RelB activation in anti-inflammatory decidual endothelial cells: a master plan to avoid pregnancy failure?

It is known that excessive inflammation at fetal-maternal interface is a key contributor in a compromised pregnancy. Female genital tract is constantly in contact with microorganisms and several strategies must be adopted to avoid pregnancy failure. Decidual endothelial cells (DECs) lining decidual microvascular vessels are the first cells that interact with pro-inflammatory stimuli released into the environment by microorganisms derived from gestational tissues or systemic circulation. Here, we show that DECs are hypo-responsive to LPS stimulation in terms of IL-6, CXCL8 and CCL2 production. Our results demonstrate that DECs express low levels of TLR4 and are characterized by a strong constitutive activation of the non-canonical NF-\u3baB pathway and a low responsiveness of the canonical pathway to LPS. In conclusion, DECs show a unique hypo-responsive phenotype to the pro-inflammatory stimulus LPS in order to control the inflammatory response at feto-maternal interface

Activation of hepatic stem cells compartment during hepatocarcinogenesis in a HBsAg HBV-transgenic mouse model

Abstract Chronic infection of Hepatitis B Virus (HBV) is one of the highest risk factors of hepatocellular carcinoma (HCC). The accumulation of HBV surface antigen (HBsAg) into hepatocytes induces inflammation and oxidative stress, impairing their replicative ability and allowing the activation of the hepatic stem cells (SC) compartment. This study aimed to understand the involvement of SC during hepatocarcinogenesis in HBsAg-related liver damage, from early injury until HCC. HBsAg-transgenic (TG) and wild type (WT) mouse were followed at several stages of the liver damage: inflammation, early hepatocytes damage, dysplasia, and HCC. Serum transaminases, liver histology, and diagnostic data were collected. The expressions of SC and cancer stem cells (CSC) markers was analyzed by RT-qPCR, immunohistochemistry and Western blot. Starting from 3 months, TG animals showed a progressive liver damage characterized by transaminases increase. The up-regulations of SCs markers Cd34 and Sca-1 started from the beginning of the inflammatory stage while progressive increase of Krt19 and Sox9 and CSCs markers Epcam and Cd133 from early hepatic injury. The expressions of Cd133, Cd34, and Afp were significantly higher in HCC compared to paired non-HCC tissue, in contrast to Epcam and Krt19. Western blot and IHC confirmed the positivity of Cd34 and Cd133 in small cells subpopulation

Detection of MBL-2 gene expression in intestinal biopsies of celiac patients by in situ reverse transcription polymerase chain reaction.

Celiac disease (CD) is an autoimmune enteropathy triggered by ingestion of gluten in genetically susceptible subjects and represents one of the most frequently occurring, treatable, lifelong autoimmune disorders. Undetected or untreated CD may cause late more severe complications (Farrell and Kelly, 2002). So far, several factors have been identified as possible agents responsible for CD. There is a strong evidence that CD is associated with specific HLA haplotypes (HLADQA1* 0501, DQB1*0201 or DQA1*0301, DQB0302) (Sollid and Thorsby, 1993). Recently it has been demonstrated on Italian patients that polymorphisms of the first exon of MBL2 gene, which encodes for Mannose Binding Protein (MBP), could play a pathophysiological role in celiac disease (Boniotto et al., 2002). MBP is a serum protein involved in the natural or innate immune response. MBP acts as an ante-antibody and can enhance opsonisation, or can activate the classical pathway of the complement on bacteria, viruses and fungi (Sastry and Ezekowitz, 1993)

MBL Interferes with Endovascular Trophoblast Invasion in Pre-Eclampsia

The spiral arteries undergo physiologic changes during pregnancy, and the failure of this process may lead to a spectrum of pregnancy disorders, including pre-eclampsia. Our recent data indicate that decidual endothelial cells (DECs), covering the inner side of the spiral arteries, acquire the ability to synthesize C1q, which acts as a link between endovascular trophoblast and DECs favouring the process of vascular remodelling. In this study, we have shown that sera obtained from pre-eclamptic patients strongly inhibit the interaction between extravillous trophoblast (EVT) and DECs, preventing endovascular invasion of trophoblast cells. We further demonstrated that mannose-binding lectin (MBL), one of the factor increased in pre-eclamptic patient sera, strongly inhibits the interaction of EVT with C1q interfering with the process of EVT adhesion to and migration through DECs. These data suggest that the increased level of MBL in pre-eclampsia may contribute to the failure of the endovascular invasion of trophoblast cells

Candidate Biomarkers for the Detection of Serious Infections in Children: A Prospective Clinical Study

