Fibrotic Stage Research Articles

Role of microRNA-210-3p in hepatitis B virus-related hepatocellular carcinoma.

Hepatitis B virus (HBV)-related hepatocarcinogenesis is not necessarily associated with the liver fibrotic stage and is occasionally seen at early fibrotic stages. MicroRNAs (miRNAs) are essentially 18- to 22-nucleotide-long endogenous noncoding RNAs. Aberrant miRNA expression is a common feature of various human cancers. The aberrant expression of specific miRNAs has been shown in hepatocellular carcinoma (HCC) tissue compared with nontumor tissue. Thus, we examined targetable miRNAs as a potential new biomarker related to the high risk of HBV-related hepatocarcinogenesis, toward the prevention of cancer-related deaths. HCC tissue samples from 29 patients who underwent hepatectomy at our hospital in 2002-2013 were obtained. We extracted the total RNA and analyzed it by microRNA array, real-time RT-PCR, and three comparisons: 1) HBV-related HCC and adjacent nontumor tissue, 2) HCV-related HCC and adjacent nontumor tissue, and 3) non-HBV-, non-HCV-related HCC and adjacent nontumor tissue. We also performed a functional analysis of miRNAs specific for HBV-related HCC by using HBV-positive HCC cell lines. MiR-210-3p expression was significantly increased only in the HBV-related HCC tissue samples. MiR-210-3p expression was upregulated, and the levels of its target genes were reduced in the HBV-positive HCC cells. The inhibition of miR-210-3p enhanced its target gene expression in the HBV-positive HCC cells. In addition, miR-210-3p regulated the HBx expression in HBV-infected Huh7/NTCP cells. The enhanced expression of miR-210-3p was detected specifically in HBV-related HCC and regulated various target genes, including HBx in the HBV-positive HCC cells. MiR-210-3p might, thus, be a new biomarker for the risk of HBV-related HCC.NEW & NOTEWORTHY Our present study demonstrated that miR-210-3p is the only microRNA with enhanced expression in HBV-related HCC, and the enhanced expression of miR-210-3p upregulates HBx expression. Therefore, miR-210-3p might be a pivotal biomarker of HBV-related hepatocarcinogenesis, and the inhibition of miR-210-3p could prevent inducing hepatocarcinogenesis related to HBV infection.

P703 A breathtaking heart. The loeffler"s endocarditis

Abstract This is a 72 year-old man known for idiopathic HES with endomyocardial fibrosis and moderate aortic regurgitation, treated with corticosteroids and antithrombotics. Two months prior to admission, he develops progressive exertional dyspnea reaching NYHA 3 class, with concomitant peripheral edema and rise in NT-proBNP. Cardiac transthoracic work-up shows a preserved left ventricular ejection fraction, but more marked dilatation with decrease in left ventricular volume due to a large echodense mass in the apex. The transmitral flow is restrictive, and the aortic regurgitation moderate. In view of this rapid evolution, resection of the left ventricular mass with concomitant aortic valve replacement is performed. Pathology confirmed eosinophilic infiltration. First described in 1975, hypereosinophilic syndrome (HES) is a rare pathology that is defined as an absolute eosinophil count (>1,5 G/L) in the peripheral blood with eosinophil-mediated organ damage and /or dysfunction. We know three mains categories; primary (or neoplastic), secondary (or reactive) and idiopathic. Cardiac involvement is uncertain and cardiac injury does not clearly correlate with degree of peripheral eosinophilia. Heart tissue damage evolves through three stages (necrotic, intermediate and fibrotic). Endomyocardial fibrosis and eosinophilic myocarditis, also known as "Loeffler’s endocarditis", are major causes of morbidity and mortality among patients with HES. Echocardiography plays a crucial role in initial diagnosis of endomyocardial fibrosis, displaying myocardial wall thickening, signs of restriction (mitral inflow with large E wave and small A wave; E/A ratio greater than 2, short deceleration time less than 150 ms) and intracardiac thrombus (fixed mass embedding left ventricular apex). Valve regurgitation is another possible complication due to entrapment of the chordae tendinae and/or leaflets during the fibrotic stage of this disease. Cardiac MRI is another important diagnostic tool for precising cardiac involvement. Despite of steadily improving noninvasive diagnostic imaging methods, endomyocardial biosy still remains the gold standard exam and ultrasounds work-up is crucial for follow-up. In conclusion, endomyocardial fibrosis is a rare cause of restrictive cardiomyopathy characterized by echocardiographic sign of elevated filling pressure, progressive endomyocardial thickening, valve regurgitation and possible intracardiac thrombus formation. This case underlines the importance of echocardiography in initial diagnosis and regular follow-up of this type of patients, in order to adapt medical treatment and monitor hemodynamic evolution of the restrictive physiology and of valvular damage. Abstract P703 Figure. HES-3

