The Role of Neutrophil Extracellular Traps in the Outcome of Malignant Epitheliomas: Significance of CA215 Involvement.

Neutrophil extracellular traps (NETs) were originally discovered as a part of the innate immune response of the host to bacteria. They form a web-like structure that can immobilize microorganisms or exhibit direct antimicrobial properties, such as releasing reactive oxygen species (ROS). NETs are established when neutrophils undergo a sort of cellular death following exposure to ROS, chemokines, cytokines, or other soluble factors. This process results in the release of the neutrophil's DNA in a web-like form, which is decorated with citrullinated histones (H3/H4-cit), neutrophil elastase (NE), and myeloperoxidase (MPO). Emerging studies have put into perspective that NETs play an important role in oncology as they were shown to influence tumor growth, malignant initiation, and proliferation, mediate the transition from endothelial to mesenchymal tissue, stimulate angiogenesis or metastasis, and can even help cancer cells evade the immune response. The role of NETs in cancer therapy resides in their ability to form and act as a mechanical barrier that will provide the primary tumor with a reduced response to irradiation or pharmaceutical penetration. Subsequently, cancer cells are shown to internalize NETs and use them as a strong antioxidant when pharmaceutical treatment is administered. In this review, we explored the role of NETs as part of the tumor microenvironment (TME), in the context of malignant epitheliomas, which are capable of an autonomous production of CA215, a subvariant of IgG, and part of the carcinoembryonic antigen (CEA) superfamily. Studies have shown that CA215 has a functional Fc subdivision able to activate the Fc-gamma-RS receptor on the surface of neutrophils. This activation may afterward stimulate the production of NETs, thus indicating CA215 as a potential factor in cancer therapy surveillance.

Continued benefit demonstrated with BAY 81-8973 prophylaxis in previously treated children with severe haemophilia A: Interim analysis from the LEOPOLD Kids extension study.

INTRODUCTION: BAY 81-8973 (Kovaltry®), a recombinant factor VIII (rFVIII) product, was efficacious and well tolerated in paediatric previously treated patients (PTPs) with severe haemophilia A for ≥50 exposure days (EDs) in the LEOPOLD Kids study. Because long-term prophylaxis (≥100 EDs) can provide substantial patient benefits, FVIII products should demonstrate long-term safety and efficacy. AIM: To demonstrate long-term (≥100 EDs) efficacy and safety of BAY 81-8973 in paediatric PTPs. METHODS: PTPs aged ≤12 years with severe haemophilia A without inhibitors could continue in the ongoing open-label extension study after completing ≥50 EDs in the LEOPOLD Kids main study. Patients received BAY 81-8973 for prophylaxis (25-50 IU/kg ≥2×/week), bleed treatment, and surgery. Bleeds were documented in electronic patient diaries. Inhibitor development was monitored every 6 months. RESULTS: At the August 2017 interim data cutoff, 46 patients (median [range] age at enrolment, 6.0 [1.0-11.0] years) had spent a median (range) of 602.5 (148-1069) EDs and 4.6 (1.0-5.9) years in the main plus extension studies. Median (quartile [Q]1; Q3) annualised bleeding rate for bleeds within 48 h after a prophylaxis infusion and total bleeds was 1.0 (0.2; 1.9) and 2.0 (0.4; 3.6), respectively. Most (>94%) bleeds were mild or moderate; 71.8% were treated with ≤1 infusion. BAY 81-8973 was also well tolerated with only one treatment-related adverse event (transient, low-titre inhibitor which did not require treatment adjustment). CONCLUSION: BAY 81-8973 was efficacious for prophylaxis and treatment of bleeds during >4.5 years in paediatric PTPs with severe haemophilia A.

Influence of chronic administration of anabolic androgenic steroids and taurine on haemostasis profile in rats: a thrombelastographic study.

