Echinococcus granulosus cyst fluid suppresses inflammatory responses by inhibiting TRAF6 signalling in macrophages.

Echinococcus granulosus sensu lato has complex defence mechanisms that protect it from the anti-parasitic immune response for long periods. Echinococcus granulosus cyst fluid (EgCF) is involved in the immune escape. Nevertheless, whether and how EgCF modulates the inflammatory response in macrophages remains poorly understood. Here, real-time polymerase chain reaction and enzyme-linked immunosorbent assay revealed that EgCF could markedly attenuate the lipopolysaccharide (LPS)-induced production of pro-inflammatory factors including tumour necrosis factor-α, interleukin (IL)-12 and IL-6 but increase the expression of IL-10 at mRNA and protein levels in mouse peritoneal macrophages and RAW 264.7 cells. Mechanically, western blotting and immunofluorescence assay showed that EgCF abolished the activation of nuclear factor (NF)-κB p65, p38 mitogen-activated protein kinase (MAPK) and ERK1/2 signalling pathways by LPS stimulation in mouse macrophages. EgCF's anti-inflammatory role was at least partly contributed by promoting proteasomal degradation of the critical adaptor TRAF6. Moreover, the EgCF-promoted anti-inflammatory response and TRAF6 proteasomal degradation were conserved in human THP-1 macrophages. These findings collectively reveal a novel mechanism by which EgCF suppresses inflammatory responses by inhibiting TRAF6 and the downstream activation of NF-κB and MAPK signalling in both human and mouse macrophages, providing new insights into the molecular mechanisms underlying the E. granulosus-induced immune evasion.

Value of cytopathologist review of ultrasound examinations in non-diagnostic/unsatisfactory thyroid FNA.

BACKGROUND: Correlation of cytologic and ultrasound findings is extremely valuable for the cytopathologist in management of thyroid nodules. METHODS: Ultrasound scans (US) of all thyroid FNA taken over a 13 month period and reported by a single cytologist were reviewed at the time of reporting, focusing on aspirates that were non-diagnostic/unsatisfactory, equivalent to Bethesda Class I, UK Royal College of Pathologists Class Thy1 or Thy1c. RESULTS: FNA cases [68 (40.7%)] were classified as Thy1, equivalent to Bethesda Class I. US of 3 Thy1 cases were not available for review. On cytologist US review 9 cases were classified as pure cystic, 28 as mixed cystic/solid, 12 as predominantly solid/focally cystic and 16 as purely solid. 27 (41.5%) of cases on cytological assessment were Thy1 and showed no evidence of a cyst on US, 17 (26.1%) were Thy1/Thy1c showing features suggestive of a possible cyst and 21 (32.3%) were Thy1c showing definite features of a cyst. Fifteen of 16 (93.7%) of pure solid cases on US were Thy1, equivalent to Bethesda Class I and all 9 (100%) of cases that were pure cystic on US were reported as Thy1c-equivalent to Bethesda Category I-cyst fluid only (P < .001). CONCLUSION: Cytopathologist review of thyroid US is extremely useful and can be helpful in triaging patients for further management in cases of solid, mixed cystic and/or solid, and pure cystic thyroid lesions with non-diagnostic/unsatisfactory thyroid FNA.

Malignancy Rates of Thyroid Cytology: Cyst Fluid Benign or Non-Diagnostic?

BACKGROUND We sought to investigate subgroup distribution using Bethesda classification and risks for malignancy. We also compared the malignancy risk of cases that were denoted as non-diagnostic due to cystic contents, with cases that were denoted as non-diagnostic due to presence of other features. MATERIAL AND METHODS The study included pathology test results of 1,440 thyroid nodule samples diagnosed using Bethesda classification. Results of 305 thyroidectomy excision specimens from these patients were also compared with cytology results to determine the frequency of malignancy. The non-diagnostic group was divided into two categories: those with cystic contents, and others. Malignancy rates were separately calculated for the two groups, and compared with the other classification groups. RESULTS Distribution of malignancy rates by Bethesda classification were as follows: non-diagnostic 12.5% (6/48), benign 1.5% (3/198), atypia of undetermined significance/follicular lesion of undetermined significance (AFLUS) 9% (1/11), suspicious for follicular neoplasm (SFN) 37.5% (3/8), suspicious malignancy 70% (8/26), malignancy 100% (14/14). CONCLUSIONS Despite the limited number of cases, our study concluded that cystic content was closer to the benign category than the non-diagnostic category if the assessment was based on malignancy rates. In this group, similar to aspirations containing plenty of lymphocytes that indicates colloid or lymphocytic thyroiditis, it is still controversial whether criterion for adequacy of follicular epithelial cells should be sought, or if they should be regarded as benign in order to prevent unnecessarily performance of repeat aspirations.

