Inflammation modulates intercellular adhesion and mechanotransduction in human epidermis via ROCK2.

Aberrant mechanotransduction and compromised epithelial barrier function are associated with numerous human pathologies including inflammatory skin disorders. However, the cytoskeletal mechanisms regulating inflammatory responses in the epidermis are not well understood. Here we addressed this question by inducing a psoriatic phenotype in human keratinocytes and reconstructed human epidermis using a cytokine stimulation model. We show that the inflammation upregulates the Rho-myosin II pathway and destabilizes adherens junctions (AJs) promoting YAP nuclear entry. The integrity of cell-cell adhesion but not the myosin II contractility per se is the determinative factor for the YAP regulation in epidermal keratinocytes. The inflammation-induced disruption of AJs, increased paracellular permeability, and YAP nuclear translocation are regulated by ROCK2, independently from myosin II activation. Using a specific inhibitor KD025, we show that ROCK2 executes its effects via cytoskeletal and transcription-dependent mechanisms to shape the inflammatory response in the epidermis.

IL-38 orchestrates proliferation and differentiation in human keratinocytes.

Interleukin (IL)-38 is a member of the IL-1 cytokine family with reported anti-inflammatory activity. The highest constitutive IL-38 expression is detected in the skin, where it is mainly produced by differentiating keratinocytes. However, little data are available regarding its biological functions. In this study, we investigated the role of IL-38 in skin physiology. We demonstrate here that dermal fibroblasts and epithelial cells of skin appendages, such as eccrine sweat glands and sebaceous glands, also express IL-38. Next, using two- and three-dimensional cell cultures, we show that endogenous expression of IL-38 correlates with keratinocyte differentiation and its ectopic overexpression inhibits keratinocyte proliferation and enhances differentiation. Accordingly, immunohistochemical analysis revealed downregulation of IL-38 in skin pathologies characterized by keratinocyte hyperproliferation, such as psoriasis and basal or squamous cell carcinoma. Finally, intracellular IL-38 can shuttle between the nucleus and the cytoplasm and its overexpression modulates the activity of the transcription regulators YAP and ID1. Our results indicate that IL-38 can act independently from immune system activation and suggest that it may affect the epidermis directly by decreasing proliferation and promoting differentiation of keratinocytes. These data suggest an important role of keratinocyte-derived IL-38 in skin homeostasis and pathologies characterized by epidermal alterations.

Sexual function and health status in adult patients with Congenital Adrenal Hyperplasia.

INTRODUCTION: Congenital Adrenal Hyperplasia (CAH) is the most common reason for undifferentiated genital appearance in new-borns. Psychosexual outcome in women with CAH has been rarely evaluated, but it seems to be one of the most important factors determining the indications for the surgical treatment of CAH. OBJECTIVE: This is to assess sexual function and the health status (HS) in adult females with CAH who had feminizing genitoplasty in childhood. MATERIAL AND METHOD: The protocol was approved by the Ethical Committee, and the hospital database was searched for patients with CAH who had genitoplasty between 1975 and 2000. 57 adult patients were identified, and 9 (18%) patients agreed to participate in the study. Mean age at operation was 5.4 years, and mean follow-up duration was 10.9 years. The Female Sexual Function Index (FSFI) was used to evaluate sexual function, and the 36-item Short Form Health Survey (SF-36) was used to evaluate their health status (HS). A FSFI score < 26,55 was classified as Female Sexual Dysfunction (FSD). The control group consisted of 10 adult female volunteers of comparable age, without any oncological or chronic diseases. Fisher's exact test was used for statistical analysis. RESULTS: All patients in the CAH group had female gender identity. One was homosexual, and one reported not having any sexual activity. In the control group, all patients had female gender identity. All were heterosexual and one reported not having any sexual activity. The sexual function in five domains and total score were similar in both groups. More pain was reported in the CAH group as compared with the control group, and it was statistically significant. In the CAH group, 5/9 patients had FSD. In the control group, 4/10 patients had FSD. The difference was statistically insignificant (p = 0.66). Mean SF-36 score in the CAH group was 47.1 points, while it was 46.7 points in the control group. The testosterone level in all CAH patients was within the normal range (0.13-1.1 ng/ml). The 17-OH progesterone level was above normal range in 5/9 (55.6%) patients with CAH. All women in the CAH group were hormonally treated. In the control group, all patients had a normal testosterone level (0.15-0.68 ng/ml); the 17-OH progesterone level was in normal range in this group. DISCUSSION: We compared our results with the literature data, which used the same questionnaires as in our study. CONCLUSIONS: Health status and sexual function in the traceable CAH patients didn't differ from the control group.

IL-17E (IL-25) and IL-17A Differentially Affect the Functions of Human Keratinocytes.

