Tissue Migration Research Articles

Investigation of Post‐Translational Modification on NHE1 Transport Function

The sodium hydrogen exchanger isoform 1 (NHE‐1) is a sodium hydrogen antiporter that regulates intracellular pH (pHi), cell volume, dynamic actin remodeling processes, and coordination of cell migration in mammalian tissues. NHE‐1 is post‐translationally modified by phosphorylation and lipid modification (palmitoylation). The intracellular C‐terminal domain of NHE‐1 is reversibly phosphorylated at multiple sites byseveral kinases including: extracellular signal‐regulated kinase (ERK), protein kinase B (AKT), B‐Raf, p90 Ribosomal S6 Kinase (p90rsk), RhoA Kinase (ROCK), and other protein kinases. We recently identified that NHE‐1 is reversibly palmitoylated (S‐acetylation), in which a 16‐carbon palmitic acid is covalently attached to a cysteine residue on the C‐terminal domain via a thioester linkage. Such modifications are reported to regulate protein trafficking, membrane micro localization, and protein‐protein interactions. Steady‐state pHi assays incubated with pH‐sensitive dye were conducted to measure the change in pHi of Chinese Hamster Lung Fibroblast Cells due to NHE‐1. Investigating agonists and stimulating factors that increase NHE‐1 palmitoylation allows us to determine the impact of palmitoylation on NHE‐1 transport. Palmitoyl‐transferases (PATs) catalyze the lipidation of proteins and are inhibited by 2‐bromo‐palmitate (2BP). The presence of 2BP causes a loss of palmitoylation to occur, effectively inhibiting LPA‐induced NHE‐1 activity by 0.1 pH units. Fetal Bovine Serum (FBS), Lysophosphatidic acid (LPA), Phorbol 12‐myristate 13‐acetate (PMA), and Insulin effectively stimulate NHE‐1 by increasing pHi by 0.1 pH units. In the presence of kinase inhibitors, NHE‐1 activity significantly decreases compared to agonist or serum treatment alone. The impact of the combination of kinase inhibitors and palmitoylation inhibitors on NHE‐1 mediated pHi change was determined. The data supports the hypothesis that palmitoylation and phosphorylation coordinate to subtly modify NHE1 activity in a novel rheostat of the cell.

Early Dendritic Morphogenesis of Adult-Born Dentate Granule Cells Is Regulated by FHL2.

Dentate granule cells (DGCs), the progeny of neural stem cells (NSCs) in the sub-granular zone of the dentate gyrus (DG), must develop and functionally integrate with the mature cohort of neurons in order to maintain critical hippocampal functions throughout adulthood. Dysregulation in the continuum of DGC development can result in aberrant morphology and disrupted functional maturation, impairing neuroplasticity of the network. Yet, the molecular underpinnings of the signaling involved in adult-born DGC maturation including dendritic growth, which correlates with functional integration, remains incompletely understood. Given the high metabolic activity in the dentate gyrus (DG) required to achieve continuous neurogenesis, we investigated the potential regulatory role of a cellular metabolism-linked gene recently implicated in NSC cycling and neuroblast migration, called Four and a half LIM domain 2 (FHL2). The FHL2 protein modulates numerous pathways related to proliferation, migration, survival and cytoskeletal rearrangement in peripheral tissues, interacting with the machinery of the sphingosine-1-phosphate pathway, also known to be highly active especially in the hippocampus. Yet, the potential relevance of FHL2 to adult-born DGC development remains unknown. To elucidate the role of FHL2 in DGC development in the adult brain, we first confirmed the endogenous expression of FHL2 in NSCs and new granule cells within the DG, then engineered viral vectors for genetic manipulation experiments, investigating morphological changes in early stages of DGC development. Overexpression of FHL2 during early DGC development resulted in marked sprouting and branching of dendrites, while silencing of FHL2 increased dendritic length. Together, these findings suggest a novel role of FHL2 in adult-born DGC morphological maturation, which may open up a new line of investigation regarding the relevance of this gene in physiology and pathologies of the hippocampus such as mesial temporal lobe epilepsy (MTLE).

