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Histopathological changes in calcific aortic stenosis (CAS) resemble changes in coronary atherosclerosis. Concerning recent evidence on dietary and gut microbiota-related metabolites representing players in atherosclerosis, we aimed to investigate the link between dietary and gut microbiota-derived metabolites and CAS.We consecutively recruited eligible subjects with moderate-severe CAS (Prevalence of traditional cardiovascular risk factors or co-morbidities did not differ among groups (allWe found a significant association between choline levels and CAS presence and severity depicted on imaging modalities and histopathological examinations. Our study may open new horizons for prevention of CAS.

WOS: 000488637800004, PubMed: 31582678, Objective: Atrial structural remodeling has been suggested to contribute to atrial fibrillation (AF) recurrence following direct-current cardioversion (DCCV). The role of several inflammatory and extracellular matrix turnover markers in AF recurrence following DCCV has been investigated. However, data on the impact of galectin-3, which is known to play a role in various fibrotic conditions, including cardiac fibrosis are lacking. The aim of this study was to demonstrate the predictive role of serum galectin-3 levels in AF recurrence following successful DCCV. Methods: A total of 90 persistent AF patients who were scheduled for DCCV were prospectively enrolled. Serum samples were assayed to determine pre-DCCV galectin-3 levels using the enzyme-linked immunosorbent assay method. Patients were followed up for 3 months for AF recurrence. Results: Of 90 persistent AF patients (mean age: 55.33 +/- 7.94 years; 53.33% male) who underwent successful DCCV, 28 (31.11%) experienced early AF recurrence within 3 months. Patients with AF recurrence had a greater left atrial volume index (LAVI) (33.35 +/- 2.45 mL/m(2) vs. 29.21 +/- 3.08 mL/m(2); p, Hacettepe University Scientific Research Projects Coordination UnitHacettepe University [1993], This project was funded by Hacettepe University Scientific Research Projects Coordination Unit (project number: 1993).

WOS: 000482646600008, PubMed: 31375653, Objective: A well-developed coronary collateral circulation lowers both in-hospital and long-term morbidity and mortality limiting the infarct. Angiogenin (AGN) and osteopontin (OPN) are known to be potent inducers of angiogenesis. The aim of the present study was to investigate the relationship between serum ANG and OPN levels and collateral filling grade in subjects with stable coronary artery disease (SCAD). Methods: A total of 122 age- and gender-matched consecutive patients who were found to have total occlusion (n=70) and no significant stenosis in epicardial coronary arteries (n=52) who underwent coronary angiography due to SCAD between January 2015 and July 2017 were included in the study. AGN and OPN levels were measured using enzyme-linked immunosorbent assay. Coronary collateral circulation was graded using Rentrop's classification of collateral filling. Results: A total of 52 patients (61.60 +/- 11.78 years, 61.5% male) without significant epicardial coronary artery stenosis and 70 patients (62.87 +/- 8.24 years, 65.7% male) with totally occluded coronary arteries were included in the study. Subjects with total occlusion had significantly higher levels of AGN [122.00 (79.00-623.00) pg/mL vs. 98.00 (18.00-160.00) pg/mL, p, Hacettepe University Scientific Research Projects Coordination UnitHacettepe University [845], This research was supported by Hacettepe University Scientific Research Projects Coordination Unit (project no.: 845).

