Imunoexpressão de clic4, α-sma, e-caderina e vimentina em lesões odontogênicas epitelias benígnas / Immunoexpression of clic4, α-sma, e-cadherin and vimentin in benign epithelial odontogenic lesions

As lesões odontogênicas epiteliais benignas constituem um grupo heterogêneo de lesões. A proteína CLIC4 atua na regulação dos processos de parada de crescimento e apoptose, participando também do processo de transdiferenciação dos fibroblastos em miofibroblastos que passam a expressar α-SMA. Além disso, a expressão de CLIC4 pode interferir no processo de transição epitélio-mesenquima (TEM) em neoplasias. Este trabalho avaliou a imunoexpressão de CLIC4, α-SMA, E-caderina e Vimentina em ameloblastomas (AM) (n = 16), ceratocistos odontogênicos (n = 20) e tumores odontogênicos adenomatóides (TOA) (n = 8). A análise da expressão imunoistoquímica das proteínas CLIC4, E-caderina e vimentina no componente epitelial das lesões e de CLIC4 e α-SMA no tecido conjuntivo foi realizada de forma semi-quantitativa por um avaliador previamente calibrado. A expressão no componente epitelial de CLIC4 foi analisada separadamente no núcleo e no citoplasma, bem como a marcação de E-caderina que foi avaliada na membrana e no citoplasma. As comparações dos percentuais de imunorreatividade em relação aos grupos estudados foram realizadas por meio dos testes não paramétricos de Kruskal-Wallis e Mann-Whitney. Possíveis correlações entre a expressão de CLIC4, α-SMA, E-caderina e Vimentina foram avaliadas por meio do teste de correlação de Spearman. O nível de significância foi estabelecido em 5% (p < 0,05). Foram observados diferentes padrões de marcação entre os grupos analisados, observando-se que a imunoexpressão exclusivamente citoplasmática da CLIC4 no componente epitelial dos AM (p < 0,001) e TOA (p < 0,001) foi significativamente superior a dos CO, não demonstrarando significância estatística entre os AM e TOA. A imunoexpressão (nuclear e citoplasmática) da CLIC4 no revestimento epitelial CO foi significativamente superior à encontrada no componente epitelial dos AM (p < 0,001) e dos TOA (p < 0,001). A imunoexpressão estromal de CLIC4 foi significativamente superior nos AM (p = 0,009) e CO (p = 0,004) quando comparados aos TOA. A imunoexpressao de α-SMA significativamente maior em AM (p = 0,016) e CO (p = 0,034) quando comparados aos TOA. Para a imunoexpressão membranar da E-caderina em CO foi significativamente superior em comparação à encontrada nos AM (p = 0,009) e nos TOA (p = 0,024). Foi observada maior imunoexpressão de E-caderina (membranar e citoplasmática) nos COs, quando comparados aos AM (p < 0,001) e aos TOAs (p < 0,001). A expressão de Ecaderina citoplasmática foi significativamente maior nos AM e TOA (p < 0,001) quando comparados aos CO. Observou-se diferença estatisticamente significativa na imunoexpressão de vimentina entre os casos de AM e os casos de TOA (p = 0,038) e CO (p < 0,001), bem como entre o TOA e CO (p < 0,001). As correlações testadas entre os escores das proteínas estudadas evidenciou que no grupo dos AM foi possível evidenciar moderada correlação positiva e estatisticamente significativa (r = 0,527; p = 0,036) entre a expressão citoplasmática da CLIC4 e a expressão citoplasmática da E-caderina. Também foi verificada fraca correlação negativa e estatisticamente significativa (r = -0,499; p = 0,049) entre a expressão núcleo-citoplasmática da CLIC4 e a expressão citoplasmática da E-caderina nos AM. Além disso, uma moderada correlação positiva e estatisticamente significativa entre a expressão estromal da CLIC4 e a expressão da α-SMA nos AM (r = 0,648; p = 0,007) e nos CO (r = 0,541; p = 0,014). Foi observada forte correlação negativa e estatisticamente significativa (r = -0,813; p < 0,001) entre a expressão da E-caderina e a expressão da vimentina nos AM. Os resultados deste estudo sugerem um potencial envolvimento de CLIC4 no processo de transdiferenciação de miofibroblastos, e que a presença destas células é mais frequentemente associada a lesões de comportamento biológico mais agressivo como os AM e CO, além de uma possível atuação desta proteína na regulação do ciclo celular e na TEM nas lesões estudadas (AU).