Anabolic androgenic steroids (AAS) are synthetic derivatives of testosterone with thrombogenic potential in high doses and long-term administration. Taurine, a widely distributed amino-sulfonic acid, is known for its beneficial effects in hypercoagulable states. In order to assess the impact of chronic administration of high doses of AAS and taurine upon haemostasis process in rats, 40 male Wistar rats were divided into four equal groups: control group (group C) - no treatment; androgen group (group A) - received 10 mg/kg per week of nandrolone decanoate (DECA); taurine (group T) - received oral supplementation of 2% taurine in drinking water; androgen and taurine group (group AT) - concomitant administration of DECA and taurine. After 12 weeks, blood samples were collected and haemostasis parameters were assessed with the thrombelastographic (TEG) analysis system: reaction time, clot kinetics (K, α), final clot strength, coagulation index and the clot lysis (Ly30). Nandrolone significantly decreased reaction time in group A compared with control (P<0.001), whereas taurine significantly increase reaction time (P=0.01), and this effect was maintained in group AT compared with group A (P=0.009). Similar differences between groups have been recorded for the clot kinetics parameters K, α. The final clot strength and coagulation index were significantly increased in group A versus group C (P=0.04, respectively P<0.001), but not in group AT versus group C (P>0.05). There were no differences in clot lysis, as shown by Ly30. Nandrolone produces an accelerated clot development and an increased clot firmness in Wistar rats. Taurine association ensures a protective effect against this hypercoagulable state, partially restoring the altered parameters of the coagulation profile.

Interleukin-6 and interleukin-10 gene polymorphism, endothelial dysfunction, and postoperative prognosis in patients with peripheral arterial disease.

OBJECTIVES: The aim of this study was to evaluate the association between interleukin (IL)-6 and IL-10 gene polymorphism and the short-term risk of postoperative cardiovascular events in patients with peripheral artery disease receiving elective surgery and also to evaluate the endothelial function. METHODS AND RESULTS: We determined preoperatively IL-6 gene polymorphism (-174 G/C and nt565 G/A), IL-10 polymorphism (-1082G/A, -819C/T, -592C/A), and brachial artery vasodilatation using ultrasound in 48 patients undergoing vascular surgery. Eight patients (16.7%) developed over a period of 30 days cardiovascular events (cardiovascular death, resuscitated cardiac arrest, acute myocardial infarction, unstable angina, stroke). Cardiovascular events were more frequent in the subgroups of patients with genotypes associated with high serum levels of IL-6: -174CC (57.14% vs 12.5% for -174GC genotype and 8% for -174GG, P = .007) and nt565AA (50% vs 17.6% for nt565GA genotype and 8% for nt565GG genotype, P = .021) and in subgroups with haplotypes associated with low serum levels of IL-10: ATA (57.14% vs 14.8% for haplotype ACC and 7.4% for GCC, GCA, GTA, GTC haplotypes, P = .004). Flow-mediated dilatation was significantly lower in patients with IL-6 -174CC genotype (7.05% +/- 1.49% vs 8.41% +/- 1.9% for IL-6 -174GC and 9.42% +/- 2.46% for IL-6 -174GG, P = .009) and IL-6 nt565AA genotype (7.14 +/- 1.61% vs 8.49% +/- 1.91% for IL-6 nt565GA and 9.42% +/- 2.46% for IL-6 nt565GG, P = .018) and in patients with IL-10ATA haplotype (6.45% +/- 0.57% vs 9.13% +/- 2.52% for IL-10ACC and 9.24% +/- 2.09% for IL-10 GCC/GCA/GTA/GTC, P = .004) respectively. CONCLUSIONS: IL-6 -174CC and nt565AA genotypes and IL-10ATA haplotypes are correlated with a high short-term risk of acute postoperative cardiovascular events in patients with peripheral artery disease receiving elective surgical revascularization and with endothelial dysfunction in these patients.

In vitro hepatic differentiation of human bone marrow mesenchymal stem cells under differential exposure to liver-specific factors.