Fibroblasts turn into cancer associated fibroblasts (CAFs) in the tumour microenvironment. CAFs have recently attracted attention for their function as a regulator of immune cell recruitment and function in addition to their tumour-promoting roles. In this study, we aimed to determine the role of CAFs on monocyte recruitment and macrophage polarization in breast cancer. CAFs, which were α-SMA expressing fibroblasts in contrast to normal fibroblasts (NFs), effectively recruited monocytes. Recruitment of monocytes by CAFs might be mediated by monocyte chemotactic protein-1 (MCP-1) as well as stromal cell-derived factor-1 (SDF-1) cytokines. CAFs differentiated the recruited monocytes into M2-like macrophages which are capable of exerting their immunosuppressive roles via the PD-1 axis. CAF-educated monocytes exhibited strong immune suppression unlike NF-educated monocytes and enhanced the motility/invasion of breast cancer cells in addition to increasing the expressions of epithelial–mesenchymal transition (EMT)-related genes and vimentin protein in cancer cells. CAF-educated M1 macrophages displayed increased expression of M2 markers and production of anti-inflammatory cytokine IL-10 in contrast to decreased production of pro-inflammatory cytokine IL-12 compared with control M1 macrophages; suggesting that CAFs were also able to induce the trans-differentiation of M1 macrophages to M2 macrophages. We then investigated the relationship between the infiltration of CAFs and tumour associated macrophages (TAMs) using tissue samples obtained from breast cancer patients. High grade of CAFs significantly correlated with the number of TAMs in human breast cancer tissue samples. It was also associated with higher Ki-67 proliferation index, and higher tumour volume. This result is in line with our finding of increased breast cancer cell proliferation due to the effects of CAF-educated monocytes in vitro. Our results concluded that CAFs play pivotal roles in sculpturing the tumour microenvironment in breast cancer, and therapeutic strategies to reverse the CAF-mediated immunosuppressive microenvironment should be taken into consideration.

Beta‐trace protein (BTP) has emerged as a novel biomarker of cardiovascular risk. In this study, the authors aimed to assess the relationship between BTP levels and presence of atrial fibrillation in patients who had controlled hypertension (HTN) and normal renal function. A total of 80 controlled HTN patients with paroxysmal atrial fibrillation (PAF) and 80 age‐ and sex‐matched controls with controlled HTN were enrolled. Serum BTP levels were measured by enzyme‐linked immunosorbent assay. BTP levels were found to be significantly higher in patients with PAF (P

Background Atrial fibrillation(AF) is the most common sustained arrhythmia. Its most feared sequelae are stroke and peripheral thromboembolism due to atrial thrombi formation. Mechanisms underlying the relationship between platelet activation and left atrial thrombi have not been clearly elucidated yet. We aimed to investigate whether immune-mediated platelet activation occurred in AF patients in this cross-sectional study. Methods Persistent and paroxysmal AF patients who underwent cryoballoon-based AF ablation between March 2015 and July 2016 were included as the patient group. Patients without AF in whom transseptal puncture was performed at the same period for purposes other than AF ablation were included as the control group. Peripheral and left atrial blood samples were obtained for determination of platelet Toll-like receptor(TLR)-2, TLR-4 and high mobility group box-1(HMGB-1) expression levels. Results A total of 75 subjects (53 patients with AF and 22 control subjects) [mean: 60.33 (SD: 6.14) years, 57.33% male] were included. Left atrial and peripheral TLR-2, 4 and HMGB-1 expression levels were significantly higher in the patient group when compared to the controls. Left atrial platelet TLR-2 and TLR-4 expression and serum HMGB-1 levels were higher in persistent AF patients compared to paroxysmal AF patients. In the patient group, left atrial expression of TLR-2, 4 and HMGB-1 were significantly higher than the peripheral expression levels. Conclusion Findings of our study suggest evidence for immune-mediated platelet activation in the left atria of AF patients.

WOS: 000416049100009, PubMed: 28295609, BackgroundLeft atrial appendage flow velocity (LAAFV) and presence of spontaneous echo contrast (SEC) have been reported to be predictors of thromboembolism in atrial fibrillation (AF) patients. Galectin-3 is a biomarker reflecting pro-inflammatory status, whose role in AF has recently drawn attention, particularly in persistent AF population. AimIn this study we aimed to investigate the association between serum galectin-3 levels and echocardiographic predictors of thromboembolism in persistent AF patients. MethodsWe included 65 persistent AF patients (55.5010.67years, 46.15% male). Transesophageal echocardiography (TEE) was performed to assess LAAFV and presence of left atrial (LA)/LA appendage (LAA)-located SEC and thrombus prior to direct current cardioversion or catheter ablation for AF. ResultsMedian galectin-3 level was 0.63ng/mL. Serum galectin-3 levels were significantly correlated with LAAFV (r=-.440, P, Hacettepe University Scientific Research Projects Coordination UnitHacettepe University [1993], This research was supported by Hacettepe University Scientific Research Projects Coordination Unit (project number: 1993).