Dissecting the Functional Reprogramming of the Microenvironment in Bone Marrow Fibrosis at the Single Cell Level

Functional contributions of individual members of the bone marrow microenvironment to myelofibrosis (MF) in patients with myeloproliferative neoplasms (MPN) are incompletely understood. Consequently, we aimed to generate a comprehensive map of the stroma in MPN/MF on a single cell level. Our analysis revealed two distinct mesenchymal stromal cells (MSC) subsets as pro-fibrotic cells. MSCs were functionally reprogrammed in a stage-dependent manner with loss of their progenitor status and initiation of differentiation in the pre-fibrotic stage and acquisition of a pro-fibrotic and inflammatory phenotype in the fibrotic stage. In parallel, IL-33-expressing myelinating Schwann cell progenitors expanded, likely as a repair mechanism for the previously described neuropathy in MPN. IL-33 had anti-fibrotic effects on MSCs but induced expression of the alarmin S100A8 leading to loss-of-hematopoiesis support. Importantly, we demonstrate that S100A8 expression in stroma and patient plasma marks fibrosis progression and might even serve as a biomarker for disease progression.

Survival of endogenous hepatic stem/progenitor cells in liver tissues during liver cirrhosis

AimsThis study addressed whether endogenous hepatic stem/progenitor (HSP) cells survival were related to the injured signals during liver cirrhosis. Material and methodsLiver cirrhosis was induced in C57BL/6 mice by administering diethylnitrosamine (DEN) in drinking water. Hematoxylin-eosin staining and Masson's trichrome staining were used to identify infiltrative cells and connective tissues, respectively. The inflammatory activity grade and fibrotic stage, represented as G and S respectively, and evaluated by the International Simplified Grading and Staging System (ISGSS). Endogenous HSP cells (Ng2+HSP) were identified by immunofluorescence staining with an anti-neuro/glial antigen 2 (Ng2) antibody. All data were analyzed using SPSS 22.0 and GraphPad Prism 6 and Student's t-test was performed to determine statistical significance. p-values < 0.05 were considered statistically significant. Key findingsAll mice administered oral DEN developed liver fibrosis, liver cirrhosis and hepatocellular carcinoma (HCC). During the fibrosis period, observed a positive correlation of survival of endogenous HSP (Ng2+HSP) cells with inflammatory activity. As the disease developed into the cirrhotic stage, when lost correlation of endogenous HSP cells with inflammatory activity, revealed a dramatically reduced survival rate of endogenous HSP (Ng2+HSP) cells. SignificanceThe DEN-induced liver cirrhosis could develop into three time zone of fibrosis, cirrhosis and cancer, and the histological patterns in the model are similar to those described in humans. Dramatic survival and less apoptosis of endogenous HSP (Ng2+HSP) cells was within fibrotic state, where inflammation signals is strong, indicating such time zone (1–6 weeks) during liver cirrhosis is important for mobilizing endogenous HSP-based regeneration.

Changes in the pattern of rhinoscleroma in Egypt: updated experience.