Recent findings demonstrated that stem cells could be harvested from a patient and used to repair his or her own damaged liver. Additionally, stem cells may be manipulated in vitro to induce hepatic differentiation. The current study aims to determine the differentiation efficacy of various liver-specific factors (hepatocyte growth factor, Insulin-Transferrin-Selenium, dexamethasone, and nicotinamide) used for stem cell differentiation into hepatocyte-like cells. Human mesenchymal stem cells were exposed to different media containing these compounds added individually or in various combinations. Hepatic differentiation was assessed via quantitative reverse transcription-polymerase chain reaction and immunocytochemical staining for stemness or liver-specific genes and proteins, including albumin, cytokeratins 18 and 19, HepPar-1, alpha-fetoprotein, and nestin. In addition, functional tests for glycogen storage, urea production, glucose, and albumin synthesis were also performed. The expression profiles of albumin, alpha-fetoprotein, and cytokeratin 19 demonstrated that when hepatocyte growth factor, nicotinamide, or dexamethasone were added individually, incomplete hepatocyte differentiation was achieved; the obtained cell populations contained progenitors that expressed both hepatic (albumin) and biliary (cytokeratin 19) markers, as well as alpha-fetoprotein. Hepatocyte growth factor and nicotinamide were the factors with the most hepatogenic potential. When all factors were added to the culture, cells exhibited features that closely resembled human adult hepatocytes as determined by their gene expression patterns (albumin, HepPar-1, and alpha-fetoprotein, but not cytokeratin 19) and functional testing. These cells with hepatic function may become important tools for liver transplant procedures, liver development studies, and pharmacologic research.

Current challenges of anticoagulant treatment.

Anticoagulant therapy plays an important role in current medical practice. The main types of anticoagulant agents are: heparins, hirudins and vitamin K antagonists. None of the drugs used as anticoagulants meet the criteria of an ideal anticoagulant because they have side effects and they interact with other compounds. The main side effect of anticoagulant therapy is bleeding. The choice of a certain anticoagulant is made by the doctor based on the clinical context and also on the desired effect.

Is interleukin-18 correlated with endothelial dysfunction and platelet activation in patients with unstable angina?

UNLABELLED: The aim of the study was to assess the interaction between interleukin-18 (IL-18, a cytokine with a central role in the inflammatory cascade) and the presence of endothelial dysfunction and prothrombotic profile in patients with unstable angina and proven coronary artery disease. MATERIAL AND METHODS: The study EDIT-ACS (Endothelial Dysfunction, Inflammation and Thrombosis in Acute Coronary Syndromes) was set in the Cardiology Department of our Institute, and inclusions were performed between June-November 2004. We included 40 consecutive patients with unstable angina hospitalized in our Department (mean age = 60.2 +/- 8.8 years, 57.5% male), as well as 20 normal pts as a control population,matched for age and sex (mean age = 61.9 +/- 8.6 years, 58.3% male). All pts had the following biological studies performed: interleukin-18 (IL-18), C-reactive protein (CRP), fibrinogen, leukocyte count, and Von Willebrand factor, as well as platelet activity parameters (mean platelet volume, MPV and platelet distribution width, PDW). All pts had echographic studies of flow mediated dilatation (FMD) and measurement of carotid intima-media thickness (C-IMT), and underwent coronary angiography as well. RESULTS: We found that patients in the highest quartile of IL-18 had higher levels of CRP (11.9 +/- 4.2 vs 6.8 +/- 3.4 mg/l, p<0.01), von Willebrand factor (166% vs 150%, p=0.08), PDW (14.5 vs 13.2, p=0.04), MPV (11.2 vs 10.2, p=0.02), as well as a higher number of coronary stenoses (mean no. of affected vessels 2.5 vs 1.1, p<0.001), trend towards higher carotid IMT (0.92 mm vs 0.81 mm, p=0.06). They also have significant endothelial dysfunction (FMD of 5 % vs 8.5% for 4th quartile and 1st quartile respectively). Pts with higher levels of IL-18 associated a worse short term (in hospital prognosis) with 60% of pts in the 4th quartile having adverse events, versus 22% in the lowest quartile. CONCLUSIONS: High levels of interkleukin-18 are associated with endothelial dysfunction (high von Willebrand factor and low brachial FMD) and platelet activation (high MPV and PDW values), in pts with a higher number of affected coronary arteries but similar global atherosclerotic burden. This could explain the association between high levels of IL-18 and worse short-term prognosis in pts with unstable angina.

Anticardiolipin antibodies, recurrent stroke and vascular events: a prospective study on 210 patients.