Anti-inflammatory and antioxidant activity improvement of lycopene from guava on nanoemulsifying system

The nanoemulsion system can improve stability to antioxidant molecules when compared to the isolated molecule. Therefore, this study aimed to establish conditions on thermal stability, chemical characterization, antioxidant and anti-inflammatory activity of lycopene nanoemulsion. Different chemical assays were used to characterize the lycopene nanoemulsion (LN) and the purified lycopene from guava (LPG) as HPLC, UV-Vis, DLS, and ABTS to analyze the antioxidant activity of lycopene nanoemulsion. To test its thermal stability as an antioxidant in nanoemulsion it was used the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) techniques. The carrageenan-induced paw edema model in Swiss mice and histopathological analysis were used to evaluate anti-inflammatory activity. Emulsion with a mean droplet size of z-Average diameter 167.1 d.nm was produced. UV-vis detected λmax = 447.1, 473.7, and 505.3 nm to lycopene from guava and HPLC-DAD analysis by C30 at 472 nm identified the presence of lycopene in the nanoemulsion. This lycopene nanoemulsion presented thermal stability at room temperature, as well as on concentration when stored for one year. Also significantly inhibited (p < 0.05) the edema formation, with maximum inhibitory effect at 50 mg/kg by oral and intraperitoneal route and reduced neutrophil migration in paw tissue. The lycopene nanoemulsion can be a potential candidate for investigation and therapy of various diseases, including inflammation because this lycopene nanoemulsion presented therapeutics application for inflammatory diseases, and has also potential to be used as ointment and/or cream additive for topic applications.

Genetic engineering of Hoxb8-immortalized hematopoietic progenitors - a potent tool to study macrophage tissue migration.

Tumor-associated macrophages (TAMs) are detrimental in most cancers. Controlling their recruitment is thus potentially therapeutic. We previously found that TAMs perform protease-dependent mesenchymal migration in cancer, while macrophages perform amoeboid migration in other tissues. Inhibition of mesenchymal migration correlates with decreased TAM infiltration and tumor growth, providing rationale for a new cancer immunotherapy specifically targeting TAM motility. To identify new effectors of mesenchymal migration, we produced ER-Hoxb8-immortalized hematopoietic progenitors (cells with estrogen receptor-regulated Hoxb8 expression), which show unlimited proliferative ability in the presence of estrogen. The functionality of macrophages differentiated from ER-Hoxb8 progenitors was compared to bone marrow-derived macrophages (BMDMs). They polarized into M1- and M2-orientated macrophages, generated reactive oxygen species (ROS), ingested particles, formed podosomes, degraded the extracellular matrix, adopted amoeboid and mesenchymal migration in 3D, and infiltrated tumor explants ex vivo using mesenchymal migration. We also used the CRISPR/Cas9 system to disrupt gene expression of a known effector of mesenchymal migration, WASP (also known as WAS), to provide a proof of concept. We observed impaired podosome formation and mesenchymal migration capacity, thus recapitulating the phenotype of BMDM isolated from Wasp-knockout mice. Thus, we validate the use of ER-Hoxb8-immortalized macrophages as a potent tool to investigate macrophage functionalities.

Characterization of Glyceollins as Novel Aryl Hydrocarbon Receptor Ligands and Their Role in Cell Migration.

Recent studies strongly support the use of the aryl hydrocarbon receptor (AhR) as a therapeutic target in breast cancer. Glyceollins, a group of soybean phytoalexins, are known to exert therapeutic effects in chronic human diseases and also in cancer. To investigate the interaction between glyceollin I (GI), glyceollin II (GII) and AhR, a computational docking analysis, luciferase assays, immunofluorescence and transcriptome analyses were performed with different cancer cell lines. The docking experiments predicted that GI and GII can enter into the AhR binding pocket, but their interactions with the amino acids of the binding site differ, in part, from those interacting with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Both GI and GII were able to weakly and partially activate AhR, with GII being more potent. The results from the transcriptome assays showed that approximately 10% of the genes regulated by TCDD were also modified by both GI and GII, which could have either antagonistic or synergistic effects upon TCDD activation. In addition, we report here, on the basis of phenotype, that GI and GII inhibit the migration of triple-negative (ER-, PgR-, HER2NEU-) MDA-MB-231 breast cancer cells, and that they inhibit the expression of genes which code for important regulators of cell migration and invasion in cancer tissues. In conclusion, GI and GII are AhR ligands that should be further investigated to determine their usefulness in cancer treatments.