This study aimed to record recent changes in the incidence and clinical and histopathological patterns of rhinoscleroma in the middle of Nile Delta in Egypt. Twenty-eight patients with rhinoscleroma were included in this study. Patients who were attending the Department of Otolaryngology at Tanta University Hospitals from January 2014 to January 2019 were included. Cases were diagnosed by clinical, bacteriological and histopathological data. Nasal biopsies were taken for histopathological and electron microscopy studies. All patients received a twelve-week course of 500 mg ciprofloxacin tablets twice daily. The rate of rhinoscleroma was shown to be decreasing compared with previous studies. In this study, the disease mainly affected the nose without involvement of the palate, lips, Eustachian tube or bronchi. Histopathological and electron microscopy analyses showed that most cases were in the atrophic and fibrotic stages rather than the granulomatous stage of disease. The incidence of rhinoscleroma in Egypt is decreasing and its clinical and histopathological patterns are changing to less advanced forms. This may be explained by the improvement in socioeconomic conditions and methods of treatment.

Exploring the Diagnostic Potential of Serum Golgi Protein 73 for Hepatic Necroinflammation and Fibrosis in Chronic HCV Infection with Different Stages of Liver Injuries.

Background and Aim Serum Golgi protein 73 (GP73) is a promising alternative biomarker of chronic liver diseases, but most data are from patients with HBV infection rather than HCV. Materials and Methods Two independent cohorts of chronic hepatitis C (CHC) patients from the 5th Medical Centre of the Chinese PLA General Hospital (n = 174) and Beijing Youan Hospital (n = 120) with different histories of HCV infection were enrolled. The correlations between serum GP73 and other biochemical indices, as well as its correlations with different stages of liver disease progression, were investigated. The receiver operating characteristic (ROC) curve was employed to evaluate the diagnostic potential of serum GP73 for liver necroinflammation and fibrosis, and comparisons of the diagnostic efficiency with traditional indices of hepatic liver injuries were further investigated. Results Levels of serum GP73 were found significantly elevated in patients with moderate to severe inflammatory grade (G ≥ 2) and/or with advanced fibrotic stages (F ≥ 3) in both cohorts (P < 0.05, respectively), as compared to those with a normal or mild liver lesion. Further ROC analysis demonstrated that serum GP73 was comparable to serum ALT and AST in diagnosing the liver necroinflammation grade at G ≥ 2, but its diagnostic values for advanced fibrosis (F ≥ 3) and cirrhosis (F = 4) were limited when compared to APRI and FIB-4, and FIB-4 exhibited the best performance. Notably, an obvious elevation of serum GP73 was observed after patients received PEG-IFN and ribavirin treatment. Conclusions Serum GP73 is an important biomarker in evaluating and monitoring the disease progression including liver necroinflammation and fibrosis in patients with chronic HCV infection, but the value is limited for diagnosing advanced fibrosis and cirrhosis in comparison with APRI and FIB-4.

IL-33 promotes the progression of nonrheumatic aortic valve stenosis via inducing differential phenotypic transition in valvular interstitial cells

ObjectiveInterleukin (IL)-33 is a mediator in the pathogenesis of several inflammatory diseases. Its receptor, ST2, is overexpressed in nonrheumatic aortic valve stenosis (NR-AS). This study compared smooth muscle α-actin (α-SMA), osteopontin (OPN), and suppression of tumorigenicity 2 (ST2) expression between specimens from fibrotic and calcific stages of NR-AS and observed the effects and mechanisms of phenotypic transition of porcine valvular interstitial cells (VICs) in the presence of IL-33. MethodsPeripheral blood IL-1 family mRNA and protein levels in NR-AS patients and healthy adults were quantified by real-time quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay. Immunohistochemistry and immunofluorescence were used to detect the expression and coexpression of α-SMA, OPN, and ST2 in NR-AS specimens. Porcine VICs were stimulated with IL-33, IL-33+SB203580, or IL-33+SC75741. mRNA and protein expression levels of porcine VICs were detected by RT-qPCR and western blot. ResultsThe mRNA and protein levels of IL-33 and sST2 in peripheral blood of NR-AS patients were higher than those in healthy adults. Immunohistochemistry and immunofluorescence showed higher expression of α-SMA, OPN, and ST2 in the calcific stage of NR-AS than in the fibrotic stage. Coexpression of ST2/α-SMA or ST2/OPN was found only in the calcific stage. Nuclear factor (NF)-κB and p38 mitogen-activated protein kinase (MAPK) phosphorylation levels were associated with IL-33-induced porcine VIC differentiation into myofibroblasts and osteoblasts, respectively. IL-33 stimulation also promoted the coexpression of ST2/OPN or α-SMA/OPN/ST2. ConclusionIL-33 might be a potential biomarker for NR-AS. IL-33-induced porcine VIC differential phenotypic transition and differentiation into myofibroblasts and osteoblasts were dependent on the NF-κB and p38 MAPK signaling pathways, respectively.