Pathophysiologic mechanisms underlying lone atrial fibrillation (AF) have not been clearly demonstrated yet. Emerging evidence has indicated that autoimmunity may play a role in the development of AF. Relationship between serum anti-M2-muscarinic receptor autoantibody (anti-M2-R) and anti-β1-adrenergic receptor autoantibody (anti-β1-R) levels and lone paroxysmal atrial fibrillation (PAF) has not been investigated. We aimed to compare anti-M2-R and anti-β1-R levels between lone PAF patients and healthy control subjects. 75 patients with lone PAF (age: 52.80 ± 6.80 years, 53 % male) and 75 healthy control subjects (age: 53.30 ± 6.80 years, 54 % male) were enrolled in the study. Serum anti-M2-R and anti-β1-R levels were measured by ELISA and compared between two groups. Anti-M2-R [142.30 (77.65–400.00) vs. 69.00 (39.48–299.04) ng/mL; p

Granulocyte-colony stimulating factor (G-CSF) has become the most effective agent supporting hematopoietic stem cell transplantation (HSCT). The cognate interaction between G-CSF and its specific receptor, G-CSFR, induces the mobilization of HSCs and increases their pool in the peripheral blood. G-CSFR has a highly conserved structure which may be functionally modulated by the presence of missense single nucleotide polymorphisms (SNPs). In this study, we asked whether the missense SNPs in G-CSFR could affect the response to G-CSF in HSCT patients and donors. Here, for the first time, G-CSFR missense SNPs were screened and minor allele frequencies were determined in a specific population with Turkish racial background. Five (rs3917991, rs3918001, rs3918018, rs3918019, and rs146617729) out of 16 missense SNPs screened were determined with minor allele frequencies lower than 0.04. Subsequent association analyses indicated potential impact of rs3918001, rs3918018, and rs3918019 minor alleles on peripheral blood CD34+ cell enrichment. Although their frequency is rather low, certain missense SNPs, especially which are placed in the conserved regions of G-CSFR may possess the capacity to influence the response to G-CSF treatment.

Th17-related cytokines (IL-17, IL-23, and IL-26) and receptors (IL-17R and IL-23R) were evaluated in MS patients under immunomodulatory IFN-β1 therapy during a 2year follow-up. Before the initiation of treatment, no significant difference was found in cytokine or receptor expression between controls and MS patients. Of the three cytokines evaluated, IL-26 was the highest in the patients' sera. The amount of IL-17 and CD13(+)IL-17R(+) cells was steadily decreased whereas IL-23 and IL-26 levels were gradually increased with IFN-β1 therapy. The patients in progressive phase had very high levels of IL-17. Th17-associated parameters should be considered in the immunomodulatory IFN-β1 therapy of MS.