Milk Fat Globule-EGF Factor 8 Contributes to Progression of Hepatocellular Carcinoma.

Milk fat globule-EGF factor 8 (MFG-E8) is an anti-inflammatory glycoprotein that mediates a wide spectrum of pathophysiological processes. MFG-E8 has been studied as a key regulator of cancer cell invasion, migration, and proliferation in different tissues and organs. However, potential roles of MFG-E8 in the growth and progression of liver cancer have not been investigated to date. Here, we analyzed 33 human hepatocellular carcinoma (HCC) samples and found that levels of MFG-E8 expression were significantly higher in HCC cells than in normal liver tissues. In addition, our in vitro gain-of-function study in three different HCC cell lines revealed that overexpression of MFG-E8 promoted the proliferation and migration of HCC cells, as determined by RT-qPCR, MTT assays, and wound healing analyses. Conversely, an MFG-E8 loss-of function study showed that proliferation capacity was significantly reduced by MFG-E8 knockdown in HCC cells. Additionally, MFG-E8 activity-neutralizing antibodies profoundly inhibited both migration and proliferation of HCC cells, attenuating their tumorigenic properties. These reductions in migration and proliferation were rescued by treatment of HCC cells with recombinant MFG-E8 protein. Furthermore, an in vivo HCC xenograft study showed that the number of proliferating HCC cells and tumor volume/weight were all significantly increased by MFG-E8 overexpression, compared to control mice. These results clearly show that MFG-E8 plays an important role in HCC progression and may provide a basis for future mechanistic studies and new strategies for the treatment of liver cancer.

Dynamic Buffering of Extracellular Chemokine by a Dedicated Scavenger Pathway Enables Robust Adaptation during Directed Tissue Migration.

How tissues migrate robustly through changing guidance landscapes is poorly understood. Here, quantitative imaging is combined with inducible perturbation experiments to investigate the mechanisms that ensure robust tissue migration invivo. We show that tissues exposed to acute "chemokine floods" halt transiently before they perfectly adapt, i.e., return to the baseline migration behavior in the continued presence of elevated chemokine levels. A chemokine-triggered phosphorylation of the atypical chemokine receptor Cxcr7b reroutes it from constitutive ubiquitination-regulated degradation to plasma membrane recycling, thus coupling scavenging capacity to extracellular chemokine levels. Finally, tissues expressing phosphorylation-deficient Cxcr7b migrate normally in the presence of physiological chemokine levels but show delayed recovery when challenged with elevated chemokine concentrations. This work establishes that adaptation to chemokine fluctuations can be "outsourced" from canonical GPCR signaling to an autonomously acting scavenger receptor that both senses and dynamically buffers chemokine levels to increase the robustness of tissue migration.

Sphere-forming corneal cells repopulate dystrophic keratoconic stroma: Implications for potential therapy.