Evaluation of aspartate aminotransferase to platelet ratio index and fibrosis 4 scores for hepatic fibrosis assessment compared with transient elastography in chronic hepatitis C patients.

Background and AimFibrotic stage (FS) assessment is essential in chronic hepatitis C treatment cascade. Liver stiffness measurement (LSM) using transient elastography (TE) is reliable and correlated with liver biopsy. However, TE may not be widely available. This study aimed to evaluate the diagnostic performances of aspartate aminotransferase to platelet ratio index (APRI) and fibrosis 4 (FIB‐4) scores compared with TE.MethodsWe conducted a multicenter, cross‐sectional study, including all chronic hepatitis C virus (HCV) monoinfection patients with successful and reliable LSM, at 10 centers in Thailand from 2012 to 2017. Characteristics and laboratory data within 3 months of TE were retrospectively reviewed. Using TE as a reference standard, the diagnostic performances of APRI and FIB‐4 were evaluated. TE cut‐off levels of 7.1 and 12.5 kPa represented significant fibrosis (SF) and cirrhosis, respectively.ResultsThe distribution of FS by TE in 2000 eligible patients was as follows: no SF 28.3%, SF 31.4%, and cirrhosis 40.3%. APRI ≥ 1 provided 70.1% sensitivity and 80.6% specificity, with an area under the receiver operator characteristics curve (AUROC) of 0.834 for cirrhosis. The specificity increased to 96.3% when using a cut‐off level of APRI ≥ 2. FIB‐4 ≥ 1.45 provided a sensitivity, specificity, and AUROC of 52.4%, 91.0%, and 0.829 for cirrhosis, respectively. For SF, APRI performed better than FIB‐4, with an AUROC of 0.84 versus 0.80 (P < 0.001). APRI score < 0.5 and FIB‐4 score > 1.45 yielded sensitivities of 82.3% and 74.4% and specificities of 65.4% and 69.8%, respectively.ConclusionsAPRI and FIB‐4 scores had good diagnostic performances for FS assessment compared with TE, especially for cirrhosis. APRI may be used as the noninvasive assessment in resource‐limited settings for HCV patients’ management.

Immunopathogenesis of Juvenile Systemic Sclerosis.

Juvenile-onset systemic sclerosis (jSSc) is a rare and severe autoimmune disease with associated life-threatening organ inflammation and evidence of fibrosis. The organ manifestations of jSSc resemble adult SSc, but with better outcomes and survival. The etiology of jSSc appears to reflect adult-onset SSc, with similar inflammatory mediators and autoantibodies, but with a significant population of children with uncharacterized anti-nuclear antibodies. The genetics of patients with jSSc differ from women with SSc, resembling instead the genes of adult males with SSc, with additional HLA genes uniquely associated with childhood-onset disease. Current treatments are aimed at inhibiting the inflammatory aspect of disease, but important mechanisms of fibrosis regulated by dermal white adipose tissue dendritic cells may provide an avenue for targeting and potentially reversing the fibrotic stage.