Photodynamic therapy (PDT) is a treatment modality for certain malignant (skin, head and neck, gastrointestinal, gynecological cancers), premalignant (actinic keratosis), and nonmalignant (psoriasis) indications. Broader acceptance by the medical community and applicability is hampered, at least in part, by the less than optimal photophysical characteristics of the porphyrin derivatives. This situation sparked a worldwide search for novel sensitizers leading to new compounds, some holding more promise than others. The primary cytotoxic agent involved in the photodynamic action is singlet oxygen (Dg), the efficient generation of which is linked invariably to the intersystem crossing (ISC) efficiency of the excited sensitizer. Most organic dyes have low triplet quantum yields, and in many recent candidates for photodynamic sensitizers, heavy atoms are incorporated into the structure as a strategy to improve spin–orbit coupling leading to facilitated intersystem crossing. While this approach seems fail-safe, incorporation of heavy atoms such as bromine, iodine, selenium, and certain lanthanides very often leads to increased “dark toxicity”. Unlike traditional chemotherapy agents, in principle, photodynamic therapy sensitizers themselves can be nontoxic, either at cellular or organ levels, even at relatively high concentrations. We have been interested in trying to find alternative ways of achieving increased intersystem crossing without the use of heavy atoms to minimize dark toxicity, turning our attention to the excitedstate properties of the sensitizers. Designing efficient photoinduced O2 generators requires that any existing operative fluorescence cycle of the fluorophore, which is through the S0!S1!S0 states, has to be perturbed so as to minimize or shut down the S1!S0 deactivation, and switch to the triplet surface once S1 is accessed. A general design principle for a favorable S1!T1 transition from an electronic structure viewpoint would in principle require the structural and electronic compatibility of the S1 and T1 states to surpass that of the S1–S0 pair. Once multiple electronic states come into play, quantum mechanical calculations providing a detailed understanding of the electronic structure are extremely helpful. Multi-configurational self-consistent field (MCSCF) techniques are the stateof-the-art computational chemistry approaches, when near degeneracies and excited states are considered. These methods may not reach chemical accuracy ( 2–3 kcalmol ) for computing total energies, but they are crucial for a qualitatively correct description of the excited states and are capable of providing a conceptually complete picture of the photophysics taking place. Therefore, we mainly employed a popular variant of MCSCF techniques; the complete active space SCF (CASSCF) method in combination with relatively large basis sets and different active spaces. Details of CASSCF calculations are provided in the Supporting Information. Our calculations on the parent Bodipy (4,4-difluoro-4-bora-3a,4adiaza-s-indacene, Scheme 1) showed that natural orbital occupancies of the S1 state describe an open-shell singlet with essentially double (> 1.9) or zero (< 0.1) electrons for all orbitals except the highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) that are singly occupied (see the Supporting Information, Figure S1). It is no surprise to observe a fluorophore with low triplet quantum yield to have an excited state that possesses only two orbitals with single occupancy. Hence, to achieve our goal of efficient switching to the triplet manifold, we have to access excited states that differ from the ones that arise from simple HOMO!LUMO transitions. Among multiply excited configurations, doubly substituted ones are particularly important in enhancing S1–T1 coupling as shown by the seminal work of Salem and Rowland and the following work by Michl. Thus, the substitutions should invoke a simultaneous two-electron excitation from the Scheme 1. Structure and numbering of the parent Bodipy compound. [*] Y. Cakmak, S. Kolemen, B. Kilic, Prof. Dr. E. U. Akkaya UNAM-Institute of Materials Science and Nanotechnology Bilkent University, Ankara, 06800 (Turkey) E-mail: eua@fen.bilkent.edu.tr

The extracellular matrix plays a critical role in macrophage maturation. In this study, the HL60 cell line was used as a model of leukemic myeloid cell differentiation. We assessed the ability of HL60 cells cultured on fibronectin substratum prior to phorbol 12-myristate 13-acetate (PMA) induction to differentiate into terminally differentiated macrophages. Beside their distinctive macrophage morphology, they expressed antigen receptors CD14, TLR2, TLR4 and CD68, and displayed enhanced phagocytic activity and production of reactive oxygen species. Expression of CD13, CD33, CD15 and alpha-naphthyl-acetate esterase was also maintained, however, differentiated HL60 cells were HLA-DR and CD1a negative. Here, we describe the enhanced capacity of fibronectin-adherent HL60 cells to differentiate into macrophages in response to PMA.

Atrial fibrillation (AF) is the most common sustained arrhythmia. Inflammation has been suggested to play a vital role in the pathogenesis. Previous studies have investigated expression of inflammatory markers in AF. Several studies have focused on the effects of toll-like receptors (TLRs) on heart in terms of capability of modulating inflammation. In this study, we aimed to investigate whether peripheral monocyte TLR expression was associated with the AF presence, and recurrence of AF after cryoablation, as a reflection of inflammatory status. Patients with AF who were scheduled for cryoballoon-based ablation for AF and age- and gender-matched subjects in sinus rhythm were included. Peripheral monocyte TLR-2 and TLR-4 expressions were evaluated by flow cytometric analysis in peripheral venous blood samples obtained during evaluation in outpatient clinics: 172 patients (56.5 ± 6.6 years, 52.3% men) were included in the study. Peripheral monocyte TLR-2 and TLR-4 expression levels were significantly higher in patients with AF (p0.05). Among patients with AF, 12 patients (14.0%) developed AF recurrence at a follow- up of 17 months. Multivariate Cox regression analysis showed that left atrial volume index (hazard ratio 2.040, 95% CI 1.197 to 3.477, p = 0.009) and monocyte TLR-4 expression (hazard ratio 1.226, 95% CI 1.042 to 1.443, p = 0.014) were independent predictors of AF recurrence after blanking period following second-generation cryoballoon-based pulmonary vein isolation for paroxysmal AF. In conclusion, our study highlights the role of TLR-mediated inflammation in the pathogenesis of AF. This link may also constitute a therapeutic target in patients with AF.