Keratoconus is a degenerative corneal disease characterised by aberrant cell behaviour and loss of matrix that can result in vision loss. Cells extracted from peripheral corneas can form stem cell-enriched spheres, which have shown the potential to repopulate the normal peripheral corneal stroma in vitro upon sphere implantation but have not been previously studied in keratoconic tissue. To investigate the therapeutic potential of stem cell-enriched spheres formed from extracted peripheral human corneal cells when introduced to keratoconic tissue. Stem cell-enriched spheres were formed from extracts of normal cadaveric human peripheral corneal cells. These spheres were implanted into incisions created in full thickness and onto the surface of 10 µm thin sections of keratoconic and normal stromal tissues in vitro. Tissue sections were used to maximise use of limited keratoconic tissue available for research. Living cells were stained with Calcein-AM and visualised with stereo and fluorescence microscopy to assess survival and behaviours between the time of implantation day 0 and 14 d (D14) from implantation. Sphere cells in implanted tissues were characterised for stem cell and differentiation markers using immunohistochemistry and droplet digital PCR to assess the potential implications of these characteristics in the use of spheres in keratoconus treatment. Spheres were successfully implanted into full-thickness central corneal tissue and onto the surface of 10 µm thin en face tissue sections. No observable differences were seen in sphere migration, proliferation or differentiation in keratoconic tissue compared to normal between day 0 and D14. Spheres stained positively with Calcein-AM up to D14. Cell migration increased from day 0 to D14, occurring radially in three dimensions from the sphere and in alignment with tissue edges. Cell proliferation marker, EdU, was detected at day 10. Implanted spheres stained positively for putative stem cell markers ∆Np63α and ABCB5, while ABCG2, ABCB5, ∆Np63 and p63α were detectable by droplet digital PCR up to D14. Double immunolabelling revealed absence of ABCB5 staining in migrated cells but positive staining of alpha smooth muscle actin (myofibroblast marker) in some migrated cells. Droplet digital PCR showed similar expression patterns of differentiation markers but a reduction in stem cell markers between normal and keratoconic tissue with an increase in stromal cell markers and a reduction in epithelial cell markers, indicating an appropriate response to repopulating diseased tissue. Cells from implanted stem cell-enriched spheres can repopulate a keratoconic corneal stromal surface in a directed manner and exhibit migratory stromal cell phenotypes.

Monte Carlo Modeling of Near-infrared Fluorescence Photon Migration in Breast Tissue for Tumor Prediction

Monte Carlo Modeling of Near-infrared Fluorescence Photon Migration in Breast Tissue for Tumor Prediction

Hyperdry human amniotic membrane application as a wound dressing for a full-thickness skin excision after a third-degree burn injury.

Background Severe burn injuries create large skin defects that render the host susceptible to bacterial infections. Burn wound infection often causes systemic sepsis and severe septicemia, resulting in an increase in the mortality of patients with severe burn injuries. Therefore, appropriate wound care is important to prevent infection and improve patient outcomes. However, it is difficult to heal a third-degree burn injury. The aim of this study was to investigate whether hyperdry human amniotic membrane (HD-AM) could promote early granulation tissue formation after full-thickness skin excision in third-degree burn injury sites in mice.Methods After the development of HD-AM and creation of a third-degree burn injury model, the HD-AM was either placed or not placed on the wound area in the HD-AM group or HD-AM group, respectively. The groups were prepared for evaluation on postoperative days 1, 4 and 7. Azan staining was used for granulation tissue evaluation, and estimation of CD163, transforming growth factor beta-1 (TGF-β1), vascular endothelial growth factor (VEGF), CD31, alpha-smooth muscle actin (α-SMA) and Iba1 expression was performed by immunohistochemical staining. Quantitative reverse-transcription polymerase chain reaction (PCR) was used to investigate gene expression of growth factors, cell migration chemokines and angiogenic and inflammatory markers.ResultsThe HD-AM group showed significant early and qualitatively good growth of granulation tissue on the full-thickness skin excision site. HD-AM promoted early-phase inflammatory cell infiltration, fibroblast migration and angiogenesis in the granulation tissue. Additionally, the early infiltration of cells of the immune system was observed.Conclusions HD-AM may be useful as a new wound dressing material for full-thickness skin excision sites after third-degree burn injuries, and may be a new therapeutic technique for improving the survival rate of patients with severe burn injuries.

Tractionless Self-Propulsion of Active Drops

We report on a new mode of self-propulsion exhibited by compact drops of active liquids on a substrate which, remarkably, is tractionless, i.e., which imparts no mechanical stress locally on the surface. We show, both analytically and by numerical simulation, that the equations of motion for an active nematic drop possess a simple self-propelling solution, with no traction on the solid surface and in which the direction of motion is controlled by the winding of the nematic director field across the drop height. The physics underlying this mode of motion has the same origins as that giving rise to the zero viscosity observed in bacterial suspensions. This topologically protected tractionless self-propusion provides a robust physical mechanism for efficient cell migration in crowded environments like tissues.