The presence of anticadiolipin antibodies (aCL) has been associated with vascular occlusive events. However, the role of aCL in predicting ischemic events, particularly ischemic stroke, is controversial. In order to determine if aCL is either associated with thrombotic disease or with an increased risk of recurrent stroke or thrombotic events, we performed a prospective study on a cohort of Romanian patients with stroke that were tested for aCL. We studied the prevalence of cardiovascular risk factors in the two subgroups (aCL positive or aCL negative) considering the age of patients and the medical evolution in the first 12 months after the index stroke. Globally, neither the antecedents of thrombotic events or abortions, nor the clinical or biological profile or the recurrence of vascular events showed any significant difference between the group with or without aCL. Only in two patients the criteria for antiphospholipid syndrome were fulfilled. In 37% of cases, there was a variability of aCL positive titers at 8 weeks after the index event. This raised the necessity to isolate a subpopulation with higher risk of ischemia in the presence of aCL, and the need of more specific subtypes of antiphospholipid antibodies as a marker of thrombophilia.

C-reactive protein, prothrombotic imbalance and endothelial dysfunction in acute coronary syndromes without ST elevation.

UNLABELLED: Inflammation is considered a crucial step in the pathogenesis of acute coronary syndromes (ACS). C-reactive protein (CRP) is proposed to be included in risk stratification of ACS patients. However, it is not yet known if CRP is only a risk marker, or merely a risk factor in the development of ACS. Our study looked at the links between inflammation and the prothrombotic factors present in patients with ACS without ST elevation. MATERIAL AND METHODS: 86 patients (pts), 46 men (53.4%), mean age = 58.2+/-12.4 years-old, with acute coronary syndromes (unstable angina and NSTEMI). The following parameters were measured in all pts on admission: CRP, fibrinogen, blood white cell count, and coagulation parameters: coagulation factor V and VIII, von Willebrand factor (vWf), antithrombin III (AT III), D-dimers (DD), C and S proteins. RESULTS: Mean CRP in the study group was 22.42+/-19.81 mg/dl (limits 1.40-88.8 mg/dl). We worked with quartiles of CRP plasmatic levels, in order to see how magnitude of inflammation correlates with different coagulation and fibrinolysis parameters. When comparing the 1st with the 4th CRP quartiles, we noted that important inflammation (4th quartile) was associated with higher factor von Willebrand (141.3 vs 108.9%, p<0.05), factor 5 (127.5% vs 88%, p<0.01), factor 8 (121.5 vs 117.1%, p=0.04), lower AT III (101.6 vs 118.2%, p<0.05), lower protein C and S. The associations did not keep for PAI-I or D-dimers, which might be associated with the lack of sensibility of fibrinolysis markers in the early period after thrombosis. CONCLUSIONS: Inflammation, as quantified by CRP, appears to be associated with a significant prothrombotic status and endothelial dysfunction (as reflected by high von Willebrand factor).

Study of total homocyst(e)ine levels in type 2 diabetic patients with silent brain infarction.

Hyperhomocysteinemia is thought to have an important role in the pathogenesis of ischemic cerebral infarction. When associated with diabetes mellitus, it might worsen the neurologic course. The aim of the study was to clarify the relation between plasma homocysteine (Hcy) concentrations and silent brain infarction (SBI) in patients with type 2 diabetes mellitus. Total plasma Hcy levels were prospectively studied in 46 patients with type 2 diabetes and SBI (group I), mean age 56+/-5.4 years, as compared to 38 diabetic patients without SBI (group II) and with 31 controls (group III). Homocysteine concentrations were determined using a high-performance liquid chromatography assay. The results were compared using the Student's t test. The mean level of Hcy was 22.6+/-2.4 micromol/l in group I, 19.7+/-1.6 micromol/l in group II and 11.4+/-1.4 micromol/l in group III; between group I and group II p < or = 0.001. These data are consistent with increased Hcy levels in type 2 diabetic patients, contributing to the onset of SBI in some patients. The phenomenon should be considered in any future strategy for the therapy of hyperhomocyst(e)inemia (HHcy).