a common skin disorder that involves an infection inside the hair follicle, which is usually treated with antibiotics, resulting in unbalanced skin microbiota and microbial resistance. For this reason, we developed polymeric nanoparticles encapsulating thymol, a natural active compound with antimicrobial and antioxidant properties. In this work, optimization physicochemical characterization, biopharmaceutical behavior and therapeutic efficacy of this novel nanostructured system were assessed. Results Thymol NPs (TH-NP) resulted on suitable average particle size below 200 nm with a surface charge around − 28 mV and high encapsulation efficiency (80%). TH-NP released TH in a sustained manner and provide a slow-rate penetration into the hair follicle, being highly retained inside the skin. TH-NP possess a potent antimicrobial activity against Cutibacterium acnes and minor effect towards Staphylococcus epidermis, the major resident of the healthy skin microbiota. Additionally, the stability and sterility of developed NPs were maintained along storage. Conclusion TH-NP showed a promising and efficient alternative for the treatment of skin acne infection, avoiding antibiotic administration, reducing side effects, and preventing microbial drug resistance, without altering the healthy skin microbiota. Additionally, TH-NP enhanced TH antioxidant activity, constituting a natural, preservative-free, approach for acne treatment. Graphical Abstract

Mimosa pigra L. (Fabaceae), commonly known as giant mimosa, is one of the major invasive species, constituting a serious threat to agricultural crops and other plants in the biodiversity. With this study, we aimed to investigate the mechanism underlying the effects on plant growth and development upon exposure to two coumarins. Protein solutions were obtained from giant mimosa seedlings that had been exposed to the coumarins, and they were analyzed with the shotgun proteomic method by liquid chromatography–tandem mass spectrometry (LC–MS/MS). The proteins were identified in seedlings treated with coumarins. The control plants expressed 621 proteins, the plants treated with isoscopoletin expressed 613 proteins, and the plants treated with umbelliferone expressed 640 proteins. The classification of proteins by PANTHER software found that the coumarins interfered with proteins involved in cellular component organization or biogenesis and metabolic processes. STITCH analysis of protein expression after treatment with coumarins confirmed that CUL4, ESF, LNG2 and SCD2 played a role in plant growth and development. WAK1 and WAT1 were involved in cell wall and cell surface responses. At3g61030 and CIP111 were related to Ca2+ accumulation and CAND1, CUL4 and ML5 were linked with programmed cell death. It is worthwhile to mention that coumarins increased morphological changes in the epidermis, Ca2+ accumulation in the roots and protein expression in plant growth and development, morphological changes, Ca2+ accumulation as well as in programmed cell death.

Although various marine ingredients have been exploited for the development of cosmetic products, no previous study has examined the potential of seaweed extracellular vesicles (EV) in such applications. Our results revealed that EV from Codium fragile and Sargassum fusiforme effectively decreased α-MSH-mediated melanin synthesis in MNT-1 human melanoma cells, associated with downregulation of MITF (microphthalmia-associated transcription factor), tyrosinase and TRP1 (tyrosinase-related proteins 1). The most effective inhibitory concentrations of EV were 250 μg/ml for S. fusiforme and 25 μg/ml for C. fragile, without affecting the viability of MNT-1 cells. Both EV reduced melanin synthesis in the epidermal basal layer of a three-dimensional model of human epidermis. Moreover, the application of the prototype cream containing C. fragile EV (final 5 μg/ml) yielded 1.31% improvement in skin brightness in a clinical trial. Together, these results suggest that EV from C. fragile and S. fusiforme reduce melanin synthesis and may be potential therapeutic and/or supplementary whitening agents.

The loss of melanocytes in vitiligo is associated with architectural, transcriptional, and cellular perturbations of keratinocytes and manifests as a reduced proliferation potential in vitro and delayed re-epithelialization in vivo. To understand the molecular mechanisms underlying this delay, microRNA (miRNA) profiling was performed on split skin biopsies collected on Day 1 (basal level) and Day 14 (wound re-epithelialization) from nonlesional (NL) and lesional (L) skin of five subjects with stable nonsegmental vitiligo and 129 miRNAs were found to be differentially regulated between the NL and L healed epidermis. miR-21-5p, expressed at comparable levels on NL and L Day 1 samples, demonstrated significant upregulation during re-epithelialization. However, the extent of its upregulation was relatively lower in L (10 times compared to Day 1) as compared to NL skin (17 times compared to Day 1). The overexpression of miR-21 in keratinocytes led to a significant increase in the expression of proliferation markers (Ki67 and MCM6 messenger RNA, Ki67 positivity), along with an increase in keratinocyte migration. Using a small interfering RNA mediated knockdown approach, we further demonstrated that miR-21-5p mediates its effects by suppressing the expression of programmed cell death 4 (PDCD4) and mammary serine protease inhibitor (Maspin), both tumor-suppressor genes. Investigation of clinical samples corroborated the lower miR-21 levels and a higher expression of PDCD4 and Maspin in L Day 14 compared to the NL Day 14 epidermis. In conclusion, this study revealed that a relatively lower upregulation of miR-21-5p in L skin leads to significantly higher levels of PDCD4 and Maspin, delaying wound re-epithelialization by reducing the proliferation and migration of keratinocytes.