Development of an In-body-tissue-engineered Collagenous Biosheet for Regenerative Vascular Reconstruction in a Beagle Model

Introduction - Autologous pericardial tissues are commonly used for the arterial reconstruction. To overcome their limited amount, tissue-engineered material that has potential to engraft to native tissues and then grow is desired especially in pediatric surgery. In body tissue architecture (iBTA) can prepare autologous collagenous sheet-like tissues, called Biosheets, by using a patient's body as a bioreactor. Its process is simple and safe, just subcutaneously embedding a mold for about 2 months. Since Biosheet has sufficient mechanical properties they could be applied as reconstruction materials for abdominal wall or diaphragm. This study investigated the possibility of Biosheet as a regenerative vascular reconstruction material in a beagle model. Methods - Cylindrical molds (diameter 18 mm, length 7 cm) were embedded in beagles or cows (Hokkaido University) subcutaneously for 2 months. Tubular tissues were formed inside of all molds, and were harvested upon removing the mold. Canine or bovine Biosheets (size 5 x 7 cm, thickness ca. 0.6 mm) were obtained by cutting the tubular tissues at the longitudinal direction as collagenous sheet-like tissues. Arterial defects at pulmonary arteries (ca. 10 × 20 mm, 1 in photos) or descending aorta (ca. 10 x 10 mm, 2 in photos) were made and patch repair using Biosheets was performed in beagles under partial clamping. Results - In ultrasonic echo observation, little change in blood flow change occurred after the patching. Angiogram showed no elongation or stenosis at the patched region (3 in photos). Tissue migration from native artery occurred primarily on the luminal surface of the Biosheets with their degradation (4 in photos). Even in xeno implantation using bovine Biosheets after 6 months Bioisheets were almost completely disappeared and partial arterial reconstruction was observed. The function as a pulmonary or aortic artery was maintained for more than 6 months. Conclusion - Biosheet could function as a regenerative vascular repair material even at arteries or in xeno grafts.

Molecular cloning and characterization of a cathepsin L-like cysteine protease of Angiostrongylus cantonensis

Angiostrongylus cantonensis is a parasitic nematode dwelling in the heart and pulmonary arteries of rats, which can cause angiostrongyliasis in human by accidental infections, manifested as eosinophilic meningitis or meningoencephalitis. Cysteine proteases are the major class of endopeptidases that are expressed at a high level in A. cantonensis, which suggests it may play key roles in pathogenesis of the disease. In this study, the biological properties of the cathepsin L-like peptidase (Ac-cathL) of A. cantonensis were investigated. The Ac-cathL gene was identified from the fourth stage cDNA library of A. cantonensis, and then cloned and characterized by bioinformatics analysis and heterologous expression. The open reading frame (ORF) of Ac-cathL (1068 bp) encodes a protein of 355 amino acids with an estimated molecular weight of 58.0 kDa. Sequence analysis and multiple sequence alignment demonstrated that Ac-cathL resembles members of cathepsin L family of other parasites and mammals. Stage-dependent mRNA expression analysis showed that Ac-cathL transcripts were expressed in all stages of A. cantonensis, with the highest expression in female stage. The recombinant Ac-cathL (rAc-cathL) expressed in Escherichia coli exhibited protease activity in acidic pH as demonstrated by gelatin zymography, as well as hydrolytic activity against natural substrates, including BSA, human IgG and human fibrinogen. Immunolocalization revealed that Ac-cathL is localized in tegument of the 18 days post infection stage and uterus of the female adult stage. Therefore, these results implied that the Ac-cathL plays important roles in host tissue migration, nutrition uptake and immune evasion.

Multifunctional Triple-Layered Composite Scaffolds Combining Platelet-Rich Fibrin Promote Bone Regeneration.