Benign epithelial odontogenic lesions constitute a heterogeneous group of lesions. the CLIC4 protein acts in the regulation of growth arrest and apoptosis processes, also participating in the process of transdifferentiation of fibroblasts Into myofibroblasts that begin to express α-SMA. Furthermore, CLIC4 expression can interfere with the epithelialmesenchymal transition (EMT) process in neoplasms. This work evaluated the immunoexpression of CLIC4, α-SMA, e-cadherin and vimentin in ameloblastomas (AM) (n = 16), odontogenic keratocysts (OK) (n = 20) and adenomatoid odontogenic tumors (AOT) (n = 8). The analysis of the immunohistochemical expression of the proteins CLIC4, ecadherin and vimentin in the epithelial component of the lesions and of CLIC4 and α-SMA in the connective tissue was carried out in a semi-quantitative way by a previously calibrated evaluator. Expression in the epithelial component of CLIC4 was analyzed separately in the nucleus and cytoplasm, as well as e-cadherin labeling, which was evaluated in the membrane and cytoplasm. Comparisons of the percentages of immunoreactivity in relation to the studied groups were carried out using the nonparametric kruskal-wallis and mann-whitney tests. Possible correlations between the expression of CLIC4, α-SMA, e-cadherin and vimentin were evaluated using the spearman correlation test. The significance level was set at 5% (p < 0.05). Different staining patterns were observed between the groups analyzed, observing that the exclusively cytoplasmic immunoexpression of CLIC4 in the epithelial component of AM (p < 0.001) and AOT (p < 0.001) was significantly higher than that of OK, not demonstrating statistical significance between the AM and AOT. The immunoexpression (nuclear and cytoplasmic) of CLIC4 in the co epithelial lining was significantly higher than that found in the epithelial component of AM (p < 0.001) and AOT (p < 0.001). Stromal CLIC4 immunoexpression was significantly higher in AM (p = 0.009) and OK (p = 0.004) when compared to AOT. The immunoexpression of α-SMA is significantly higher in AM (p = 0.016) and OK (p = 0.034) when compared to AOT. For e-cadherin membrane immunoexpression in co was significantly higher compared to that found in AM (p = 0.009) and AOT (p = 0.024). Greater immunoexpression of e-cadherin (membrane and cytoplasmic) was observed in OK, when compared to AM (p < 0.001) and AOT (p < 0.001). Cytoplasmic ecadherin expression was significantly higher in AM and AOT (p < 0.001) when compared to OK. A statistically significant difference in vimentin immunoexpression was observed between cases of AM and cases of AOT (p = 0.038) and OK (p < 0.001), as well as between AOT and OK (p < 0.001). The correlations tested between the scores of the proteins studied showed that in the am group it was possible to demonstrate a moderate positive and statistically significant correlation (r = 0.527; p = 0.036) between the cytoplasmic expression of clic4 and the cytoplasmic expression of e-cadherin. A weak and statistically significant negative correlation (r = -0.499; p = 0.049) was also found between the nucleus-cytoplasmic expression of clic4 and the cytoplasmic expression of e- cadherin in AM. Furthermore, a moderate positive and statistically significant correlation between the stromal expression of CLIC4 and the expression of α-SMA in AM (r = 0.648; p = 0.007) and OK (r = 0.541; p = 0.014). Additionally, a strong negative and statistically significant correlation (r = -0.813; p < 0.001) was observed between the expression of ecadherin and the expression of vimentin in AM. The results of this study suggest a potential involvement of CLIC4 in the myofibroblast transdifferentiation process, and that the presence of these cells is more frequently associated with lesions with more aggressive biological behavior such as AM and OK, in addition to a possible role of this protein in the regulation of cell cycle and EMT in the lesions studied (AU).

Computational modeling and simulation of epithelial wound closure.

Wounds in the epithelium may lead to serious injurious events or chronic inflammatory diseases, however, multicellular organisms have the ability to self-repair wounds through the movement of epithelial cell toward the wound area. Despite intensive studies exploring the mechanism of wound closure, the role of mechanics in epithelial wound closure is still not well explained. In order to investigate the role of mechanical properties on wound closure process, a three-dimensional continuum physics-based computational model is presented in this study. The model takes into account the material property of the epithelial cell, intercellular interactions between neighboring cells at cell-cell junctions, and cell-substrate adhesion between epithelial cells and ECM. Through finite element simulation, it is found that the closure efficiency is related to the initial gap size and the intensity of lamellipodial protrusion. It is also shown that cells at the wound edge undergo higher stress compared with other cells in the epithelial monolayer, and the cellular normal stress dominates over the cellular shear stress. The model presented in this study can be employed as a numerical tool to unravel the mechanical principles behind the complex wound closure process. These results might have the potential to improve effective wound management and optimize the treatment.

Interleukin-17 governs hypoxic adaptation of injured epithelium.

Mammalian cells autonomously activate hypoxia-inducible transcription factors (HIFs) to ensure survival in low-oxygen environments. We report here that injury-induced hypoxia is insufficient to trigger HIF1α in damaged epithelium. Instead, multimodal single-cell and spatial transcriptomics analyses and functional studies reveal that retinoic acid-related orphan receptor γt+ (RORγt+) γδ T cell-derived interleukin-17A (IL-17A) is necessary and sufficient to activate HIF1α. Protein kinase B (AKT) and extracellular signal-regulated kinase 1/2 (ERK1/2) signaling proximal of IL-17 receptor C (IL-17RC) activates mammalian target of rapamycin (mTOR) and consequently HIF1α. The IL-17A-HIF1α axis drives glycolysis in wound front epithelia. Epithelial-specific loss of IL-17RC, HIF1α, or blockade of glycolysis derails repair. Our findings underscore the coupling of inflammatory, metabolic, and migratory programs to expedite epithelial healing and illuminate the immune cell-derived inputs in cellular adaptation to hypoxic stress during repair.