Human epidermis responds to ultraviolet (UV)B-induced damage by tolerating it, restoring it, or undergoing programmed cell death when the damage is massive. Recently, compounds rich in polyphenols, such as Vitis vinifera L. leaf extract (VVLe), have attracted a lot of interest for skin protection. We investigated the effect of VVLe pre-treatment (1 h) in a 2D model of HaCaT cells and in 3D organotypic cultures of normal human skin exposed to a single UVB dose to study the immediate specific events 1 h and the response orchestrated in the epidermal layer 24 h after irradiation, respectively. In both models, transmission electron microscopy analysis was carried out. The expression of the inducible keratin K17, the activation of both pSTAT3 and Nuclear Factor (NF)-κB signalling pathways, and the epidermal distribution of Toll-Like Receptor (TLR) 4 were assessed by immunofluorescence in the 2D and 3D model. In 3D organotypic cultures, thanks to the preservation of a multi-layered structure, the epidermal distribution of the differentiation biomarkers K10 and K14 as well as of K16 was analysed by immunofluorescence, while the release of interleukin (IL)-8 was evaluated by ELISA. In skin bioptic fragments, cytotoxicity and genotoxicity were investigated by LDH assay and Alkaline Comet assay, respectively, and then compared to cell proliferation. The epidermal distribution of the histone γ-H2AX, indicating the fragmented DNA, was analysed by immunofluorescence. In both experimental models, VVLe tuned UVB-induced K17 expression to a different extent in HaCaT cells and in the skin. In HaCaT cells, pSTAT3 activation was induced by UVB and reverted by VVLe pre-treatment. TLR4 expression was triggered by UVB in both models, but VVLe pre-treatment abolished this event only in HaCaT cells. NF-κB immunostaining increased both in the nucleus and in the cytoplasm only in HaCaT cells after UVB irradiation. In all irradiated skin samples, VVLe pre-treatment was not able to revert the inhibition of epidermal proliferation, K16 expression, and IL-8 secretion. The effectiveness of VVLe in contrasting the irradiation-induced genotoxicity still remains unclear. In conclusion, our study clearly shows that K17 is a robust marker induced in keratinocytes upon UVB stimulation and that this event can be reverted by a pre-treatment with VVLe. On the whole, these observations represent a novelty in the scenario of the complex relationships between the effects exerted by UVB rays on human skin and significantly improve the knowledge regarding the modulation of the early epidermal response induced by a single exposure to UVB in the presence of VVLe.

Background Epigallocatechin-3-gallate (EGCG) has been proven effective in treating viral warts. Since anticarcinogenic as well as anti-inflammatory properties are ascribed to the substance, its use has been evaluated in the context of different dermatoses. The effect of EGCG on interface dermatitis (ID), however, has not yet been explored. Objectives In this study, we investigated the effect of EGCG on an epidermal human in vitro model of ID. Methods Via immunohistochemistry, lesional skin of lichen planus patients and healthy skin were analysed concerning the intensity of interferon-associated mediators, CXCL10 and MxA. Epidermal equivalents were stained analogously upon ID-like stimulation and EGCG treatment. Monolayer keratinocytes were treated likewise and supernatants were analysed via ELISA while cells were processed for vitality assay or transcriptomic analysis. Results CXCL10 and MxA are strongly expressed in lichen planus lesions and induced in keratinocytes upon ID-like stimulation. EGCG reduces CXCL10 and MxA staining intensity in epidermis equivalents and CXCL10 secretion by keratinocytes upon stimulation. It furthermore minimizes the cytotoxic effect of the stimulus and downregulates a magnitude of typical pro-inflammatory cytokines that are crucial for the perpetuation of ID. Conclusions We provide evidence concerning anti-inflammatory effects of EGCG within a human in vitro model of ID. The capacity to suppress mediators that are centrally involved in disease perpetuation suggests EGCG as a potential topical therapeutic in lichen planus and other autoimmune skin diseases associated with ID.