There has been substantial progress made in the development of bone regeneration materials, driven by the deficiencies that exist in current clinical products, such as finite sources, donor site complications, and potential for disease transmission. To overcome these shortcomings, multifunctional scaffolds should be developed to integrate the relationship among osteoinduction, osteoconduction, and osseointegration. In this study, we fabricated polycaprolactone/gelatin (PG) nanofiber films by electrospinning, to act as barriers against connective tissue migration into bone defect sites; chitosan/poly (γ-glutamic acid)/hydroxyapatite (CPH) hydrogels were formed by electrostatic interaction and lyophilization, to exert osteoconduction; and platelet-rich fibrin (PRF) was extracted from rat abdominal aorta and combined with composite scaffolds, to promote bone induction through the release of growth factors. Hydrogels were immersed in simulated body fluid (SBF) for 1 month to investigate mineralization in vitro. Cytocompatibility, cell barrier effect, and osteogenic differentiation were also explored in vitro. The ability to effectively regenerate bone was analyzed by implantation of triple-layered composite scaffolds into rat calvarial defects in vivo. Size-matched hydrogel filled the defect site, and then, fresh PRF was applied to the hydrogel surface. Finally, P2G3 nanofiber films were applied and attached to the surrounding soft tissue. In short, we fabricated multifunctional triple-layered scaffolds by combining the advantages of osteoinduction, osteoconduction, and osseointegration, which could give full play to the role of PRF in bone regeneration and provide new and pragmatic concepts for bone tissue regeneration in clinical applications.

NCP:Fe-A Biomineral Magnetic Nanocontrast Agent for Tracking Implanted Stem Cells in Brain Using MRI.

In vivo tracking of transplanted stem cells to monitor their migration, biodistribution, and engraftment in the host tissue is important for assessing the efficacy of stem cell therapeutics. Here, we report a biomineral nanocontrast agent, iron doped calcium phosphate nanoparticles (nCP:Fe), for the in vivo tracking of stem cells in brain using magnetic resonance imaging (MRI). We have synthesized ∼100 nm sized nCP nanoparticles doped with 9.81 wt % Fe3+. In vitro studies using mesenchymal stem cells (MSCs) showed excellent biocompatibility for nCP:Fe with ∼87% labeling efficiency under optimized conditions (100 μg/mL, 6 h). Most importantly, the labeling was not found to affect the neurogenic differentiation potential of MSCs. MRI of labeled cells (∼22.34 pg Fe/cell) showed significant reduction in T2 relaxation time from 195 to 89 ms, rendering dark contrast. In vivo transplantation of labeled cells (1 × 106 cells) in external capsule of healthy rat brain showed a clearly distinguishable hypointense (dark) region in T2 weighted MR images, which remained visible up to 30 days. Subsequently, MRI tracking of labeled MSCs transplanted intracerebrally, 3 mm near to the LPS induced inflammatory site in brain, showed successful migration of labeled MSCs toward the site of inflammation. The cell migration was confirmed ex vivo by Prussian-blue (Fe3+) and Alizarin-red (Ca2+) staining of tissue sections, where individual cells were found migrated to the site of inflammation over a period of 30 days. In summary, our results clearly show that, as a biocompatible mineral composition, nCP:Fe is a promising magnetic nanocontrast agent for MRI based cell tracking in vivo.

Spatially resolved modelling of immune responses following a multiscale approach: from computational implementation to quantitative predictions

Abstract In this work we formulate a hybrid multiscale model for describing the fundamental immune processes in human immunodeficiency type 1 (HIV) infection. These include (i) the T cell migration in the lymphoid tissue, (ii) the replication cycle of HIV within an infected cell, (iii) the type I interferon (IFN) response of the target cells, and (iv) the spatiotemporal dynamics of the HIV and type I IFN fields. Computational implementation of the hybrid multiscale model is presented. It is based on the use of semi-implicit first-order symplectic Euler method for solving the equations of the second Newton’s law for cell migration and the alternating direction method for the initial-boundary value problem for reaction–diffusion equations governing the spatial evolution of the virus and IFN fields in 2D domain representing the lymph node (LN) tissue. Both, the stochastic and deterministic descriptions of the intracellular HIV infection and the IFN reaction are developed. The potential of the calibrated multiscale hybrid model is illustrated by predicting the dynamics of the local HIV infection bursts in LN tissue.