Purpose Hemophagocytic lymphohistiocytosis (HLH) is a life-threatening condition characterized by uncontrolled inflammation and has common clinical and laboratory features with sepsis. The aim of this study was to investigate patients treated with severe sepsis who had bicytopenia for the presence of HLH. Materials and methods Patients with severe sepsis who were non-responsive to treatment and developed at least bicytopenia were included. Peripheral blood samples were collected and stored for later evaluation for natural killer (NK) activity and soluble interleukin-2 receptor levels. Diagnostic criteria of HLH were retrospectively analyzed. Results Seventy-five of 382 patients (20%) were followed as severe sepsis and septic shock. Among them, 40 patients had bicytopenia. Twenty-six of 40 patients were excluded due to the presence of active solid or hematological malignancies. Three patients died before fulfillment of HLH criteria and one patient denied to give consent. All of the remaining 10 patients had at least five of the eight criteria according to criteria of the Histiocyte Society. Only one of 10 patients was diagnosed as HLH and received treatment during intensive care unit stay. None of the 10 patients survived. Conclusions This study emphasizes to consider the possibility of HLH and the need of rapid assessment of patients with severe sepsis who had bicytopenia and were resistant to treatment in intensive care.

Evidence is accumulating that toll-like receptors (TLR) are involved in the initiation and progression of cardiovascular disease. Enhanced expression of these receptors on monocytes has been shown in patients with acute coronary syndrome (ACS). However, expression on platelets in this group of patients has not been evaluated yet. We aimed to demonstrate the possible relationship of platelet TLR-2 and TLR-4 expressions with stable coronary artery disease and ACS pathogenesis. In this observational case-control study, 40 patients diagnosed with ACS (unstable angina pectoris, non-ST-segment elevation and ST-segment elevation ACS), 40 patients diagnosed with stable coronary artery disease, and 40 age- and gender-matched subjects with normal coronary arteries were involved. Platelet TLR-2 and TLR-4 expressions were evaluated by flow cytometry in peripheral venous blood samples obtained before coronary angiography. A total of 120 patients (60.7 ± 12.3 years, 50% men) were included. Median platelet TLR-2 and TLR-4 expressions were greater in patients with ACS compared to those with stable angina pectoris and normal coronary arteries (29.5% vs 10.5% vs 3.0%, p0.001 and 40.5% vs 11.5% vs 3.0%, p0.001, respectively). Median platelet TLR-2 and TLR-4 expressions were also greater in patients with stable angina pectoris compared to those with normal coronary arteries (p0.05). In conclusion, this is the first study demonstrating enhanced TLR-2 and TLR-4 expressions on platelets in patients with ACS. These findings may suggest that platelet TLR expression as a novel potential prophylactic and therapeutic target in ACS.

Recent evidence has suggested that autoantibodies may play an important role in the development of atrial fibrillation (AF). The predictive value of preprocedural autoantibodies against beta-1 adrenergic receptor (anti-β1-R) and M2-muscarinic acetylcholine receptor (anti-M2-R) for AF recurrence following cryoballoon-based pulmonary vein isolation (PVI) is still unclear. We aimed to determine the predictive value of preprocedural anti-β1-R and anti-M2-R levels for AF recurrence.Eighty patients (mean age 54.25 ± 7.70 years; 40% female) with paroxysmal AF and preserved left ventricular function who underwent cryoballoon-based PVI were included in the study. Preprocedural anti-M2-R and anti-β1-R levels were measured with ELISA.At 1-year follow-up after ablation, late AF recurrence was observed in 17 (21.25%) patients. In the Cox regression model, including number of antiarrhythmic drugs, early AF recurrence, anti-β1-R levels159.88 ng/mL, anti-M2-R levels277.65 ng/mL, AF duration, and left atrial volume index, only anti-β1-R levels159.88 ng/mL (HR: 4.281, P = 0.039) and anti-M2-R levels277.65 ng/mL (HR: 4.313, P = 0.030) were found to be independent predictors of late AF recurrence. Anti-β1-R level159.88 ng/mL was shown to predict late AF recurrence with a sensitivity of 70.59% and specificity of 90.48%. A cut-off value of 277.65 ng/mL for anti-M2-R level predicted AF recurrence with a sensitivity of 70.59% and specificity of 95.24%.Preprocedural serum anti-β1-R and anti-M2-R levels are independent predictors of late AF recurrence following cryoballoon-based PVI in paroxysmal AF patients. Detection of preprocedural anti-β1-R and anti-M2-R levels may serve as a novel method for determination of paroxysmal AF patients who may not benefit from cryoballoon-based PVI.