PF667 THE ROLE OF HEPCIDIN AND DYSREGULATED IRON HOMEOSTASIS IN THE PATHOGENESIS OF ANEMIA IN MYELOFIBROSIS

Background:Myelofibrosis (MF) is a stem cell disorder with impaired hematopoiesis and complex pathogenesis. Anemia in MF is a result of a multifactorial process, which is incompletely understood. The central pathogenetic mechanism is replacement of normal hemopoietic tissue by fibrotic stroma. However, the ineffective erythropoiesis, inflammation and iron overload have an additional impact on development and worsening of the anemia, suggesting the role of dysregulated iron homeostasis and hepcidin.Aims:to analyze hepcidin, IL‐6, IL‐8 levels and parameters of iron metabolism in a random cross‐section of patients with different forms and stages of myelofibrosis.Methods:We analyzed 36 patients with MF – 75 % with primary MF(PMF) (prefibrotic PMF – 38,9% and overt PMF – 36,1%) and 25% with secondary MF (post‐PV −11,1% and post‐ET – 13,9%); median age of the group was 68,5 years (range 39–88); male: female ratio‐ 1,57:1. According to DIPSS: 5,6% were Low; 52,8% were Intermediate 1; 33,3% were Intermediate 2 and 8,3% were High risk. Regarding treatment 30,6% of the patients received hydroxyurea, 8,3% ‐ Interferon, 19,4% ‐ Ruxolitinib and 41,7% ‐ best supportive care. Patients who were transfused with ≥3 RBC units/month were 19,5% and those receiving &lt;3 RBC units/month ‐ 80,5%. All patients were divided in 3 groups according to the time from diagnosis: newly diagnosed (30,6%), 1 to 5 years (47,2%) and &gt; 5 years (22,2%) from diagnosis. Serum ferritin, Fe, TIBC and parameters of CBC were measured as a part of routine clinical assessment. Serum hepcidin and serum concentrations of IL 6 and IL8 were measured in duplicate by ELISA (My BioSource, San Diego, USA) in all patients and 14 healthy controls.Results:In our study the levels of hepcidin in MF patients are not found statistically different from levels in healthy controls. No difference is found in hepcidin level within the three risk groups according to DIPSS (Intermediate 1, Intermediate 2 and High), neither between the subtypes of disease (proliferative or fibrotic stage of primary myelofibrosis and secondary myelofibrosis). However, we find significant difference between hepcidin levels within the three groups according to disease duration. Patients with newly diagnosed MF have higher levels of hepcidin compared to those with prolonged evolution (F = 7,09, R = −0,476, R2 = 0,226, p = 0,003). Patients receiving &gt; 3 RBC transfusions/month present with lower hepcidin compared to transfusion independent patients (10,10 ± 6,67; 31,15 ± 44,37; p &lt; 0,05). We also find significantly lower hepcidin level in patients receiving cytoreductive or target treatment compared to patients without any therapy (17,74 ± 21,99; 43,05 ± 58,46; p &lt; 0,05). In all risk groups hepcidin positively correlates with leukocytosis and age. Unexpectedly, higher levels of ferritin and serum iron are associated with low hepcidin. In multivariate analysis a significant straight correlation between hepcidin level and IL6 (R = 0,427, p = 0,009) and between IL6 and IL8 levels in the whole cohort (R = 0,500, p = 0,002) is found.Summary/Conclusion:The hepcidin regulation is complex and multifactorial. Its role in pathogenesis of anemia in myelofibrosis is controversary. Probably it has a higher impact in early stages of the disease and depends on treatment and transfusions. A further analysis, including labile iron, would better clarify the precise mechanisms of anemia in a heterogeneous disorder like MF.

FRI-302-Impact of exercise on advanced fibrotic stage of NASH and liver carcinogenesis

FRI-302-Impact of exercise on advanced fibrotic stage of NASH and liver carcinogenesis

Serum Proteomic Changes as Candidate Biomarkers of Intermediate Liver Fibrosis in Chronic Hepatitis B Infection.