Evaluation of the alcoholic extract of Dipteryx alata Vogel almonds and bark in skin wound healing in C57BL6 mice / [Avaliação do extrato alcoólico da casca e da semente da Dipteryx alata Vogel na cicatrização da pele de camundongos C57BL/6]

The aim of this study was to evaluate the topical application of alcoholic extracts of Dipteryx alata Vogel almonds and bark in skin wound healing in mice. Fifty-four C57BL/6 mice were equally distributed into three groups: Control, Almond, and Bark. A 9 mm skin fragment was resected from the dorsal region of the animals' thorax. The wounds were submitted to topical application of base cream (vehicle), 10% hydroalcoholic almond extract, or bark extract twice a day. Macroscopic, histological, and immunohistochemical evaluations were conducted on the 7th, 14th, and 21st postoperative days. No significant difference was observed regarding skin wound area among groups, with the parameter presenting only a temporal effect on healing (p>0.05). The almond and control groups exhibited more intense collagenization than the bark group (p<0.05). Dipteryx alata Vogel showed to be inert in the wound healing process in mice.(AU)

O objetivo deste estudo foi avaliar a aplicação tópica do extrato alcoólico da semente e da casca da Dipteryx alata Vogel na cicatrização de feridas cutâneas, em camundongos. Um total de 54 camundongos C57BL/6 foram utilizados neste estudo, distribuídos em três grupos de 18 animais (controle, semente e casca). Em todos os animais, um fragmento de pele foi ressecado da região dorsal do tórax utilizando-se instrumento de punção de 9mm de diâmetro, após o qual foi realizada aplicação tópica de creme base (veículo), extrato hidroalcoólico 10% de semente ou casca, duas vezes ao dia. As avaliações macroscópica, histológica e imuno-histoquímica foram realizadas no sétimo, 14º e 21º dias de pós-operatório. Não foi observada diferença significativa quanto à área da ferida cutânea entre os grupos, apenas um efeito temporal na cicatrização (P>0,05), indicando estágio possivelmente mais avançado desse processo. Porém, na avaliação histológica, os grupos semente e controle apresentaram colagenização mais intensa que o grupo casca (P<0,05). Dipteryx alata Vogel mostrou-se inerte no processo de cicatrização de feridas em camundongos.(AU)

Modulation of the EMT/MET Process by E-Cadherin in Airway Epithelia Stress Injury.

Persistent injury and the following improper repair in bronchial epithelial cells are involved in the pathogenesis of airway inflammation and airway remodeling of asthma. E-cadherin (ECAD) has been shown to be involved in airway epithelium injury repair, but its underlying mechanisms to this process is poorly understood. Here, we describe a previously undetected function of ECAD in regulating the balance of EMT and MET during injury repair. Injury in mice and human bronchial epithelial cells (HBECs) was induced by successive ozone stress for 4 days at 30 min per day. ECAD overexpression in HBECs was induced by stable transfection. EMT features, transforming growth factor beta1 (TGF-ß1) secretion, transcriptional repressor Snail expression, and ß-catenin expression were assayed. Ozone exposure and then removal successfully induced airway epithelium injury repair during which EMT and MET occurred. The levels of TGF-ß1 secretion and Snail expression increased in EMT process and decreased in MET process. While ECAD overexpression repressed EMT features; enhanced MET features; and decreased TGF-ß1 secretion, Snail mRNA level, and ß-catenin protein expression. Moreover, activating ß-catenin blocked the effects of ECAD on EMT, MET and TGF-ß1 signaling. Our results demonstrate that ECAD regulates the balance between EMT and MET, by preventing ß-catenin to inhibit TGFß1 and its target genes, and finally facilitates airway epithelia repair.

Airway-Associated Macrophages in Homeostasis and Repair.

There is an increasing appreciation for the heterogeneity of myeloid lineages in the lung, but relatively little is known about populations specifically associated with the conducting airways. We use single-cell RNA sequencing, flow cytometry, and immunofluorescence to characterize myeloid cells of the mouse trachea during homeostasis and epithelial injury/repair. We identify submucosal macrophages, similar to lung interstitial macrophages, and intraepithelial macrophages. Following injury, there are early increases in neutrophils and submucosal macrophages, including M2-like macrophages. Intraepithelial macrophages are lost after injury and later restored by CCR2+ monocytes. We show that repair of the tracheal epithelium is impaired in Ccr2-deficient mice. Mast cells and group 2 innate lymphoid cells are sources of interleukin-13 (IL-13) that polarize macrophages and directly influence basal cell behaviors. Their proximity to the airway epithelium establishes these myeloid populations as potential therapeutic targets for airway disease.