Pigmented epidermotropic breast cancer metastases are a rarity, often clinically misdiagnosed as melanocytic lesions. Histopathologically, they show a dermal proliferation of neoplastic metastatic cells that extend to the overlying epidermis in a pattern identical to that seen in primary Paget disease (PD). Differential diagnosis should be established with entities with a similar presentation, such as pigmented mammary PD and malignant melanoma. Immunohistochemistry may be useful for this purpose. We present a new case of pigmented epidermotropic breast cancer metastases with a particularly unusual feature: the absence of dermal infiltration by neoplastic cells, thus considered as pure epidermotropic metastatic involvement.

As the outermost layer of plants, the epidermis serves as a critical interface between plants and the environment. During leaf development, the differentiation of specialized epidermal cell types, including stomatal guard cells, pavement cells, and trichomes, occurs simultaneously, each providing unique and pivotal functions for plant growth and survival. Decades of molecular-genetic and physiological studies have unraveled key players and hormone signaling specifying epidermal differentiation. However, most studies focus on only one cell type at a time, and how these distinct cell types coordinate as a unit is far from well-comprehended. Here we provide a review on the current knowledge of regulatory mechanisms underpinning the fate specification, differentiation, morphogenesis, and positioning of these specialized cell types. Emphasis is given to their shared developmental origins, fate flexibility, as well as cell cycle and hormonal controls. Furthermore, we discuss computational modeling approaches to integrate how mechanical properties of individual epidermal cell types and entire tissue/organ properties mutually influence each other. We hope to illuminate the underlying mechanisms coordinating the cell differentiation that ultimately generate a functional leaf epidermis.

We present a self-therapeutic nanoparticle for topical delivery to epidermal keratinocytes to prevent and treat psoriasis. Devoid of known chemical or biological antipsoriatic drugs, this sub-15 nm nanoparticle contains a 3 nm gold core and a shell of 1000 Da polyethylene glycol strands modified with 30% octadecyl chains. When it is applied to imiquimod-induced psoriasis mice without an excipient, the nanoparticle can cross the stratum corneum and preferentially enter keratinocytes. Applying the nanoparticles concurrently with imiquimod prevents psoriasis and downregulates genes that are enriched in the downstream of the interleukin-17 signaling pathway and linked to epidermis hyperproliferation and inflammation. Applying the nanoparticles after psoriasis is established treats the psoriatic skin as effectively as standard steroid and vitamin D analog-based therapy but without hair loss and skin wrinkling. The nanoparticles do not accumulate in major organs or induce long-term toxicity. Our nanoparticle offers a simple, safe, and effective alternative for treating psoriasis.

Oryza coarctata is the only wild rice species with significant salinity tolerance. The present work examines the role of the substantial rhizomatous tissues of O. coarctata in conferring salinity tolerance. Transition to an erect phenotype (shoot emergence) from prostrate growth of rhizome tissues is characterized by marked lignification and suberization of supporting sclerenchymatous tissue, epidermis, and bundle sheath cells in aerial shoot-proximal nodes and internodes in O. coarctata. With salinity, however, aerial shoot-proximal internodal tissues show reductions in lignification and suberization, most probably related to re-direction of carbon flux towards synthesis of the osmporotectant proline. Concurrent with hypolignification and reduced suberization, the aerial rhizomatous biomass of O. coarctata appears to have evolved mechanisms to store Na+ in these specific tissues under salinity. This was confirmed by histochemical staining, quantitative real-time reverse transcription–PCR expression patterns of genes involved in lignification/suberization, Na+ and K+ contents of internodal tissues, as well as non-invasive microelectrode ion flux measurements of NaCl-induced net Na+, K+, and H+ flux profiles of aerial nodes were determined. In O. coarctata, aerial proximal internodes appear to act as ‘traffic controllers’, sending required amounts of Na+ and K+ into developing leaves for osmotic adjustment and turgor-driven growth, while more deeply positioned internodes assume a Na+ buffering/storage role.