Hydatid cyst fluid promotes peri-cystic fibrosis in cystic echinococcosis by suppressing miR-19 expression.

BACKGROUND: Echinococcus granulosus infection causes cystic echinococcosis (CE); the generation of liver fibrosis around the parasitic larval cyst (metacestode) may play a major role in the spontaneous limitation of the parasitic growth; however, fibrogenesis has received little attention in CE. It has been reported that miR-19b plays a role in various diseases, including infectious diseases, by regulating fibrogenesis. However, its function in the development of liver fibrosis in E. granulosus infection is unknown. METHODS: The expression of miR-19b and genes that are involved in liver fibrosis were analysed in E. granulosus-infected human livers using qRT-PCR. The role of miR-19b on hepatic stellate cells (LX-2 cells in vitro) treated with hydatid cyst fluid (HCF) was then analysed by 3-(4, 5-dimet-hylthiazol-2-yl)-2, 4-diphenyl-tetrazolium bromide (MTT) assay, qRT-PCR, Western blot and flow cytometry. RESULTS: The results showed that the expression of miR-19 was significantly reduced in the pericystic collagen-rich liver tissue of CE patients, compared to normal liver. Incubation of LX-2 cells (in vitro) with HCF induced a decreased proliferation of these cells and a reduced expression of miR-19, inversely correlated with the expression of collagen 1A1 and TGF-ß receptor II (TßRII). Conversely, overexpression of miR-19 by LX-2 cells inhibited the proliferation of these cells and led to decreased TßRII expression. CONCLUSIONS: Our study provides new evidence for the intervention of miRNAs in the regulation of fibrosis in infectious diseases; it suggests that E. granulosus can inhibit miR-19 liver expression and promote fibrosis through the increase in TßRII, the activation of hepatic stellate cells and extracellular matrix production.

Ultrastructural characterization of craniopharyngioma at the tumor boundary: A structural comparison with an experimental toxic model using "oil machinery" fluid, with emphasis on Rosenthal fibers.

Craniopharyngiomas (CPs) are cystic, encapsulated, slow-growing epithelial tumors. CPs can be aggressive forms invading and resorting surrounding structures of adjacent brain tissue, where Rosenthal fibers (RFs) are expressed. The aim of this study was to investigate the ultrastructure of these fibers in human biopsies and compare it with an experimental toxic model produced by the cortical infusion of the oil cyst fluid ("Oil machinery" fluid or OMF) from CPs to rats. For this purpose, the CPs from ten patients were examined by light and electron microscopy. OMF was administered to rats intracortically. Immunohistochemical detection of glial fibrillary acidic protein (GFAP) and vimentin was assessed. In both freshly obtained CPs and rat brain tissue, the presence of abundant cellular debris, lipid-laden macrophages, reactive gliosis, inflammation and extracellular matrix destruction were seen. Ultrastructural results suggest focal pathological disturbances and an altered microenvironment surrounding the tumor-brain junction, with an enhanced presence of RFs in human tumors. In contrast, in the rat brain different degrees of cellular disorganization with aberrant filament-filament interactions and protein aggregation were seen, although RFs were absent. Our immunohistochemical findings in CPs also revealed an enhanced expression of GFAP and vimentin in RFs at the peripheral, but not at the central (body) level. Through these findings we hypothesize that the continuous OMF release at the CPs boundary may cause tissue alterations, including damaging of the extracellular matrix, and possibly contributing to RFs formation, a condition that was not possible to reproduce in the experimental model. The presence of RFs at the CPs boundary might be considered as a major criterion for the degree of CPs invasiveness to normal tissue. The lack of RFs reactivity in the experimental model reveals that the invasive component of CPs is not present in the OMF, although the fluid per se can exert tissue damage.