Research progress on flat epithelium of the inner ear.

Sensorineural hearing loss and vertigo, resulting from lesions in the sensory epithelium of the inner ear, have a high incidence worldwide. The sensory epithelium of the inner ear may exhibit extreme degeneration and is transformed to flat epithelium (FE) in humans and mice with profound sensorineural hearing loss and/or vertigo. Various factors, including ototoxic drugs, noise exposure, aging, and genetic defects, can induce FE. Both hair cells and supporting cells are severely damaged in FE, and the normal cytoarchitecture of the sensory epithelium is replaced by a monolayer of very thin, flat cells of irregular contour. The pathophysiologic mechanism of FE is unclear but involves robust cell division. The cellular origin of flat cells in FE is heterogeneous; they may be transformed from supporting cells that have lost some features of supporting cells (dedifferentiation) or may have migrated from the flanking region. The epithelial-mesenchymal transition may play an important role in this process. The treatment of FE is challenging given the severe degeneration and loss of both hair cells and supporting cells. Cochlear implant or vestibular prosthesis implantation, gene therapy, and stem cell therapy show promise for the treatment of FE, although many challenges remain to be overcome.

Wound-induced polyploidization is driven by Myc and supports tissue repair in the presence of DNA damage.

Tissue repair usually requires either polyploid cell growth or cell division, but the molecular mechanism promoting polyploidy and limiting cell division remains poorly understood. Here, we find that injury to the adult Drosophila epithelium causes cells to enter the endocycle through the activation of Yorkie-dependent genes (Myc and E2f1). Myc is even sufficient to induce the endocycle in the uninjured post-mitotic epithelium. As result, epithelial cells enter S phase but mitosis is blocked by inhibition of mitotic gene expression. The mitotic cell cycle program can be activated by simultaneously expressing the Cdc25-like phosphatase String (stg), while genetically depleting APC/C E3 ligase fizzy-related (fzr). However, forcing cells to undergo mitosis is detrimental to wound repair as the adult fly epithelium accumulates DNA damage, and mitotic errors ensue when cells are forced to proliferate. In conclusion, we find that wound-induced polyploidization enables tissue repair when cell division is not a viable option.

Evolución de heridas crónicas infectadas en un paciente con elefantiasis / Evolution of Infected Chronic Wounds in a Patient with Elephantiasis

Se presenta un caso clínico de un paciente con elefantiasis y úlceras sobreinfectadas en ambas extremidades inferiores, incluyendo una valoración enfermera, diagnósticos prioritarios, plan de cuidados, justificación del tratamiento elegido y la evolución posterior. La evidencia de infección de las úlceras planteó la necesidad de instaurar una terapia con cadexómero yodado para tratar localmente la infección, e hidrogel para evitar la formación de biopelículas. Transcurrido un mes del alta del paciente se realizó un seguimiento en el que se observó una evolución favorable con aumento de tejido de granulación y tejido epitelial en varias de las lesiones presentes, por lo que se valoró de forma favorable el procedimiento de cura en ambiente húmedo; la mejora en los hábitos higiénico-dietéticos se completó por parte del paciente

We present the clinical case of a patient with elephantiasis and superinfected ulcers in both lower limbs, including the Nursing assessment, primary diagnosis, plan of care, justification for the treatment selected, and subsequent evolution. The evidence of ulcer infection required to initiate therapy with cadexomer iodine in order to treat the infection locally, and hydrogel to prevent the formation of biofilms. Follow-up was conducted one month after the patient was discharged, and favourable evolution was observed, with an increase in granulation tissue and epithelial tissue in many of the lesions; therefore, the procedure of healing in a moist environment was assessed as favourable, and the improvement in hygienic-dietary habits was completed by the patient

Repair Process Impairment by Pseudomonas aeruginosa in Epithelial Tissues: Major Features and Potential Therapeutic Avenues.

Epithelial tissues protecting organs from the environment are the first-line of defense against pathogens. Therefore, efficient repair mechanisms after injury are crucial to maintain epithelial integrity. However, these healing processes can be insufficient to restore epithelial integrity, notably in infectious conditions. Pseudomonas aeruginosa infections in cutaneous, corneal, and respiratory tract epithelia are of particular concern because they are the leading causes of hospitalizations, disabilities, and deaths worldwide. Pseudomonas aeruginosa has been shown to alter repair processes, leading to chronic wounds and infections. Because of the current increase in the incidence of multi-drug resistant isolates of P. aeruginosa, complementary approaches to decrease the negative impact of these bacteria on epithelia are urgently needed. Here, we review the recent advances in the understanding of the impact of P. aeruginosa infections on the integrity and repair mechanisms of alveolar, airway, cutaneous and corneal epithelia. Potential therapeutic avenues aimed at counteracting this deleterious impact of infection are also discussed.