Chronic hepatitis B (CHB) is a major global health burden. Liver fibrosis, an insidious process, is the main histopathological change in CHB that might lead to the end-stage liver disease if left untreated. The intermediate liver fibrosis (S2) is the optimal time to start antiviral therapy. The aim of the present study was to examine the proteomic changes in patients with CHB at different fibrotic stages, with a view to identify future serum biomarkers for S2. Ninety CHB patients were grouped into mild (S0-1), intermediate (S2), and severe liver fibrosis (S3-4) (61 men and 29 women; age 25-63 years). Isobaric tagging for relative and absolute quantitation was applied to screen proteins differentially expressed among the patient groups. Another 46 patients with CHB (age 25-59 years; 31 men and 15 women), and 16 healthy controls (age 26-61 years; 11 men and 5 women) were enrolled in a validation group. Enzyme-linked immunosorbent assay was used to verify the diagnostic value of the candidate biomarkers. We found 139 proteins that were differentially expressed between various fibrotic stage-paired comparisons. Five protein candidates were selected as potential biomarkers of S2 for further verification. Notably, ficolin-2 (FCN2) and carboxypeptidase B2 (CPB2) showed differential expression between patients and healthy controls. In conclusion, serum proteomic changes reported here offer new molecular leads for future research on biomarker candidates to identify liver fibrotic stages in CHB. In particular, FCN2 and CPB2 warrant further research on their possible mechanistic involvement in CHB pathogenesis.

Delayed resolution of bleomycin-induced pulmonary fibrosis in absence of MMP13 (collagenase 3)

Matrix metalloprotease 13 (MMP13) deficiency in pulmonary fibrosis has described contradictory phenotypes on inflammatory and fibrotic responses after lung injury, and its role during lung fibrosis resolution is still undefined. MMP13 has been considered the main collagenase in rodents, and the remodeling of fibrillar collagen is widely attributed to the action of this enzyme. In this study we aimed to explore the role of MMP13 during lung fibrosis progression and resolution. Lung fibrosis was induced by intratracheal instillation, and inflammatory, fibrotic, and resolution stages were evaluated in Mmp13-null and wild-type (WT) mice. Bronchoalveolar lavage fluid was taken for cytokine array analysis and activity of gelatinases. Our results showed that MMP13 is upregulated mainly during two stages after lung injury, inflammation and resolution of fibrosis, and it is mainly expressed by alveolar and interstitial macrophages. Mmp13-null mice exhibited more extensive inflammation at 7 days after bleomycin treatment, and it was characterized by increased macrophage infiltration and significant alterations in proinflammatory cytokines. We also documented that Mmp13-deficient mice experienced more severe and prolonged lung fibrosis compared with WT mice. Delayed resolution in Mmp13-deficient lungs was characterized by a decreased overall collagenolytic activity and persistent fibrotic foci associated with emphysema-like areas. Together, our findings indicate that MMP13 plays an antifibrotic role and its activity is crucial in lung repair and restoration of tissue integrity during fibrosis resolution.

Adipokine expression in systemic sclerosis lung and gastrointestinal organ involvement

ObjectivesThe immunomodulatory properties of adipokines have previously been reported in autoimmune disorders. Less is known about the role of adipokines in systemic sclerosis (SSc). Lung and gastrointestinal tract are frequently involved in SSc; therefore, these organs were analyzed for adipokine expression as well as pulmonary samples of patients suffering from idiopathic pulmonary fibrosis (IPF) as comparison. MethodsGastric samples (antrum, corpus) of SSc were analyzed immunohistochemically for adiponectin, resistin and visfatin compared with non-SSc related gastritis. Inflammatory cells were quantified in gastric samples and correlated with adipokine expression. Lung samples of SSc, IPF and healthy controls were also analyzed. Protein levels of lung tissue lysates and bronchoalveolar lavages (BAL) in minor fibrotic stages were measured by ELISA. ResultsLung sections of donor parenchyma showed significantly stronger adiponectin signals as IPF and SSc (donor vs. IPF: p < 0.0001). In SSc and IPF, resistin and visfatin were increased within immune cell infiltrates, but overall no difference in expression for resistin or visfatin compared to controls was observed. In BAL and lung protein lysates of early stages of fibrosis, adiponectin and visfatin were not reduced in IPF and SSc compared to controls. In gastric samples collected by standard endoscopic gastric biopsy, adiponectin was also significantly reduced in SSc- compared to non-SSc gastritis (p = 0.049) while resistin and visfatin were comparable although deeper fibrotic layers were not included in the respective samples. Adiponectin-positive tissues showed higher amounts of CD4+ but not CD8+ T cells. Controls showed no correlation between CD4+ T cells and resistin, whereas SSc showed significantly more CD4+ T cells in resistin-negative tissues. ConclusionAdipokines are expressed in gastric and lung samples of patients with SSc and in lung samples affected by IPF. Prominently, adiponectin levels were reduced in fibrotic SSc gastritic tissue as well as in IPF and SSc lung tissue. Consequently, adiponectin expression seems to be associated with fibrotic progression in the context of SSc and IPF.