The relationship between photosynthesis, pigment accumulation, and the expression of key light-regulated genes in Solanum lycopersicum hp-1, hp-2 and hp-1.2 photomorphogenetic mutants under conditions of high-intensity light (2000 μm (photons) m−2s−1) was studied. The hp-2 mutant (LA3006) and the hp-1 mutants (LA4012 and LA3538) are deficient in DET1 (De-etiolated 1 and DDB1 (DNA DAMAGE-BINDING PROTEIN 1), respectively, which are components of the CDD complex (COP10, DDB1, DET1). HP mutants are superproducers of various pigments and are sensitive to light. We have shown that HIL (high-intensity light) causes a decrease in PSII activity after 24 and 72 h of irradiation, which was partially restored after 72 h in the WT. The photosynthetic rate noticeably decreased only in LA4012 and LA3538 after 24 h of irradiation. After 72 h, the photosynthetic rate decreased in all mutants, with the exception of hp-1.2 LA0279, but the decrease was most noticeable in LA4012, yet significant changes in the respiration rate were absent. The LA0279 mutant was more capable of accumulating anthocyanin in the cells of the subepidermal parenchyma and chlorenchyma, as well as in the cells at the base of large multicellular glandular trichomes and in the mesophyll. Another important difference was the accumulation of increased amounts of antheraxanthin and phenolic compounds in the leaves of LA0279 after 72 h of HIL irradiation. Unlike LA4012, LA3006, LA0279, and LA3538 sowed a significant increase in the expression levels of CHS, HY5, and FLS genes after 24 h, which may be one of the reasons for the higher adaptive potential of those three mutants. In addition to that in LA3538, strong light-induced stress led to an increased level of flavonol synthase (FLS) expression in the LA3006, LA0279, and LA4012 mutants. We hypothesize that the photosynthetic apparatus (PA) of the LA0279 mutant, which is deficient in the DET1 and DDB1 genes, is most adapted to prolonged HIL. Most likely, the resistance of PA mutants to HIL is due to a variety of factors, which, in addition to the redistribution of carotenoids, may include morphological features associated with the accumulation of anthocyanin in the epidermis, subepidermal layer, mesophyll and trichomes of leaves and with an increase in leaf thickness.

The skin represents the largest area for direct contact between microbes and host immunocytes and is a site for constant communication between the host and this diverse and essential microbial community. Coagulase-negative staphylococci are an abundant bacterial genus on the human skin and are regulated through various mechanisms that include the epidermal barrier environment and innate and adaptive immune systems within the epidermis and dermis. In turn, some species and strains of these bacteria produce beneficial products that augment host immunity by exerting specifically targeted antimicrobial, anti-inflammatory, or anti-neoplastic activity while also promoting broad innate and adaptive immune responses. The use of selected skin commensals as a therapeutic has shown promise in recent human clinical trials. This emerging concept of bacteriotherapy is defining mechanisms of action and validating the dependence on the microbiome for maintenance of immune homeostasis.

Objective To evaluate the quality of Arnebiae Radix (AR) and Dictamni Cortex (DC) and study the efficacy of herbal extracts of these two herbs on the treatment of allergic contact dermatitis (ACD). Methods Qualitative and quantitative analysis of effective components was performed using High Performance Thin Layer Chromatography (HPTLC), High Performance Liquid Chromatography (HPLC), and HPLC-Quadrupole Time of Flight-Mass Spectrometry (HPLC-QTOF-MS). In vitro allergic ACD 3D model was established by incubating 3D reconstructed human epidermis (RHE) with skin sensitizer, potassium dichromate. A total of 65 gene expression that were associated with ACD, which included 24 antioxidant responsive element (ARE) and 41 SENS-IS genes were quantified by qRT-PCR. More than or equal to 10 ARE genes and 18 SENN-IS genes were induced by 1.3-fold, demonstrating the successful establishment of in vitro ACD model. Oil extracts of AR and DC were applied on the in vitro ACD model to study the efficacy. Results Batch 3 of AR and batch 2 of DC showed presence of all active ingredients with the highest concentrations. Active ingredients of the herbs were extracted using a special oil and formulated into herbal oil extracts. The herbal oil extracts were able to down regulate the induced genes in the in-vitro ACD skin model, bringing the tissue back to homeostatic status. Conclusion The oil extracts showed the potent efficacy of using AR and DC in ACD treatment. The combination study will be done to optimize the formulation ratio which will be developed into a topical cream.

In Arabidopsis thaliana, the epidermis is the outermost cell layer composed of many specialized types of epidermal cells, such as pavement cells, trichomes, and guard cells. The homeodomain-leucine zipper (HD-ZIP) class Ⅳ transcription factors (TFs), which are unique to the plant kingdom, have been recognized as key regulators of epidermis development. Unlike animal HD proteins, which can bind to DNA as monomers, plant HD-ZIP class Ⅳ TFs bind to DNA as dimers, although little is known about the regulation of their dimerization process. Here, we show that the homodimerization of ARABIDOPSIS THALIANA MERISTEM LAYER 1 (ATML1) - HD-ZIP class Ⅳ TF that is required for protoderm development - is regulated by the lipid-binding steroidogenic acute regulatory protein-related lipid transfer (START) domain. We found that ATML1 forms homodimer through interaction via its ZIP motif in yeast and plant cells, although the interaction is abolished by generating a mutation into the lipid-binding START domain to disrupt the lipid-binding ability. These results suggest that lipidic ligands function as key regulators of protoderm development via modulating the dimerization of ATML1.