In vivo evaluation of scaffolds compatible for colonoid engraftments onto injured mouse colon epithelium.

Colon organoids (colonoids) are known to be similar to colon tissue in structure and function, which makes them useful in the treatment of intestinal de-epithelialized disease. Matrigel, which is used as a transplantation scaffold for colonoids, cannot be used in clinical applications because of its undefined composition and tumorigenicity. This study identifies clinically available scaffolds that are effective for colonoid transplantation in damaged intestinal mucosa. The colon crypt was isolated and cultured from C57BL/6-Tg[CAG enhanced green fluorescent protein (EGFP)131Osb/LeySopJ mice into EGFP + colonoids and subsequently transplanted into the EDTA colitis mouse model using gelatin, collagen, or fibrin glue scaffolds. To identify scaffolds suitable for colonoid engraftment in injured colon mucosa, the success rates of transplantation and secondary EGFP colonoid formation were measured, and the scaffolds' mediated toxicity in vitro and in vivo was observed in recipient mice. When colonoids were transplanted with gelatin, collagen, and fibrin glue into the EDTA colitis mouse model, all groups were found to be successfully engrafted. Fibrin glue, especially, showed significant increase in the engrafted area compared with Matrigel after 4 wk. The scaffolds used in the study did not induce colonic toxicity after transplantation into the recipients' colons and were thus deemed safe when locally administrated. This study suggests new methods for and provides evidence of the safety and utility of the clinical application of colonoid-based therapeutics. Furthermore, the methods introduced in this study will be helpful in developing cell treatment using the esophagus or a stomach organoid for various digestive-system diseases.-Jee, J., Jeong, S. Y., Kim, H. K., Choi, S. Y., Jeong, S., Lee, J., Ko, J. S., Kim, M. S., Kwon, M.-S., Yoo, J. In vivo evaluation of scaffolds compatible for colonoid engraftments onto injured mouse colon epithelium.

Improvement of pulmonary arterial hypertension, inflammatory response, and epithelium injury by dual activation of cAMP/cGMP pathway in a rat model of monocrotaline-induced pulmonary hypertension.

Pulmonary hypertension (PH) is a life-threatening lung disease. PH with concomitant lung diseases, e.g., idiopathic pulmonary fibrosis, is associated with poor prognosis. Development of novel therapeutic vasodilators for treatment of these patients is a key imperative. We evaluated the efficacy of dual activation of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) using an active, small-molecule phosphodiesterase (PDE4)/PDE5 dual inhibitor (Compound A). Compound A increased both cAMP and cGMP levels in WI-38 lung fibroblasts and suppressed the expressions of type-1 collagen α1 chain and fibronectin. Additionally, compound A reduced right ventricular weight/left ventricular weight+septal weight ratio, brain natriuretic peptide expression levels in right ventricle, C─C motif chemokine ligand 2 expression levels in lung, and plasma surfactant protein D. Our data indicate that dual activation of cAMP/cGMP pathways may be a novel treatment strategy for PH.

Acute Colitis Drives Tolerance by Persistently Altering the Epithelial Barrier and Innate and Adaptive Immunity.

BACKGROUND: Inflammatory bowel disease (IBD) has a remitting and relapsing disease course; however, relatively little is understood regarding how inflammatory damage in acute colitis influences the microbiota, epithelial barrier, and immune function in subsequent colitis. METHODS: Mice were administered trinitrobenzene sulphonic acid (TNBS) via enema, and inflammation was assessed 2 days (d2) or 28 days (d28) later. Colitis was reactivated in some mice by re-treating at 28 days with TNBS and assessing 2 days later (d30). Epithelial responsiveness to secretagogues, microbiota composition, colonic infiltration, and immune activation was compared between all groups. RESULTS: At day 28, the distal colon had healed, mucosa was restored, and innate immune response had subsided, but colonic transepithelial transport (P = 0.048), regulatory T-cell (TREG) infiltration (P = 0.014), adherent microbiota composition (P = 0.0081), and responsiveness of stimulated innate immune bone marrow cells (P < 0.0001 for IL-1ß) differed relative to health. Two days after subsequent instillation of TNBS (d30 mice), the effects on inflammatory damage (P < 0.0001), paracellular permeability (P < 0.0001), and innate immune infiltration (P < 0.0001 for Ly6C+ Ly6G- macrophages) were reduced relative to d2 colitis. However, TREG infiltration was increased (P < 0.0001), and the responsiveness of stimulated T cells in the mesenteric lymph nodes shifted from pro-inflammatory at d2 to immune-suppressive at d30 (P < 0.0001 for IL-10). These effects were observed despite similar colonic microbiota composition and degradation of the mucosal layer between d2 and d30. CONCLUSIONS: Collectively, these results indicate that acute colitis chronically alters epithelial barrier function and both innate and adaptive immune responses. These effects reduce the consequences of a subsequent colitis event, warranting longitudinal studies in human IBD subjects.