Increased expression of secreted frizzled-related protein 4 in polycystic kidneys.

Autosomal dominant polycystic kidney disease (ADPKD) is a common hereditary disease associated with progressive renal failure. Although cyst growth and compression of surrounding tissue may account for some loss of renal tissue, the other factors contributing to the progressive renal failure in patients with ADPKD are incompletely understood. Here, we report that secreted frizzled-related protein 4 (sFRP4) is upregulated in human ADPKD and in four different animal models of PKD, suggesting that sFRP4 expression is triggered by a common mechanism that underlies cyst formation. Cyst fluid from ADPKD kidneys activated the sFRP4 promoter and induced production of sFRP4 protein in renal tubular epithelial cell lines. Antagonism of the vasopressin 2 receptor blocked both promoter activity and tubular sFRP4 expression. In addition, sFRP4 selectively influenced members of the canonical Wnt signaling cascade and promoted cystogenesis of the zebrafish pronephros. sFRP4 was detected in the urine of both patients and animals with PKD, suggesting that sFRP4 may be a potential biomarker for monitoring the progression of ADPKD. Taken together, these observations suggest a potential role for SFRP4 in the pathogenesis of ADPKD.

Relation between size of odontogenic jaw cysts and the pressure of fluid within.

Intracystic fluid pressure may have a crucial role in the growth of odontogenic jaw cysts. In this study, we investigated the relation between the size of the cyst and the pressure of the fluid within odontogenic keratocysts, dentigerous cysts, and radicular cysts. The radiolucent area of the cyst on a panoramic radiograph was linearly related to the volume in the cavity, and the correlation coefficient (gamma) was 0.70 (n = 25, P < 0.001). Intracystic fluid pressure correlated negatively with the radiolucent area in odontogenic keratocysts (gamma = -0.76, n = 9, P = 0.02), dentigerous cysts (gamma = -0.54, n = 16, P = 0.03), and radicular cysts (gamma = -0.69, n = 10, P = 0.03). The values of [(intracystic fluid pressure (mmHg)) x (radiolucent area (cm(2)))] did not differ significantly among the three types of cyst. Intracystic fluid pressure may therefore be negatively related to the size of all three types of cyst.

Hepatocyte growth factor (HGF) in ovarian epithelial tumour fluids stimulates the migration of ovarian carcinoma cells.

Hepatocyte growth factor (HGF) is a pleiotropic growth factor implicated in the growth and spread of some epithelial tumours. The epithelial cells of a proportion of ovarian tumours, and some ovarian carcinoma cell lines, express high levels of the HGF receptor, c-Met. In this study, we show that ovarian ascitic fluid as well as benign and malignant ovarian cyst fluids contain significant levels of HGF. Ovarian cyst and ascitic fluid stimulated the migration of the ovarian carcinoma cell line, SK-OV-3, and this was greatly reduced by the addition of an HGF-neutralising antibody. Non-malignant peritoneal fluid contained low levels of HGF, and did not stimulate migration of the SK-OV-3 cells. Our results show that HGF is present in benign and malignant ovarian cyst and ascitic fluid, and that HGF in ovarian tumour fluid may be a major inducer of ovarian carcinoma cell migration.

Effects of chocolate cyst fluid on endometrioma cell growth in culture.

OBJECTIVE: To evaluate the effect of chocolate cyst fluid on the proliferation of cultured human endometrioma cells and to assay the concentration of transforming growth factor-B1 in this fluid. DESIGN: Controlled in vitro study. SETTING: Department of Obstetrics and Gynecology, State University of New York Health Science Center. PATIENT(S): Five women with ovarian endometriomas. INTERVENTION(S): Endometrioma tissue and chocolate fluid from five different patients were entered in this study. MAIN OUTCOME MEASURE(S): Endometrioma cell proliferation in culture with and without chocolate cyst fluid. RESULT(S): Chocolate cyst fluid increased the proliferation of endometrioma cells compared with controls. Also, high concentrations of transforming growth factor-B1 were found in cysts' fluid. CONCLUSIONS: Chocolate cyst fluid has a growth-enhancing effect on endometrioma cells. One promoting growth factor is transforming growth factor-B1.