Atopic dermatitis (AD) is a chronic inflammatory skin disease that affects up to one in five children and millions of adults in developed countries. Clinically, AD skin lesions manifest as subacute and/or chronic lichenified eczematous plaques, which are often intensely pruritic and prone to secondary bacterial and viral infections. Despite the emergence of novel therapeutic agents, treatment options and outcomes for AD remain suboptimal. An improved understanding of AD pathogenesis may help improve patient outcomes. Dysregulated Th2-polarized skin inflammation and impaired skin barrier function interact to drive AD pathogenesis; however, much remains to be understood about the molecular mechanisms underlying this interplay. The current study used published clinical trial datasets to define a skin-related AD gene signature. This meta-analysis revealed significant reductions in IL1F7 transcripts (encodes IL-37) in AD patient samples. Reduced IL1F7 correlated with lower transcripts for key skin barrier function genes in the epidermal differentiation complex. Immunohistochemical analysis of normal (healthy) human skin specimens and an in vitro three-dimensional human skin model localized IL-37 protein to the epidermis. In comparison with normal human skin, IL-37 levels were decreased in AD patient skin. Addition of Th2 cytokines to the aforementioned in vitro three-dimensional skin model recapitulates key aspects of AD skin and was sufficient to reduce epidermal IL-37 levels. Image analysis also indicated close relationship between epidermal IL-37 and skin epidermal differentiation complex proteins. These findings suggest IL-37 is intimately linked to normal keratinocyte differentiation and barrier function and implicates IL-37 as a potential biomarker and therapeutic target for AD.

Background Keratinocytes and fibroblasts represent the major cell types in the epidermis and dermis of the skin and play a significant role in maintenance of skin homeostasis. However, the biological characteristics of keratinocytes and fibroblasts remain to be elucidated. The purpose of this study was to compare the gene expression pattern between keratinocytes and fibroblasts and to explore novel biomarker genes so as to provide potential therapeutic targets for skin-related diseases such as burns, wounds, and aging. Methods Skin keratinocytes and fibroblasts were isolated from newborn mice. To fully understand the heterogeneity of gene expression between keratinocytes and fibroblasts, differentially expressed genes (DEGs) between the two cell types were detected by RNA-seq technology. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the known genes of keratinocytes and fibroblasts and verify the RNA-seq results. Results Transcriptomic data showed a total of 4309 DEGs (fold-change > 1.5 and q-value

Upon skin injury, re-epithelialization must be triggered promptly to restore the integrity and barrier function of the epidermis. However, this process is often delayed or interrupted in chronic wounds like diabetic foot ulcers. Considering that BG particles can activate multiple genes in various cells, herein, we hypothesized that bioactive glass (BG) might be able to modulate the barrier functional behaviors of keratinocytes. By measuring the transepithelial electrical resistance (TEER) and the paracellular tracer flux, we found the 58S-BG extracts substantially enhanced the barrier function of keratinocyte monolayers. The BG extracts might exert such effects by promoting the keratinocyte differentiation and the formation of tight junctions, as evidenced by the increased expression of critical differentiation markers (K10 and involucrin) and TJ protein claudin-1, as well as the altered subcellular location of four major TJ proteins (claudin-1, occludin, JAM-A, and ZO-1). Besides, the cell scratch assay showed that BG extracts induced the collective migration of keratinocytes, though they did not accelerate the migration rate compared to the control. The in vivo study using a diabetic rat wound model demonstrated that the BG extracts accelerated the process of re-epithelialization, stimulated keratinocyte differentiation, and promoted the formation of tight junctions in the newly regenerated epidermis. Our findings revealed the crucial effects of BGs on keratinocytes and highlighted its potential application for chronic wound healing by restoring the barrier function of the wounded skin effectively., Graphical abstract Image 1, Highlights • Re-epithelialization is often delayed or interrupted in chronic wounds • The 58S-BG extracts induced the collective migration of keratinocytes. • The 58S-BG extracts regulated the barrier functional behaviors of keratinocytes by promoting the keratinocyte differentiation and the formation of tight junctions.

Currently, human-skin derived cell culture is a basic technique essential for dermatological research, cellular engineering research, drug development, and cosmetic development. But the number of donors is limited, and primary cell function reduces through cell passage. In particular, since adult stem cells are present in a small amount in living tissues, it has been difficult to obtain a large amount of stem cells and to stably culture them. In this study, skin derived cells were isolated from the epidermis, dermis, and adipose tissue collected from single donor, and immortalization was induced through gene transfer. Subsequently, cell lines that could be used as stem cell models were selected using the differentiation potential and the expression of stem cell markers as indices, and it was confirmed that these could be stably cultured. The immortalized cell lines established in this study have the potential to be applied not only to basic dermatological research but also to a wide range of fields such as drug screening and cell engineering.