Our group has recently shown that keratinocyte-derived IL-17E (IL-25), one of six members of the IL-17 family, is overexpressed in lesional psoriatic skin and is involved in its pathophysiology. We show here that IL-22 enhances IL-17E production in human keratinocytes and that these cells display a complete IL-17E receptor at their surface, the expression of which is further induced by IL-17A, indicating a potential autocrine effect of IL-17E. Therefore, we addressed the impact of IL-17E on the function of human primary keratinocytes. IL-17E promoted the proliferation of keratinocytes in two-dimensional and three-dimensional cultures and caused the concomitant upregulation of differentiation-associated gene transcripts (e.g., keratin 10), whereas their expression was either inhibited or not changed by IL-17A. Contrary to IL-17A, IL-17E was not involved in the induction of antimicrobial proteins. Time-lapse analysis of cell movement showed that IL-17E influences cell motility, increasing both cell speed and displacement. This was associated with specific changes in the actin cytoskeleton organization and the cell-substrate adhesion. No such effects were observed upon IL-17A stimulation. In summary, we identified effects of IL-17E clearly distinct from IL-17A, pointing toward an important role of IL-17E in the physiology and pathophysiology of the epidermis.

Interleukin-38 interacts with destrin/actin-depolymerizing factor in human keratinocytes.

Interleukin (IL)-38 is a member of the IL-1 family of cytokines, which was proposed to exert anti-inflammatory effects. IL-38 is constitutively expressed in the skin, where keratinocytes are the main producing cells. Little information is currently available concerning IL-38 biology. Here, we investigated the subcellular localization and interaction partners of the IL-38 protein in human keratinocytes. IL-38 expression was reduced in primary keratinocytes grown in monolayer (2D) cultures. We thus used IL-38 overexpressing immortalized normal human keratinocytes (NHK/38) to study this cytokine in cell monolayers. In parallel, differentiation of primary human keratinocytes in an in vitro reconstructed human epidermis (RHE) 3D model allowed us to restore endogenous IL-38 expression. In NHK/38 cells and in RHE, IL-38 was mainly cell-associated, rather than released into culture supernatants. Intracellular IL-38 was preferentially, although not exclusively, cytoplasmic. Similarly, in normal human skin sections, IL-38 was predominantly cytoplasmic in the epidermis and essentially excluded from keratinocyte nuclei. A yeast two-hybrid screen identified destrin/actin-depolymerizing factor (DSTN) as a potential IL-38-interacting molecule. Co-immunoprecipitation and proximity ligation assay confirmed this interaction. We further observed partial co-localization of IL-38 and DSTN in NHK/38 cells. Endogenous IL-38 and DSTN were also co-expressed in all epidermal layers in RHE and in normal human skin. Finally, IL-38 partially co-localized with F-actin in NHK/38 cells, in particular along the cortical actin network and in filopodia. In conclusion, IL-38 is found predominantly in the cytoplasm of human keratinocytes, where it interacts with DSTN. The functional relevance of this interaction remains to be investigated.

Relation between ureteral telocytes and the hydronephrosis development in children.

INTRODUCTION: Hydronephrosis is an actual pediatric problem, affecting children in the ante- and neonatal periods. Intrinsic stenosis is due to external obstruction and creates a pathophysiological basis of this urological pathology. Co-localization of ureter with a renal vasculature also could not be omitted from this point of view. Mesenchymal cells, partially telocytes, are important for local fibrosis development and hydronephrosis formation as well. In the current study, we focused on identification of telocytes in the human ureters to hypothesize their role in hydronephrosis pathophysiology. MATERIAL AND METHODS: the samples were taken from 18 surgically treated patients with hydronephrosis (due to ureteral obstruction and crossing renal vessel). The control group consisted of 10 patients suffered from a non-obstructive disease of the urinary tract - predominantly renal tumors. Tissue samples from a ureter were stained for c-kit, tryptase, CD34 and PDGFRα to identify telocytes. Routine histology was performed to analyze tissue morphology, collagen deposits and mast cell's profile. RESULTS: Telocytes were detected in the ureteral wall. In patients with hydronephrosis we revealed decreasing density of telocytes, the prevalence of collagen, rise in mast cells amount and the ureteral wall thickening. In ureters with crossing renal vessels as a primary etiologic factor more telocytes have been observed in comparison with the obstructive hydronephrosis. CONCLUSIONS: A declined density of telocytes accompanied hydronephrosis development. Increased number of mast cells in the ureteral wall reflects a local inflammation, while detailed observation of collagen/muscle deposits and density of telocytes reveal a difference depended on etiologic factor (obstruction or crossing vessel) in patients with hydronephrosis.

Can We Further Improve the Quality of Nephro-Urological Care in Children with Myelomeningocele?