Serious bacterial infections (SBI) in children are associated with considerable morbidity and mortality, and their early identification remains challenging. The role of laboratory tests in this setting is still debated, and new biomarkers are needed. This prospective, observational, single-center study aims to evaluate the diagnostic role of blood biomarkers in detecting SBI in children presenting with signs of systemic inflammatory response syndrome (SIRS). A panel of biomarkers was performed, including C-reactive protein (CRP), procalcitonin (PCT), white blood cell count (WBC), absolute neutrophil count (ANC), interleukin (IL)-6, IL-8, IL-10, human terminal complement complex (C5b-9), Plasmalemma-Vesicle-associated protein 1 (PV-1), Intercellular Adhesion Molecule-1 (ICAM-1), and Phospholipase A2 (PLA2). Among 103 patients (median age 2.9 years, 60% males), 39 had a diagnosis of SBI (38%). Significant predictors of SBI were CRP (p = 0.001) and ICAM-1 (p = 0.043). WBC (p = 0.035), ANC (p = 0.012) and ANC/WBC ratio (p = 0.015) were also significantly associated with SBI in children without pre-existing neutropenia. ROC curves, however, revealed suboptimal performance for all variables. Nevertheless, a model that combined CRP and ANC/WBC ratio had more in-depth diagnostic accuracy than either of the two variables. Overall, this study confirms the limited usefulness of blood biomarkers for the early diagnosis of SBI. WBC, ANC, ANC/WBC ratio, CRP, and ICAM-1 showed the best, albeit moderate, diagnostic accuracy

Circulating TRAIL Shows a Significant Post-Partum Decline Associated to Stressful Conditions

Background: Since circulating levels of TNF-related apoptosis inducing ligand (TRAIL) may be important in the physiopathology of pregnancy, we tested the hypothesis that TRAIL levels change at delivery in response to stressful conditions. Methods/Principal Findings: We conducted a longitudinal study in a cohort of 73 women examined at week 12, week 16, delivery and in the corresponding cord blood (CB). Serum TRAIL was assessed in relationship with maternal characteristics and to biochemical parameters. TRAIL did not vary between 12 (67.6627.6 pg/ml, means6SD) and 16 (64.0616.2 pg/ml) weeks ’ gestation, while displaying a significant decline after partum (49.3626.4 pg/ml). Using a cut-off decline.20 pg/ml between week 12 and delivery, the subset of women with the higher decline of circulating TRAIL (41.7%) showed the following characteristics: i) nullipara, ii) higher age, iii) operational vaginal delivery or urgent CS, iv) did not receive analgesia during labor, v) induced labor. CB TRAIL was significantly higher (131.6652 pg/ml) with respect to the corresponding maternal TRAIL, and the variables significantly associated with the first quartile of CB TRAIL (,90 pg/ml) were higher prepregnancy BMI, induction of labor and fetal distress. With respect to the biochemical parameters, maternal TRAIL at delivery showed an inverse correlation with C-reactive protein (CRP), total cortisol, glycemia and insulin at bivariate analysis, but only with CRP at multivariate analysis

Association between up-regulated expression proteins and circulating steroidal hormones in leiomyoma

Several studies have shown that both ovarian hormones estrogen and progesterone and various proteins are involved in uterine leiomyoma growth. We hereby describe a study carried out to investigate the possible relation between progesterone, estrone, 17\u3b2-estradiol and the seven dysregulated proteins in the leiomyoma. Statistical analyses showed a significant inverse rank correlation between desmin expressed in leiomyoma and progesterone (\u3c1 = 120.9276; p = 0.0077) and between alpha-1-antitrypsin expressed in leiomyoma and 17\u3b2-estradiol (\u3c1 = 120.8571; p = 0.0137). Our data suggest that decreased levels of 17-\u3b2-estradiol involve an increasing of the inflammation response stimulating alpha-1-antitrypsin expression in leiomyoma and that lower levels of progesterone associated with increasing of desmin expression may be related to increase of inflammation response, and to the role played by desmin in signal transduction inducing leiomyoma growth