Impact of chemokine C\u2013C ligand 27, foreskin anatomy and sexually transmitted infections on HIV-1 target cell availability in adolescent South African males

We compared outer and inner foreskin tissue from adolescent males undergoing medical male circumcision to better understand signals that increase HIV target cell availability in the foreskin. We measured chemokine gene expression and the impact of sexually transmitted infections (STIs) on the density and location of T and Langerhans cells. Chemokine C–C ligand 27 (CCL27) was expressed 6.94-fold higher in the inner foreskin when compared with the outer foreskin. We show that the density of CD4+CCR5+ cells/mm2 was higher in the epithelium of the inner foreskin, regardless of STI status, in parallel with higher CCL27 gene expression. In the presence of STIs, there were higher numbers of CD4+CCR5+ cells/mm2 cells in the sub-stratum of the outer and inner foreskin with concurrently higher number of CD207+ Langerhans cells (LC) in both tissues, with the latter cells being closer to the keratin surface of the outer FS in the presence of an STI. When we tested the ability of exogenous CCL27 to induce T-cell migration in foreskin tissue, CD4 + T cells were able to relocate to the inner foreskin epithelium in response. We provide novel insight into the impact CCL27 and STIs on immune and HIV-1 target cell changes in the foreskin.

Tissue-resident memory T cells and their biological characteristics in the recurrence of inflammatory skin disorders.

The skin is the largest organ of the body. The establishment of immunological memory in the skin is a crucial component of the adaptive immune response. Once naive T cells are activated by antigen-presenting cells, a small fraction of them differentiate into precursor memory T cells. These precursor cells ultimately develop into several subsets of memory T cells, including central memory T (TCM) cells, effector memory T (TEM) cells, and tissue resident memory T (TRM) cells. TRM cells have a unique transcriptional profile, and their most striking characteristics are their long-term survival (longevity) and low migration in peripheral tissues, including the skin. Under physiological conditions, TRM cells that reside in the skin can respond rapidly to pathogenic challenges. However, there is emerging evidence to support the vital role of TRM cells in the recurrence of chronic inflammatory skin disorders, including psoriasis, vitiligo, and fixed drug eruption, under pathological or uncontrolled conditions. Clarifying and characterizing the mechanisms that are involved in skin TRM cells will help provide promising strategies for reducing the frequency and magnitude of skin inflammation recurrence. Here, we discuss recent insights into the generation, homing, retention, and survival of TRM cells and share our perspectives on the biological characteristics of TRM cells in the recurrence of inflammatory skin disorders.

