JNK inhibition enhances cell-cell adhesion impaired by desmoglein 3 gene disruption in keratinocytes.

c-Jun NH2-terminal protein kinase (JNK) and p38 are stress-activated mitogen-activated protein kinases (MAPK) that are phosphorylated by various stimuli. It has been reported that the loss of desmoglein (DSG) 3, a desmosomal transmembrane core molecule, in keratinocytes impairs cell-cell adhesion accompanied by p38 MAPK activation. To understand the biological role of DSG3 in desmosomes and its relationship with stress-activated MAPKs, we established DSG3 knockout keratinocytes (KO cells). Wild-type cells showed a linear localization of DSG1 to cell-cell contacts, whereas KO cells showed a remarkable reduction despite the increased protein levels of DSG1. Cell-cell adhesion in KO cells was impaired over time, as demonstrated by dispase-based dissociation assays. The linear localization of DSG1 to cell-cell contacts and the strength of cell-cell adhesion were promoted by the pharmacological inhibition of JNK. Conversely, pharmacological activation of JNK, but not p38 MAPK, in wild-type cells reduced the linear localization of DSG1 in cell-cell contacts. Our data indicate that DSG1 and DSG2 in KO cells cannot compensate for the attenuation of cell-cell adhesion strength caused by DSG3 deficiency and that JNK inhibition restores the strength of cell-cell adhesion by increasing the linear localization of DSG1 in cell-cell contacts in KO cells. Inhibition of JNK signaling may improve cell-cell adhesion in diseases in which DSG3 expression is impaired.

A patient with pleuroparenchymal fibroelastosis carrying a novel fibrillin-2 gene variant.

Pleuroparenchymal fibroelastosis is a recently recognized clinical entity characterized by interstitial pneumonia with proliferating elastin in the upper lung regions. Pleuroparenchymal fibroelastosis is categorized as idiopathic or reported depending on the coexistent initiating factors; however, congenital contractural arachnodactyly, which is caused by abnormal production of elastin based on a mutation in the fibrillin-2 gene, is rarely reported with lung lesion resembling pleuroparenchymal fibroelastosis. We present a case of pleuroparenchymal fibroelastosis in a patient with a novel mutation in the fibrillin-2 gene, which encodes the prenatal fibrillin-2 protein as a scaffold for elastin.

Inhibition of retinoid X receptor improved the morphology, localization of desmosomal proteins and paracellular permeability in three-dimensional cultures of mouse keratinocytes.

Retinoic acid (RA) plays an important role in epithelial homeostasis and influences the morphology, proliferation, differentiation and permeability of epithelial cells. Mouse keratinocytes, K38, reconstituted non-keratinized stratified epithelium in three-dimensional (3D) cultures with serum, which contains retinol (a source of RA), but the morphology was different from in vivo epithelium. The formed epithelium was thick, with loosened cell-cell contacts. Here, we investigated whether the inhibition of RA receptor (RAR)/retinoid X receptor (RXR)-mediated signaling by an RXR antagonist, HX 531, improved K38 3D cultures in terms of morphology and intercellular junctions. The epithelium formed by 0.5 µM HX531 was thin, and the intercellular space was narrowed because of the restoration of the layer-specific distribution of desmoglein (DSG)-1, DSG3 and plakoglobin (PG). Moreover, the levels of desmosomal proteins and tight junction proteins, including DSG1, DSG2, DSG3, PG, claudin (CLDN)-1 and CLDN4 increased, but the adherens junction protein, E-cadherin, did not show any change. Furthermore, CLDN1 was recruited to occludin-positive cell-cell contacts in the superficial cells and transepithelial electrical resistance was increased. Therefore, K38 3D cultures treated with 0.5 µM HX531 provides a useful in vitro model to study intercellular junctions in the non-keratinized epithelium.

Factors That Influence the Judgment of Oral Management Necessity in Preoperative Oral Screening.

Oral management during the perioperative period is important to prevent the development of postoperative complications. However, there are no unified systems to examine the oral status of patients and very few studies have focused on preoperative oral screening. In this study, we examined the oral status of patients who underwent oral screening at a University Hospital. A total of 1173 patients who underwent oral screening for perioperative management from April 2020 to July 2021 were enrolled. The subjects' medical data were retrospectively extracted from the dental records, and finally, the data of 1081 patients aged ≥20 years were analyzed. Oral screening based on seven categories was performed by dentists or dental hygienists. Our cumulative results determined whether patients required oral management during the perioperative period. "Poor oral hygiene" was the most frequent category (24%) of all oral categories examined. Logistic analysis revealed that tooth mobility had the highest odds ratio (21.476; 95% confidence interval: 11.462-40.239; p < 0.001) for oral management necessity during the perioperative period. Our study suggests that poor oral hygiene is most frequently observed in preoperative oral screening. Moreover, tooth mobility in preoperative oral screening may influence the judgment of oral management necessity during the perioperative period.

Effect of Mucosal Brushing on the Serum Levels of C-Reactive Protein for Patients Hospitalized with Acute Symptoms.

This study was based in a hospital setting. Patients with acute symptoms face a life-threatening crisis and often have systemic complications during the convalescence stage. During the acute stage, oral function does not work and oral hygiene status deteriorates. A gauze or sponge brush is generally used to wipe the oral cavity; however, this process does not clean the oral cavity enough. Effective oral care requires better methods. Patients participating in this study were all hospitalized by ambulance and with acute symptoms. During the convalescence stage, patients were assigned application of mucosal brushing or wiping by gauze or sponge brush by order of hospitalization. The effects were evaluated by the number of bacteria on the tongue surface, serum C-reactive protein (CRP) and body temperature. Changes in bacterial count, body temperature, and CRP were effectively reduced in the mucosal brushing group compared to the wiping by gauze or sponge brush group. Based on mixed effect modeling, the coefficient of mucosal brushing for CRP was -2.296 and for body temperature was -0.067 and statistically significant. This simple method can effectively prevent systemic complication of inpatients with deteriorated oral conditions. This method may also be effective for the elderly in nursing homes or perioperative oral-care management.