Epithelial restitution defect in neonatal jejunum is rescued by juvenile mucosal homogenate in a pig model of intestinal ischemic injury and repair.

Intestinal ischemic injury results sloughing of the mucosal epithelium leading to host sepsis and death unless the mucosal barrier is rapidly restored. Volvulus and neonatal necrotizing enterocolitis (NEC) in infants have been associated with intestinal ischemia, sepsis and high mortality rates. We have characterized intestinal ischemia/repair using a highly translatable porcine model in which juvenile (6-8-week-old) pigs completely and efficiently restore barrier function by way of rapid epithelial restitution and tight junction re-assembly. In contrast, separate studies showed that younger neonatal (2-week-old) pigs exhibited less robust recovery of barrier function, which may model an important cause of high mortality rates in human infants with ischemic intestinal disease. Therefore, we aimed to further refine our repair model and characterize defects in neonatal barrier repair. Here we examine the defect in neonatal mucosal repair that we hypothesize is associated with hypomaturity of the epithelial and subepithelial compartments. Following jejunal ischemia in neonatal and juvenile pigs, injured mucosa was stripped from seromuscular layers and recovered ex vivo while monitoring transepithelial electrical resistance (TEER) and 3H-mannitol flux as measures of barrier function. While ischemia-injured juvenile mucosa restored TEER above control levels, reduced flux over the recovery period and showed 93±4.7% wound closure, neonates exhibited no change in TEER, increased flux, and a 11±23.3% increase in epithelial wound size. Scanning electron microscopy revealed enterocytes at the wound margins of neonates failed to assume the restituting phenotype seen in restituting enterocytes of juveniles. To attempt rescue of injured neonatal mucosa, neonatal experiments were repeated with the addition of exogenous prostaglandins during ex vivo recovery, ex vivo recovery with full thickness intestine, in vivo recovery and direct application of injured mucosal homogenate from neonates or juveniles. Neither exogenous prostaglandins, intact seromuscular intestinal layers, nor in vivo recovery enhanced TEER or restitution in ischemia-injured neonatal mucosa. However, ex vivo exogenous application of injured juvenile mucosal homogenate produced a significant increase in TEER and enhanced histological restitution to 80±4.4% epithelial coverage in injured neonatal mucosa. Thus, neonatal mucosal repair can be rescued through direct contact with the cellular and non-cellular milieu of ischemia-injured mucosa from juvenile pigs. These findings support the hypothesis that a defect in mucosal repair in neonates is due to immature repair mechanisms within the mucosal compartment. Future studies to identify and rescue specific defects in neonatal intestinal repair mechanisms will drive development of novel clinical interventions to reduce mortality in infants affected by intestinal ischemic injury.

Effects of far infrared radiation by isotropic high-density carbon on the human oral mucosa.

OBJECTIVES: Wound healing of the oral mucosal epithelium through the application of far infrared radiation emitted by isotropic high-density carbon was investigated in order to clarify the preventive and therapeutic effects of isotropic high-density carbon on oral mucosal injury. MATERIALS AND METHODS: A carbon massager with an isotropic high-density carbon tip was used. Far infrared radiation was applied to the human buccal mucosal squamous cell carcinoma cell line, HO-1-N-1 using a carbon massager, and cell growth factors and heat shock protein levels were measured using real-time RT-PCR and ELISA. Far infrared radiation was applied to oral mucosal injury in SD rats over time using the carbon massager, and its effects were examined by HE staining and immunostaining. The immunostaining positive rate was measured and analyzed using image analysis software. RESULTS: Far infrared radiation induced stronger mRNA expression and higher HSP27 and HSP70 protein levels on real-time RT-PCR and ELISA than in the control group. The far infrared radiation of oral mucosal injury in rats induced strong positive reactions, and positive rates for Ki67, HSP27, and HSP70 were higher than those in the control group. CONCLUSIONS: The treatment of oral mucosal injury with far infrared radiation emitted by isotropic high-density carbon appears to have promoted heat shock protein production and induced regenerative reactions more strongly than in the control group.

Reduction in polyamine catabolism leads to spermine-mediated airway epithelial injury and induces asthma features.