EARLY OUTCOMES WITH 3D PRINTING AND AIRWAY STENTS

SESSION TITLE: Emerging Techniques for Interventional Pulmonary SESSION TYPE: Original Investigations PRESENTED ON: 10/20/2019 2:15 PM - 3:15 PM PURPOSE: Commercially-available airway stents come in limited shapes and sizes and often poorly fit in patients with complex airways. This can cause a variety of complications including, but not limited to stent occlusion, granulation tissue formation, infection, biofilm, and migration. A patient-specific stent may minimize complications, improve quality of life, and reduce the need for repeated procedures. We describe our initial clinical outcomes with the placement of thirteen patient-specific (PS) silicone stents in four patients with underlying complex airway diseases who had multiple complications associated with commercially-available airway stents METHODS: We retrospectively reviewed charts of all patients who received PS airway stents at Cleveland Clinic under FDA expanded access. Bronchoscopists completed a survey related to their experience with these PS stents, including their ascertainment of stent-related adverse events (AEs) at 24hrs and 30d post-placement RESULTS: A total of 13 patient-specific airway stents were placed in 4 patients. Two patients had Y-stents placed in the mainstem bronchi, with short limbs extending into the lobar airways. One had a Y-tracheobronchial stent, while the other had a combination of both. Twelve out of the thirteen stents placed were reported easier (30.8%) or no difference (61.5%) to load into the rigid bronchoscope, while one was reported harder (7.7%) to load. Out of the nine stents that have been removed, all nine were reported easier (22.2%) or no difference (77.8%) in difficulty to remove. At 24hrs after implantation, adverse events (AEs) were reported in 4 out of 13 stent placements. At 30 days, 7 of 13 stents were associated with AEs; all with a CTCAE (Common Terminology Criteria for Adverse Events) severity of ≤ 3. The bronchoscopists noted that this was a significant improvement in these patients compared to prior treatment with commercially-available stents. Positive or no clinical change was associated with all of the PS stents placed, and the mean duration between bronchoscopies was significantly longer with patient-specific stents than with commercially-available ones (65.6 days vs. 36.6 days, p = 0.004). Furthermore, the common AEs observed with commercially-available stents in these patients were reduced in incidence and severity, allowing for a significant increase in the lifespan of the patient-specific stents (300.2 days vs. 124.0 days, p < .001) CONCLUSIONS: Due to the increase in time to subsequent procedures and the increase in stent-life, PS silicone airway stents have the potential to decrease the total number of bronchoscopic procedures required for a patient. Overall, bronchoscopists report improved outcomes with PS stents compared to commercially-available ones, especially in patients with benign complex airway disease CLINICAL IMPLICATIONS: Patient-specific stents may have a role in patients with complications of standard stents. DISCLOSURES: No relevant relationships by Carlos Aravena Leon, source=Web Response Advisory Committee Member relationship with Intuitive Please note: $5001 - $20000 Added 11/30/2018 by Thomas Gildea, source=Web Response, value=Travel Advisory Committee Member relationship with J&J Ethicon Please note: $5001 - $20000 Added 11/30/2018 by Thomas Gildea, source=Web Response, value=Consulting fee Inventor relationship with NewCOD Please note: $1-$1000 Added 03/15/2019 by Thomas Gildea, source=Web Response, value=Non-yet Investigator relationship with Auris Please note: $5001 - $20000 Added 06/21/2019 by Thomas Gildea, source=Web Response, value=Research funding for trial Employee relationship with New COS Inc Please note: $20001 - $100000 Added 03/18/2019 by Keith Grafmeyer, source=Web Response, value=Salary No relevant relationships by Hanine Inaty, source=Web Response No relevant relationships by Michael Machuzak, source=Web Response No relevant relationships by Sonali Sethi, source=Web Response No relevant relationships by Adrianna Urbas, source=Web Response

Cadmium contamination in a soil-rice system and the associated health risk: An addressing concern caused by barium mining

Cadmium (Cd) is associated with barite; however, its biogeochemical characteristics in environments impacted by barium (Ba) mining are not known. Here, we first revealed the characteristics of Cd concentrations, distributions, and chemical forms in the soil-rice system in Ba mining areas. The associated exposure and risk assessments of Cd via rice consumption were also conducted. Elevated levels of Cd with a wide range of 0.054–91 mg/kg were found in paddy soils, approximately 63% of which exceeded the national Grade II value for soil Cd levels in China (0.3 mg/kg). A significant positive correlation between the soil Cd and soil Ba demonstrated that large amounts of Cd were released into the environment from Ba mining. Cadmium accumulated remarkably in the rice grains (0.007–3.5 mg/kg). The chemical forms in the rice plants indicated that most of the Cd was in the pectate/protein fraction (F2, 92% in the grains and 61–71% in the other tissues), followed by the residual fraction (F3, 7.1% in the grains, 27–38% in the other tissues). A minor portion of Cd was in the soluble and aminophenol fraction (F1, 0.44% in the grains, 0.26–1.4% in the other tissues). The positive correlations observed between the grain Cd and F2 in the roots, stems and leaves suggested that Cd in the rice grain was mainly from F2. Similarly, the root F2 was also positively correlated with that in the stems/leaves, indicating the critical role of F2 in Cd2+ migration in rice tissues. The estimated average hazard quotient (2.5) and annual excess lifetime cancer risk (21 × 10−5 a−1) were higher than the safety levels of 1 and 5.0 × 10−5 a−1, respectively, showing that the dietary intake of Cd via rice consumption posed high health risks to residents. Our study demonstrated that more concerns should be paid to Cd contamination in Ba mining areas.