Polyploid plants often show improved resistance against many diseases, but whether they show increased resistance to grey mould, a devastating disease caused by Botrytis spp. fungi, is seldom reported. Stomata and reactive oxygen species (ROS) play dual roles in defence against grey mould, and it is unclear how their roles change after polyploidization. We addressed these questions in diploid and colchicine-induced Lilium rosthornii after B. elliptica infection. Tetraploids were less susceptible to grey mould, with lower morality rates in naturally infected plants. Before the stomata closed in artificially infected leaves, tetraploids, with larger stomatal apertures, were more easily invaded by the pathogen than diploids. However, the lesion area increased more slowly in tetraploids than in diploids, which may be explained by three causes based on histological and physiological characteristics. First, the pathogen required more time to penetrate the epidermis and closed stomata in tetraploids than in diploids. Second, the pathogen penetrated the reopened stomata more easily than the epidermis, and stomatal density was lower in tetraploids than in diploids. Third, tetraploids showed faster ROS accumulation, a more effective ROS-scavenging system and less malondialdehyde (MDA) accumulation than diploids. Stomatal starch and abnormal guard cell nuclei were present in the infected leaves. This phenomenon may be caused by oxalic acid, a pathogenic factor for many pathogens that promotes stomatal starch degradation and stomatal reopening in Sclerotinia spp., a pathogen closely related to Botrytis spp. This suggestion was primarily confirmed by immersing healthy leaves in oxalic acid solution.

Syzygium cumini (L.) Skeels (Family: Myricaceae) commonly known as Jamun is a widely used medicinal plant in Ayurveda. In spite of its numerous medicinal attributes, no published work is available on pharmacognostic characterization, physicochemical analysis of its leaves. The measures taken for pharmacognostic characterization were macroscopy, microscopy, powder microscopy, physicochemical evaluation, fluorescence analysis, and preliminary phytochemical screening. The microscopic evaluation showed useful characters for the recognition of leaf of Syzygium cumini (L.) Skeels . Transverse section, showed presence of single layer of wavy epidermal cells with striated cuticle single layer Palisade beneath upper epidermis in lamina region (contains compact elongated cells), Spheraphide in lamina region, collenchyma below the upper epidermis and above the lower epidermis in midrib region, Xylem and phloem in center, Sclerenchyma in between vascular bundle and collenchyma in midrib region. Wavy epidermal cell in the upper epidermis, anisocytic stomata in the lower epidermis, Bordered pitted reticulate vessels, Calcium oxalate crystal sheath in the lamina, fragment of fibers and Spiral vessels were observed in powder microscopy. Physicochemical studies showed total ash (3.1%), acid insoluble ash (0.7%), alcohol soluble extractive values (10.96 %), and water-soluble extractive values (12.32 %). Phytochemical analysis revealed the presence of sugar, lipid, Glycoside, saponins, phenols, flavonoids, tannins, tri-terpenoids and steroids. The study forms the first report on pharmacognostic characters and a physicochemical parameter which could be useful for identification and authentication of the plant.

Background Chronic exposure to ultraviolet (UV) radiation promotes skin photoaging, which is clinically characterized by dryness, laxity, and wrinkling. Sea cucumber (Stichopus japonicus) (SC) is a marine organism with culinary and medicinal applications, especially in Asian countries. Additionally, SC is a potential nutraceutical as it exhibits bioactivities, such as antioxidant, antitumor, and anticancer activities. This study examined the effects of SC and its hydrolysate (SCH) on UVA-induced skin barrier function and wrinkle formation using hairless mice. Results UVA significantly induced transepidermal water loss and wrinkle formation, which were significantly mitigated upon oral administration of SC and SCH. Additionally, SC mitigated the UVA-induced downregulation of epidermal natural moisturizing factors and upregulation of Aqp3, Mmp13, Tnfa, and Il6 mRNA levels in the mouse skin. Conclusion Taken together, these results suggest that dietary SC and SCH exert anti-photoaging effects by modulating filaggrin synthesis and desquamation in the epidermis and regulating the NF-κB pathway in the skin. Our research indicates that SC and SCH have potential applications in nutricosmetics for photoaging. This article is protected by copyright. All rights reserved.