Analysis of the cAMP-PCT-CGRP axis in normal and pathological pregnancy

The pro-hormone Procalcitonin (PCT) is one of the best diagnostic and prognostic markers in clinical practice, which exerts an immunomodulatory role during inflammation. We previously showed that PCT has turned out to be a powerful tool to study the ability of pregnant serum to regulate mononuclear phagocytes function. The modulation of PCT may help us to understand the mechanisms responsible for the imbalance in the immune-endocrine crosstalk in pregnancy and encourage further research to identify effective therapeutic strategies in pre-eclampsia (PE). Our recent findings show that PCT levels are higher in PE sera compared to those of healthy pregnant women. We demonstrated the local synthesis of PCT in normal and PE placenta due to macrophages and trophoblast cells. PE sera have the ability to upregulate the PCT production in trophoblasts cells and M\u3a6. TNF\u3b1, IL-1\u3b2 and IL-6 are more concentrated in PE serum, their expression is upregulated in M1 M\u3a6, and seem to drive together the regulation of the synthesis of PCT and CGRP. We show that TNF\u3b1 alone is not able to increase the PCT expression in human M\u3a6. A mAb to TNF\u3b1 completely abrogated the ability of PE sera to upregulate PCT production in these cells. These data emphasize that additional factors in the gravid serum play a role in the PCT synthesis, indicating that its regulation might have an impact on the use of this laboratory marker for the analysis of the inflammatory status in healthy and pathological pregnancy

Predittivit\ue0 dell'emoglobina glicata nei confronti dell'OGTT in una popolazione pediatrica italiana sovrappeso/obesa

Background. Recently glycated hemoglobin (A1c) 65 48 mmol/mol has been included among the diagnostic criteria for type 2 diabetes mellitus (DM2) in adults, while borderline levels of 39-47 mmol/mol indicate impaired glycemia (pre-diabetes). In overweight/ obese children an oral glucose tolerance test (OGTT) is used to screen for DM2 and pre-diabetic status (impaired fasting glucose [IFG] and impaired glucose tolerance [IGT]). A1c could therefore theoretically replace OGTT as a screening test in children at risk for DM2, IFG, IGT, or insulin resistance (IR). Methods. Italian patients aged less than 18 years, consecutively assessed for overweight/obesity (BMI > 85th percentile) in our Pediatric Endocrinology Outpatient Care Unit between July 2011 and December 2012, were included. A1c was determined for each patient (IFCC method) and an OGTT was done (blood glucose and insulin levels at 0, 60 and 120 min). Results. Data were collected from 225 patients (105 males), mean age 11.2 \ub1 3.2 years and mean BMI 27.6 \ub1 4.7 (77% with BMI > 95th percentile). Mean A1c was 36.2 \ub1 4.9 mmol/mol (range 16-50). Three cases had IFG, 12 IGT and 83 IR. In 71 cases (32%) with borderline A1c, none had IFG, 5 had IGT, and 37 had IR. The only child known to be diabetic had borderline A1c. Sensitivity and specificity were 0, 67.6% for IFG, 41.6, 68.8% for IGT, 45.1, 76.1% for IR. Conclusions. A1c is not sensitive enough for detecting diabetic and pre-diabetic status, so it would not be a reliable tool. Given the intermediate specificity of borderline A1c 39-47 mmol/mol to identify pre-diabetic status, confirmation with an OGTT is always advisable. In our series, borderline A1c was better at predicting IR

Analysis of the molecular mechanisms of C1-inhibitor deficiency induced angioedema

Purpose: Angioedema (AE) due to inherited or acquired deficiencies of C1 inhibitor (C1-INH) is characterized by localized swelling of deeper layers of the skin or submucosal tissues, becoming particularly life threatening if it occurs in the upper respiratory tract. C1-INH regulates the release of bradykinin which can enhance permeability of post-capillary venules interacting with its receptors. The drugs currently used are more symptomatic than curative, so we sought to identify the molecular mechanisms responsible for the induction of vascular permeability. Methods: We used a transwell in vitro model with a filter covered by primary human endothelial cells (EC), in the upper chamber we add the fluorescent-BSA and the stimuli and the BSA leaked into the lower chamber was evaluated using a Fluorescence reader. Results: EC were incubated with plasma collected from patients during attack (APL) and the presence of C1-INH in the majority of the patients was able to block the permeability. To mimic the in vivo situation we stimulated the EC with the APL for 30 min and then the SN was collected and used to stimulate the ECs in the transwell model. In that case the inhibition of the leakage by C1-INH was not seen in all the patients. This observation was further confirmed by using the plasma collected from 1 patient before and 1 h after the clinical treatment with C1-INH, indeed there is no difference in the EC leakage induced by the plasma before and after the treatment. Discussions: The inhibition of endothelial leakage induced by APL stimulation by C1-INH indicates the involvement of that molecule in controlling the onset of AE attacks, although the inability of C1-INH to completely block the permeabilizing effect of the SN indicates that after the activation of the cells there are other molecules involved. Conclusion: Since the clinical treatment of AE can be done with different drugs besides C1-INH we have to analyze the most appropriate therapeutic approach