Visualisation of interstitial lung disease by molecular imaging of integrin αvβ3 and somatostatin receptor 2

ObjectiveTo evaluate integrin αvβ3 (alpha-v-beta-3)-targeted and somatostatin receptor 2 (SSTR2)-targeted nuclear imaging for the visualisation of interstitial lung disease (ILD).MethodsThe pulmonary expression of integrin αvβ3 and SSTR2 was analysed in...

Clinical Analysis of Statistical Data on the Characteristics of Patients with Acute Respiratory Distress Syndrome in a Greek ICU.

Acute respiratory distress syndrome (ARDS) is a potentially life-threatening condition for a many patients. The associated mortality rate may exceed 50% in some cases. The etiopathogenesis of the syndrome remains unclear, and in Greece the exact number of people suffering or dying of ARDS each year is unknown. In this study, we sought to record the clinical characteristics and outcomes of all patients with laboratory confirmed ARDS. This is a retrospective single-center observational study, conducted in mixed, closed-type ICU with 12 beds at a Greek public health system hospital. The medical records of patients admitted to the ICU from January 1, 2015, until December 31, 2016, were retrospectively studied, and statistical analyses were performed using SPSS version 22.0 software. Patients were assessed using the Fisher exact test assuming variances. All tests were two-tailed. In total, 15 patients developed ARDS (5.4%). According to the existence of radiographic pulmonary edema image with bilateral infiltrates, 12 patients were classified in the early phase of the syndrome described by the term exudative phase and 3 patients were classified in the third stage of final fibrosis (fibrotic stage). According to the Fisher test, there is a significant correlation between age and outcome. The incidence of the syndrome among hospitalized patients in ICU is in line with the current literature, while the main predisposing factor in this study was acute pancreatitis.

Impaired LXRα Phosphorylation Attenuates Progression of Fatty Liver Disease

SummaryNon-alcoholic fatty liver disease (NAFLD) is a very common indication for liver transplantation. How fat-rich diets promote progression from fatty liver to more damaging inflammatory and fibrotic stages is poorly understood. Here, we show that disrupting phosphorylation at Ser196 (S196A) in the liver X receptor alpha (LXRα, NR1H3) retards NAFLD progression in mice on a high-fat-high-cholesterol diet. Mechanistically, this is explained by key histone acetylation (H3K27) and transcriptional changes in pro-fibrotic and pro-inflammatory genes. Furthermore, S196A-LXRα expression reveals the regulation of novel diet-specific LXRα-responsive genes, including the induction of Ces1f, implicated in the breakdown of hepatic lipids. This involves induced H3K27 acetylation and altered LXR and TBLR1 cofactor occupancy at the Ces1f gene in S196A fatty livers. Overall, impaired Ser196-LXRα phosphorylation acts as a novel nutritional molecular sensor that profoundly alters the hepatic H3K27 acetylome and transcriptome during NAFLD progression placing LXRα phosphorylation as an alternative anti-inflammatory or anti-fibrotic therapeutic target.

Surgical outcomes of immediate sequential bilateral vitreoretinal surgery for advancing retinopathy of prematurity.