BACKGROUND: Airway epithelial injury is a crucial component of acute and severe asthma pathogenesis and a promising target for treatment of refractory asthma. However, the underlying mechanism of epithelial injury remains poorly explored. Although high levels of polyamines, mainly spermine, have been found in asthma and comorbidity, their role in airway epithelial injury and the cause of their altered levels in asthma have not been explored. METHODS: We measured key polyamine metabolic enzymes in lung samples from normal and asthmatic subjects and in mice with OVA-induced allergic airway inflammation (AAI). Polyamine metabolism was modulated using pharmacologic/genetic modulators. Epithelial stress and apoptosis were measured by TSLP levels and TUNEL assay, respectively. RESULTS: We found loss of the polyamine catabolic enzymes spermidine/spermine-N (1)-acetyltransferase-1 (SAT1) and spermine oxidase (SMOX) predominantly in bronchial epithelial cells (BECs) of human asthmatic lung samples and mice with AAI. In naïve mice, SAT1 or SMOX knockdown led to airway hyper-responsiveness, remodeling, and BEC apoptosis. Conversely, in mice with AAI, overexpression of either SAT1 or SMOX alleviated asthmatic features and reduced TSLP levels and BEC apoptosis. Similarly, while pharmacological induction of SAT1 and SMOX using the polyamine analogue bis(ethyl)norspermine (BENSPM) alleviated asthmatic features with reduced TSLP levels and BEC apoptosis, pharmacological inhibition of these enzymes using BERENIL or MDL72527, respectively, worsened them. Spermine accumulation in lungs correlated with BEC apoptosis, and spermine treatment caused apoptosis of human BEAS-2B cells in vitro. CONCLUSIONS: Spermine induces BEC injury. Induction of polyamine catabolism may represent a novel therapeutic approach for asthma via reversing BEC stress.

Occurrence of mesothelial/monocytic incidental cardiac excrescences in material from open-heart procedures: case reports and literature review

Mesothelial/monocytic incidental cardiac excrescences (MICE) are unusual findings during the histological analysis of material from the pericardium, mediastinum, or other tissues collected in open-heart surgery. Despite their somewhat worrisome histological appearance, they show a benign clinical course, and further treatment is virtually never necessary. Hence, the importance of recognizing the entity relays in its differential diagnosis, as an unaware medical pathologist may misinterpret it for a malignant neoplasm. Other mesothelial and histiocytic proliferative lesions, sharing very close histological morphology and immunohistochemistry features with MICE, have been described in sites other than the heart or the mediastinum. This similarity has led to the proposal of the common denomination "histiocytosis with raisinoid nuclei." We report three cases from the pathology archives of the Heart Institute of São Paulo University (Incor/HC-FMUSP), diagnosed as "mesothelial/monocytic incidental cardiac excrescence," with immunohistochemical documentation, and provide a literature review of this entity.

Process of tight junction recovery in the injured vocal fold epithelium: Morphological and paracellular permeability analysis.

OBJECTIVES/HYPOTHESIS: The vocal fold epithelium that includes tight junction (TJ)-based barrier function protects underlying connective tissues from external insults. TJs play an important role to control paracellular permeability of not only solutes but also ions, and preserve the vocal fold homeostasis. However, the distribution of TJs and paracellular diffusion barrier across the entire vocal fold epithelium are still unknown. The aim of this study was to identify the distribution of TJs in the vocal fold epithelium and to characterize the recovery process of TJ-based paracellular diffusion barrier in a rat model of vocal fold injury. STUDY DESIGN: Animal experiments with controls. METHODS: Normal and vocal fold-injured rats were used. Larynges were harvested for immunohistochemical examination of TJ proteins. For functional analysis, a tracer permeability assay was performed using EZ-Link Sulfo-NHS-LC-Biotin. RESULTS: TJ proteins occludin and zonula occludens 1 signals were localized to the junctional regions of the most luminal cell layers of the vocal fold epithelium. The injured region had been recovered with epithelium at 5 days postinjury, but the paracellular diffusion barrier assays revealed that biotinylation reagents diffused into the lamina propria at 5 days postinjury, and were blocked at the epithelium at 14 and 28 days postinjury. CONCLUSIONS: It was strongly suggested that TJs in the vocal fold epithelium exist at the junctional regions of the first layer of stratified squamous epithelium. TJ-based paracellular diffusion barrier following vocal fold injury is recovered by 14 days postinjury, and this period corresponds with the time course of structural changes in the regenerating epithelium layer. LEVEL OF EVIDENCE: NA. Laryngoscope, 128:E150-E156, 2018.

Lesiones epiteliales del cérvix en pacientes sin citología alterada en la Isla de la Juventud (2014-2015) / Cervix Epithelial Lesions in Patients with No Altered Cytology in the Isle of Youth (2014-2015)

Introducción: el estudio de las lesiones epiteliales del cérvix constituye una prioridad en la prevención de este cáncer que muestra un incremento evidente de la morbimortalidad, a pesar de programas de pesquiza poblacional. Objetivo: describir el comportamiento de las lesiones epiteliales según variables clínico - epidemiológicas de interés. Métodos: estudio descriptivo retrospectivo para caracterizar el comportamiento de lesiones epiteliales del cuello uterino en las 1 148 pacientes atendidas en la Consulta de Patología de Cuello en el Hospital General Docente de la Isla de la Juventud desde 2014 hasta 2015. Resultados: del total de pacientes, 71 por ciento tenían entre 25 y 51 años de edad, 44 por ciento procede de los Consejos Populares de Santa Fé y Pueblo Nuevo. El inicio de las relaciones sexuales supera los 16 años y el número promedio de partos y abortos no es mayor de dos. Del total, 42 % tenía una citología con infección por el virus del papiloma humano y 30 por ciento acudió por una lesión evidente del cérvix sin una citología alterada y 57 por ciento tenían un área lesional menor a 200 mm2. En los estudios colposcópico e histológico predominaron las lesiones de alto grado con más del 60 por ciento y las lesiones invasoras sólo representaron 4 por ciento. El número de pacientes jóvenes es significativamente alto. No se evidenció un comportamiento anormal de los factores de riesgo clásicos para esta enfermedad. Conclusiones: los resultados de este estudio pueden servir de base para trazar estrategias locales del Sistema de Salud encaminadas a la disminución de la morbimortalidad por cáncer en el territorio(AU)