Vulval extramammary Paget disease (vEMPD) is an uncommon epithelial malignancy that may arise within the vulva (primary vEMPD) or represent vulval skin involvement by a noncutaneous carcinoma (secondary vEMPD). Primary vEMPD is most often an in situ carcinoma arising within the epidermis but may be associated with dermal invasion (invasive vEMPD) or represent intraepidermal spread of an adenocarcinoma originating in vulval skin adnexa or anogenital mammary-like glands. The latter, termed mammary gland-like adenocarcinoma (MGLA), exhibits morphologic, immunohistochemical, and molecular features of various breast carcinomas but, as far as we are aware, the metaplastic variant of MGLA has not been reported on the vulva. We report 2 cases of metaplastic MGLA of the vulva with associated Paget disease and postulate that some cases of vulval MGLA may arise from Paget disease rather than originating in mammary-like glands. We also report a unique case of secondary vEMPD resulting from spread of urothelial carcinoma in situ that subsequently progressed to invasive urothelial carcinoma within the vulva.

Para verificar a influência da altitude na estrutura anatômica de folhas e do xilema secundário de galhos de indivíduos da espécie Clethra scabra Pers. (Clethraceae) ocorrentes na Serra da Mantiqueira, estado de Minas Gerais, realizaram-se coletas de amostras nas cotas altitudinais de 1500, 1700, 1900 e 2100 m (metros). As folhas e galhos coletados foram fixados e preparados seguindo procedimentos usuais de microtécnica vegetal. Procederam-se análises anatômicas quantitativas utilizando software de análise de imagem. As folhas de C. scabra são hipoestomáticas com epiderme trisseriada e tricomas estrelados, mesofilo dorsiventral, parênquima paliçádico com uma a duas camadas celulares e o parênquima esponjoso com quatro. A nervura central com xilema desenvolvido e o feixe vascular envolvido pela bainha esclerenquimática. Análises de variância (ANOVA) e teste de Scott-Knott demonstraram que as espessuras do limbo foliar, mesofilo e parênquima paliçádico não apresentaram diferenças estatísticas significativas entre as altitudes avaliadas. O xilema secundário apresentou diminuição no comprimento dos elementos de vaso com o aumento da altitude na cota de 2100 m e o diâmetro tangencial dos elementos de vaso aumentaram nas duas menores altitudes e depois reduziram. Índices de vulnerabilidade de Carlquist e de mesomorfia diminuíram ao longo do gradiente altitudinal. Comprimento e diâmetro das fibras diminuíram com elevação da altitude. Assim, a altitude influencia nas modificações anatômicas apresentadas por folhas e galhos de C. scabra, que podem ser relacionadas a sua plasticidade fenotípica em resposta às diferentes condições ambientais.

The epidermis of mouse skin is usually populated by dendritic epidermal T cells (γδDETC) expressing an invariant Vγ5Vδ1+ TCR. In Tcrd-/- mice, skin-resident γδDETC are replaced by αβDETC carrying polyclonal αβ TCRs. Although they exhibit a dendritic morphology, αβDETC were reported to be less functional than genuine γδDETC, likely because their TCR is unable to interact with the original TCR ligands of γδDETC. However, the TCR repertoire of those replacement DETC in Tcrd-/- mice might provide clues for understanding the development and selection of canonical γδDETC. Here, we compare the phenotype and TCR repertoires of wild-type and Tcrd-/- mouse skin T cells. Our data reveal that αβDETC are CD4/CD8 double negative and express an MHC-independent TCR repertoire. Furthermore, we identify a second MHC-independent population of CD103hi CD4/ CD8 double-negative αβ T cells in the dermis of Tcrd-/- mice.

Salt stress reduces plant growth by negatively interfering with the division rate and cellular expansion, limiting the growth and development of the roots, stems, and leaves. 24-Epibrassinolide (EBR) is a molecule extracted from plant tissues and is a plant growth regulator with a high capacity to modulate tolerance to abiotic stresses. The objective of this study was to verify the possible improvements promoted by pretreatment with EBR in salt-stressed tomato plants, evaluating the variables related to root anatomy, photosynthetic pigments, antioxidant system, and biomass accumulation. The experiment comprised four treatments: two salt conditions (0 and 150 mM NaCl, described as Na+ (−) and Na+ ( +), respectively) and two concentrations of 24-epibrassinolide (0 and 100 nM EBR, described as EBR (−) and EBR ( +), respectively). EBR modulated the protection and vascularization of root structures, as demonstrated by the increases in epidermis thickness (12%) and metaxilem diameter (119%), respectively. This steroid relieved oxidative damage, which was clearly linked to elevated activities of superoxide ascorbate peroxidase (24%) and guaiacol peroxidase (31%). EBR also benefited photosynthetic pigments, reducing the degradation of chlorophylls. In addition, pretreatment with EBR favoured a higher biomass, which was due to positive effects on leaf and root tissues, including better performance of photosynthetic machinery.