Purpose:Bilateral eye surgery in the same session may be required for advancing stage 4 retinopathy of prematurity (ROP). The purpose of this study was to evaluate the outcomes of immediate sequential bilateral vitreoretinal surgery (ISBVS) in stage 4 ROP.Methods:In a retrospective interventional study at a tertiary care center, 60 eyes of 30 infants who underwent ISBVS for stage 4 ROP between December 2015 and May 2017 were studied. In cases with clear retrolental access, 25G or 27G lens sparing vitrectomy (LSV) was performed and in the rest 25G lensectomy with vitrectomy (LV) was performed through clear corneal entries. The final anatomical outcome measures were the status of tractional retinal detachment (TRD) and macular status.Results:The mean gestational age was 28.4 ± 2.0 weeks and birth weight was 1214.5 ± 329.7gms. The mean postconceptional age at surgery was 40.8 ± 2.2 weeks. Stages 4a and 4b were present in 86.7% and 13.3% eyes respectively. LSV was performed in 95% eyes whereas LV was performed in the rest. None of the eyes developed lens touch, choroidal hemorrhage, postoperative hypotony, corneal decompensation, or endophthalmitis. At last follow-up (mean 45 weeks, range 20–68 weeks), macula was attached in 90% eyes with the TRD resolved completely in 61.7% eyes and significantly decreased in another 25% eyes. Sequalae included macular drag, epiretinal membrane, and progression to fibrotic stage 5 disease.Conclusion:ISBVS is safe and effective for bilateral stage 4 ROP and should be recommended in rapidly progressive cases.

Screening of antifibrotic compounds using myofibroblasts

Idiopathic pulmonary fibrosis (IPF) is a chronic and progressive disease of unknown cause. Under the pathogenic environment, myofibroblasts (MyoFs) mainly differentiated from fibroblasts play a key role in lung fibrogenesis. Established MyoF has been considered as an irreversible phenotype, but recently shown to dedifferentiate to fibroblast. This feature is desirable in development of pharmacologic strategy against IPF because most patients with IPF appear to accumulate pathogenic MyoFs in their lungs at the time of clinical presentation. Therefore, we have established several strains of primary cultured MyoFs from the fibrotic lungs of patients. Using our MyoF assay system, the drug library for compounds that inhibit epigenetics-related signals was screened by monitoring downregulation in expression of collagen and MyoF markers, α-SMA and ED-A fibronectin. Through this assay, we found in vitro that a certain histone methyltransferase inhibitor potently dedifferentiated MyoFs. In addition, intratracheal administration with the compound at the early fibrotic stage of bleomycin-injured lung successfully ameliorated lung fibrosis in mice. We will discuss the mechanism by which the compound affects pathogenic myofibroblast under lung fibrogenesis.

Clinicopathological and molecular features of SF3B1-mutated myeloproliferative neoplasms

The introduction of next-generation sequencing has broadened the genetic landscape of myeloproliferative neoplasms (MPNs) beyond JAK2, MPL, and CALR. However, the biological role and clinical impact of most other mutations are not well defined. We interrogated 101 genes in 143 BCR-ABL1-negative MPNs in chronic phase from 2 large institutions. We detected SF3B1 mutations in 15 cases (10%) and set to investigate the clinical, morphologic, and molecular features of SF3B1 mutated (SF3B1+) MPNs in comparison to SF3B1 wild-type (SF3B1-) cases and to identify distinctive features with myelodysplastic/myeloproliferative neoplasms with ring sideroblasts (RS) and thrombocytosis, which can show partial clinical and morphological overlap with MPNs. SF3B1+ cases were enriched in primary myelofibrosis in both prefibrotic and fibrotic stage, but mutations of SF3B1 seem to occur only as a late event in the fibrotic phase of essential thrombocythemia and polycythemia vera. SF3B1+ MPNs showed borderline lower hemoglobin but no other clinical or molecular differences compared to SF3B1- MPNs. Of note, RS were present only in a subset of SF3B1+ cases (4/10) without any other feature of erythroid or granulocytic dysplasia. Our results suggest that mutations in SF3B1 are not a rare event in MPNs, especially in primary myelofibrosis and during late fibrotic stages of essential thrombocythemia and polycythemia vera, but are not associated with myelodysplastic progression. Careful examination of bone marrow and peripheral blood for morphologic dysplasia is crucial to reach the correct diagnosis and avoid a misdiagnosis of myelodysplastic/myeloproliferative neoplasms with RS and thrombocytosis, a pitfall with potential prognostic and therapeutic implications.