Introduction: The study of cervix epithelial lesions is a priority in preventing this type of cancer because of the evident morbidity and mortality increase, despite population screening programs. Objective: To describe epithelial lesion behavior according to clinical - epidemiological variables of interest. Methods: A retrospective descriptive study was conducted to characterize the cervix epithelial lesion behavior in 1148 patients treated in the Consultation of Neck Pathology at the General Teaching Hospital of the Isle of Youth from 2014 to 2015. Results: 71 percent of the total number of patients aged between 25 and 51 years, 44 percent came from Santa Fé and Pueblo Nuevo territories. These subjects' first sexual relations were after they were 16 and the average number of deliveries and abortions is less than two. Forty-two percent had human papillomavirus infection confirmed by cytology. Thirty percent came to consultation for an obvious cervix lesion with no cytology altered. Fifty-seven percent had a lesion area less than 200 mm2. The colposcopy and histological studies, high-grade lesions predominated in more than 60 percent. Invasive lesions only represented 4 percent. The number of young patients is significantly high. There was no evidence of abnormal behavior of this disease's classic risk factors. Conclusions: The results of this study can serve as a basis to draw local strategies for the Health System aiming to reduce morbidity and mortality from cancer in the territory(AU)

Lesiones epiteliales del cérvix en pacientes sin citología alterada en la Isla de la Juventud (2014-2015) / Cervix Epithelial Lesions in Patients with No Altered Cytology in the Isle of Youth (2014-2015)

Introducción: el estudio de las lesiones epiteliales del cérvix constituye una prioridad en la prevención de este cáncer que muestra un incremento evidente de la morbimortalidad, a pesar de programas de pesquiza poblacional. Objetivo: describir el comportamiento de las lesiones epiteliales según variables clínico - epidemiológicas de interés. Métodos: estudio descriptivo retrospectivo para caracterizar el comportamiento de lesiones epiteliales del cuello uterino en las 1 148 pacientes atendidas en la Consulta de Patología de Cuello en el Hospital General Docente de la Isla de la Juventud desde 2014 hasta 2015. Resultados: del total de pacientes, 71 por ciento tenían entre 25 y 51 años de edad, 44 por ciento procede de los Consejos Populares de Santa Fé y Pueblo Nuevo. El inicio de las relaciones sexuales supera los 16 años y el número promedio de partos y abortos no es mayor de dos. Del total, 42 % tenía una citología con infección por el virus del papiloma humano y 30 por ciento acudió por una lesión evidente del cérvix sin una citología alterada y 57 por ciento tenían un área lesional menor a 200 mm2. En los estudios colposcópico e histológico predominaron las lesiones de alto grado con más del 60 por ciento y las lesiones invasoras sólo representaron 4 por ciento. El número de pacientes jóvenes es significativamente alto. No se evidenció un comportamiento anormal de los factores de riesgo clásicos para esta enfermedad. Conclusiones: los resultados de este estudio pueden servir de base para trazar estrategias locales del Sistema de Salud encaminadas a la disminución de la morbimortalidad por cáncer en el territorio(AU)

Introduction: The study of cervix epithelial lesions is a priority in preventing this type of cancer because of the evident morbidity and mortality increase, despite population screening programs. Objective: To describe epithelial lesion behavior according to clinical - epidemiological variables of interest. Methods: A retrospective descriptive study was conducted to characterize the cervix epithelial lesion behavior in 1148 patients treated in the Consultation of Neck Pathology at the General Teaching Hospital of the Isle of Youth from 2014 to 2015. Results: 71 percent of the total number of patients aged between 25 and 51 years, 44 percent came from Santa Fé and Pueblo Nuevo territories. These subjects' first sexual relations were after they were 16 and the average number of deliveries and abortions is less than two. Forty-two percent had human papillomavirus infection confirmed by cytology. Thirty percent came to consultation for an obvious cervix lesion with no cytology altered. Fifty-seven percent had a lesion area less than 200 mm2. The colposcopy and histological studies, high-grade lesions predominated in more than 60 percent. Invasive lesions only represented 4 percent. The number of young patients is significantly high. There was no evidence of abnormal behavior of this disease's classic risk factors. Conclusions: The results of this study can serve as a basis to draw local strategies for the Health System aiming to reduce morbidity and mortality from cancer in the territory(AU)