Background Maximum cyclobutane pyrimidine dimer (CPD) formation in the skin induced by ultraviolet B (UVB) irradiation is thought to occur within a few minutes and is immediately decreased by the DNA repair system. Objective We evaluated the time course and differential effects of narrowband (NB-UVB) and broadband (BB-UVB) UVB on CPD formation. Methods We investigated CPD formation at various time-points in vivo, from 3 min to 72 h, after UVB irradiation using 2 mouse strains, C57BL/6 J and BALB/c. The backs of the mice were shaved and irradiated with NB-UVB or BB-UVB. Skin specimens were obtained and stained with anti-CPD antibody. Positive signals in the epidermis were measured using ImageJ. DNA was extracted from the isolated epidermis and subjected to quantitative CPD analysis by enzyme-linked immunosorbent assay (ELISA). Results CPDs induced by UVB irradiation (1 minimum erythemal dose) in epidermal skin were detected in the nucleus. Although the CPD levels increased immediately after irradiation (3 min), the highest level was detected at 1 h and the increase lasted 24–48 h after irradiation. BB-UVB tended to induce greater CPD levels than NB-UVB in both mouse strains. The ELISA showed similar results. Conclusions CPDs were induced immediately after UV irradiation, with the maximum level observed 1 h after irradiation. BB-UVB irradiation tended to induce greater levels of CPD formation. In addition to the direct effects of UVB, the presence of CPDs in hair follicles, which were not irradiated by UVB, suggests that reactive oxygen species are also involved in CPD formation in the skin.

Innate-like T cells, including invariant NKT cells, mucosal-associated invariant T (MAIT) cells, and γ δ T (γδT) cells, are groups of unconventional T lymphocytes. They play important roles in the immune system. Because of the lack of Cre recombinase lines that are specific for innate-like T cells, pan–T cell Cre lines are often used to study innate-like T cells. In this study, we found that distal Lck promoter–driven Cre (dLckCre) in which the distal Lck gene promoter drives Cre expression in the late stage of thymocyte development has limited function in the innate-like T cells using ROSA26floxed-Stop-tdTomato reporter. Innate-like T cells differentiate into mature functional subsets comparable to the CD4+ Th subsets under homeostatic conditions. We further showed that dLckCre-expressing γδT cells are strongly biased toward γδT1 phenotype. Interestingly, the γδT cells residing in the epidermis and comprising the vast majority of dendritic epidermal T cells nearly all express dLckCre, indicating dLckCre is a useful tool for studying dendritic epidermal T cells. Taken together, these data suggest that Lck distal promoter has different activity in the conventional and unconventional T cells. The use of dLCKcre transgenic mice in the innate-like T cells needs to be guided by a reporter for the dLckCre function.

Objective: We compared the capability of human umbilical vein endothelial cells (HUVECs), induced pluripotent stem cell (iPSC)-derived endothelial cells (iECs), and human dermal blood endothelial cells (HDBECs) to effectively vascularize engineered human skin constructs (HSCs) in vitro and on immunodeficient mice. Approach: We quantified the angiogenesis within HSCs both in vitro and in vivo through computational analyses of immunofluorescent (IF) staining. We assayed with real-time quantitative PCR (RT-qPCR) the expression of key endothelial, dermal, and epidermal genes in 2D culture and HSCs. Epidermal integrity and proliferation were also evaluated through haematoxylin and eosin staining, and IF staining. Results: IF confirmed iEC commitment to endothelial phenotype. RT-qPCR showed HUVECs and iECs immaturity compared with HDBECs. In vitro, the vascular network extension was comparable for HDBECs and HUVECs despite differences in vascular diameter, whereas iECs formed unorganized rudimentary tubular structures. In vivo, all ECs produced discrete vascular networks of varying dimensions. HUVECs and HDBECs maintained a higher proliferation of basal keratinocytes. HDBECs had the best impact on extracellular matrix expression, and epidermal proliferation and differentiation. Innovation: To our knowledge, this study represents the first direct and quantitative comparison of HDBECs, HUVECs, and iECs angiogenic performance in HSCs. Conclusions: Our data indicate that HUVECs and iECs can be an alternative cell source to HDBEC to promote the short-term viability of prevascularized engineered grafts. Nevertheless, HDBECs maintain their capillary identity and outperform other EC types in promoting the maturation of the dermis and epidermis. These intrinsic characteristics of HDBECs may influence the long-term function of skin grafts.