The reduced susceptibility of mouse keratinocytes to retinoic acid may be involved in the keratinization of oral and esophageal mucosal epithelium.

Keratinocytes take up serum-derived retinol (vitamin A) and metabolize it to all-trans-retinoic acid (atRA), which binds to the nuclear retinoic acid receptor (RAR). We previously reported that serum-affected keratinocyte differentiation and function; namely, it inhibited keratinization, decreased loricrin (LOR) and claudin (CLDN) 1 expression, increased keratin (K) 4 and CLDN4 levels, and reduced paracellular permeability in three-dimensional (3D) cultures of mouse keratinocytes (COCA). Contrarily, RAR inhibition reversed these changes. Here, we aimed to examine whether atRA exerted the same effects as serum, and whether it was involved in the differential oral mucosa keratinization among animal species. Porcine oral mucosal keratinocytes, which form non-keratinized epithelium in vivo, established keratinized epithelium in 3D cultures. Both mouse and porcine sera induced non-keratinized epithelium at 0.1% in COCA 3D cultures. Although atRA caused the same changes as serum, its effective concentration differed. atRA inhibited keratinization at 0.1 nM and 1 nM in porcine or human keratinocytes and COCA, respectively. Furthermore, atRA upregulated CLDN7 in the cytoplasm but not in cell-cell contacts. These atRA-induced changes were reverted by RAR inhibition. The results indicate that serum-induced changes are probably due to the effect of serum-derived atRA, and that mouse keratinocytes require higher atRA concentrations to suppress keratinization than porcine and human keratinocytes. We propose that the lower susceptibility of mouse keratinocytes to atRA, rather than a lower retinol concentration, is a possible reason for the keratinization of mouse oral mucosal epithelium.

Risk factors for postoperative pneumonia according to examination findings before surgery under general anesthesia.

OBJECTIVE: This study was performed to determine the risk factors associated with postoperative complications after surgery under general anesthesia according to respiratory function test results and oral conditions. MATERIALS AND METHODS: Preoperative examination data were collected for 471 patients who underwent surgery under general anesthesia at the Medical Hospital of Kyusyu University. Respiratory function tests, oral examinations, and perioperative oral management were performed in all patients. The incidence of and risk factors for postoperative complications were investigated. Classification and regression tree analyses were performed to investigate the risk factors for postoperative complications. RESULTS: Among the 471 patients, 11 developed postoperative pneumonia, 10 developed postoperative respiratory symptoms, and 10 developed postoperative fever. The most important risk factor for pneumonia was edentulism. Age, the Brinkman index, and head and neck surgery were also revealed as important risk factors for pneumonia. The O'Leary plaque control record (initial visit) was an important risk factor for postoperative respiratory symptoms. With respect to postoperative fever, a Hugh-Jones classification of grade > 1 was the most important risk factor; edentulism and a Brinkman index of > 642.5 were also found to be risk factors. CONCLUSION: In addition to respiratory function tests, oral examinations may be important for the prediction of postoperative complications. Additionally, improved oral hygiene may be effective in preventing postoperative respiratory complications. CLINICAL RELEVANCE: Risk factors for postoperative complications should be comprehensively evaluated using both respiratory function tests and oral findings.

Inhibition of JNK in HaCaT cells induced tight junction formation with decreased expression of cytokeratin 5, cytokeratin 17 and desmoglein 3.

Epidermal keratinocytes proliferate in the basal layer, differentiate, migrate through the spinous layer, granular layer and cornified layer, and finally are peeled off from the surface of skin with layer-specific expression of differentiation markers, including cytokeratins and cell-cell junction proteins such as desmogleins. Basal cells express CK5, CK14 and Ki67. In contrast, the suprabasal cells in the spinous and granular layers express CK1 and CK10 without Ki67. Inhibition of c-Jun NH2-terminal protein kinase (JNK) in HaCaT cells, a human epidermal keratinocyte cell line, induced the formation of tight junctions, which occurs in the granular layer in vivo. These cells lost their expression of CK5 and CK17, exhibited decreased expression of desmoglein 3 and had no Ki67 labeling in the nucleus. These results suggest that inhibition of JNK causes HaCaT cells to differentiate from basal- and spinous-like cells to granular-like cells. The inhibition of JNK in HaCaT cells provides a useful in vitro model system to study the differentiation of epidermal keratinocytes.

Protective effects of topical application of a poorly soluble antioxidant astaxanthin liposomal formulation on ultraviolet-induced skin damage.

Astaxanthin (Asx) would be expected to prevent ultraviolet (UV)-induced skin damage, as it is regarded as a potent antioxidative carotenoid in biological membranes. However, it is difficult to administer Asx topically to skin because of its poor water solubility. In this study, we attempted to solve this problem by preparing liposomes containing Asx (Asx-lipo), which were dispersible in the water phase, and therefore, suitable for topical application to the skin. Asx-lipo was shown to have potent scavenging ability against chemiluminescence-dependent singlet oxygen production in the water phase. When Asx-lipo was applied to skin before UV exposure, UV-induced skin thickening was prevented. Interestingly, collagen reduction induced by UV exposure was also prevented by preadministration of Asx-lipo. In addition, topical administration of Asx-lipo containing cationic lipid inhibited melanin production in skin exposed to UV. Consequently, we succeeded in preventing UV-induced skin damage using a topical application of a liposomal formulation containing Asx.