Myelomeningocele (MMC) results from a failure of normal neural tube fusion in early fetal development. Retrospective, observational study of medical data of 54 children treated in Pediatric Nephrology and Urology Clinics for five years was performed. The following data were analyzed: serum creatinine, eGFR, urine analysis, renal scintigraphy (RS), renal ultrasound, and urodynamics. Mean age of studied population: 12.3 years, median of eGFR at the beginning and at the end of survey was 110.25 and 116.5 mL/min/1.73 m² accordingly. Median of frequency of urinary tract infections (fUTI): 1.2 episodes/year. In 24 children: low-pressure, in 30 children: high-pressure bladder was noted. Vesicouretral reflux (VUR) was noted in 23 children (42.6%). fUTI were more common in high-grade VUR group. High-grade VURs were more common in group of patients with severe renal damage. At the end of the survey 11.1% children were qualified to higher stages of chronic kidney disease. Renal parenchyma damage progression in RS was noted in 22.2% children. Positive VUR history, febrile recurrent UTIs, bladder wall trabeculation, and older age of the patients constitute risk factors of abnormal renal scans. More than 2.0 febrile, symptomatic UTIs annually increase by 5.6-fold the risk of severe renal parenchyma damage after five years.

Cosmetic Outcomes of Sutureless Closure in Gastroschisis.

Purpose A sutureless gastroschisis repair allows for spontaneous closure of abdominal wall defect. We report our experience focusing on final esthetic outcome. Methods Retrospective data were collected from medical reports of all neonates with gastroschisis operated from January 2009 to December 2013. Variables recorded included patients descriptors, management modality, and cosmetic outcome. Results From the overall group of 38 patients with gastroschisis, 20 infants treated with sutureless closure were included in this study. In the analyzed cohort, 17 (85%) children were operated under general anesthesia and 3 (15%) without intubation. Primary reduction was possible in 15 (75%) cases, and in 5 (25%) we used silo. There were two (10%) deaths in late postoperative course due to septic complications. Three (15%) infants needed laparotomy because of adhesions and bowel obstruction. There were no infectious complications of the wound. Only 55% (10/18) of children presented umbilical hernia prior to discharge. Only two (11%) children with umbilical hernia were operated until now. Almost all patients (16/18; 89%) present excellent final cosmetic result without scar formation. Conclusion Sutureless closure of uncomplicated gastroschisis is a safe technique that reduces need of intubation and provides excellent cosmetic results.

Interstitial cells of Cajal and telocytes in the urinary system: facts and distribution.

Current knowledge confirms the existence of interstitial cells of Cajal (ICCs) and telocytes in the urinary system (kidneys, ureter and urinary bladder). Therefore, summarizing of available data can be helpful in understanding of pathophysiology of urological disorders. Telocytes (TCs) are a newly discovered type of cell with numerous functions, described in vertebrates (fish, reptiles, birds, mammals, including human). Despite unique characteristics, they have own differences in morphology and properties in urinary bladder and other organs of the urinary system. This review summarizes particular features of ICCs and TCs in the urinary system, emphasizing their involvement in physiological and pathophysiological processes of the urinary bladder.

Results of varicocele treatment with laparoscopic Palomo technique in children and adolescents in years 2002-2012.

INTRODUCTION: Varicocele is caused by enlargement of internal testicular veins, including the pampiniform veno- us plexus. It is assumed that it occurs in 8% to 16% of the male population and predisposes to male infertility. The aim of our study was to assess laparoscopic treatment of the varicocele in children using the Palomo method. MATERIAL AND METHODS: In the years 2002-2012, our clinic treated 114 boys with the varicocele on the left side. In all patients the procedure involved ligation of veins, the testicular artery and lymphatic vessels with the Roeder knot. The average time of the procedure was 29 min. RESULTS: Varicocele recurrence was found in 2 out of 94 patients reporting for the follow-up visit, and this corresponds to 2.1% of the studied group. The postoperative hydrocele occurred in 20 patients, of which 5 (5.3% of the studied group) required surgical treatment with the Winkelmann technique, while in 15 (15.9% of the studied group) disappeared spontaneously during the average follow-up period of 1 year. No other intra- or postoperative complications occurred in any patient. Also no case of testicular atrophy was diagnosed. CONCLUSION: The results obtained from our laparoscopy patients using the Palomo method in treatment of varicocele can be considered satisfactory.

Pediatric single port transumbilical nephrectomy and nephroureterectomy.

OBJECTIVE: To present seven cases of single incision laparoscopic nephrectomy and nephroureterectomy in children as a recent videoscopic innovation. PATIENTS AND METHODS: Seven children with nonfunctioning kidneys, three with multicystic dysplastic kidneys, two with end-stage renal nephropathy due to vesicoureteral reflux, and two with giant hydronephrosis were qualified to nephrectomy or nephroureterectomy. The surgery was performed transperitoneally using single incision access laparoscopy. The operative time was in the range of 50-90 min. RESULTS: There were no intraoperative or postoperative complications. The patients were discharged on the third postoperative day. The incision scars were hidden inside the umbilicus. CONCLUSIONS: Nephrectomy or nephroureterectomy using a single transumbilical port in children is a feasible and efficacious technique. The advantages are shortened convalescence, excellent cosmetic results, and reduction of potential wounds complications. However, clear indication of single site laparoscopic procedures in children remains to be clarified.