In the present study, the preparation and characterization of bovine serum albumin (BSA) microspheres and the evaluation of the in vitro cytotoxicity of these microspheres on acute promyelocytic leukaemia (HL-60) cells were described. Mitoxantrone (MTZ)-incorporated microspheres were evaluated for particle size, drug loading, release characteristics and surface morphology. The biological effect of MTZ released from BSA microspheres was determined on an in vitro cultured HL-60 cell line, showing that, after encapsulation, MTZ still retains cytotoxic activity. For this purpose, methyl-thiazol-tetrazolium (MTT) assay was used to evaluate the in vitro cytotoxicity of MTZ-loaded microspheres. Particle size of BSA microspheres was determined between 17.61-20.38 microm and they were smooth and spherical in shape. Encapsulation efficiency of the drug-loaded microspheres was between 22.26-60.50%. For MTZ-containing microspheres, the cell death ratios were greater than 80% for all formulations. This study demonstrate that BSA microspheres were well suited for the controlled release of MTZ and were promising for anti-cancer chemotherapy.

Both atopy and asthma are claimed to be associated with a Th-2 cytokine pattern. We sought to determine the contribution of atopy and asthma to the observed Th-2/Th-1 imbalance in these conditions. Of 60 children aged 6-16 years that were included in the study, 13 were nonatopic nonasthmatic, 15 atopic nonasthmatic, 14 nonatopic asthmatic, and 18 atopic asthmatic. Atopic children had positive skin prick tests to grass pollens only. All children were studied after an asymptomatic and drug-free period of at least three months. Total IgE was measured in serum. Peripheral blood mononuclear cells were cultured and stimulated in vitro with phytohemagglutinin and interferongamma (IFN-gamma) and interleukin-4 (IL-4) measured in the supernatants. Total IgE was significantly higher in atopic asthmatics compared to nonatopic asthmatics (p = 0.004), and nonatopic nonasthmatics (p = 0.001), but was not different from atopic nonasthmatics (p0.05). On the other hand, IL-4 was significantly elevated in atopic asthmatics and in nonatopic asthmatics compared to nonatopic nonasthmatics (p = 0.037 and p = 0.009, respectively). Although atopic asthmatics had lower IFN-gamma values than nonatopic asthmatics, the difference did not reach statistical significance. No correlation was detected between any two parameters. Our results suggest that both atopy and asthma contribute to the increased levels of IL-4 and that, whereas nonatopic asthma is associated with increases in both IL-4 and IFN-gamma release by mononuclear cells, only atopic asthma is characterized by a Th-2 type cytokine dominance.

Fluorouracil has been reported to induce severe side-effects in particular subjects who have deficiency in dehydropyrimidine dehyrogenase activity, the major enzyme in the catabolism of fluorouracil.In this study, we aimed to analyze the heterozygote and homozygote frequencies of dehydropyrimidine dehyrogenase gene mutation in 200 patients receiving fluorouracil based chemotherapy together with the assessment of the toxicity profile of these chemotherapy regimens.According to the results of clinical toxicity assessments, grade 3-4 hematologic toxicity was noted in 12% of the patients. Grade 3 gastrointestinal toxicity was present in 5% of the subjects with no grade 4 side effects. The heterozygote (2q(1-q)) and homozygote (q2) frequencies of dehydropyrimidine dehyrogenase gene mutation were calculated as 1.5% (3/200) and 0.000055% (1/18,043) in the analyzed samples.In this report, for the first time we documented the frequency of dehydropyrimidine dehydrogenase gene mutation in Turkish cancer patients. The determination of enzyme activity in suspected individuals and analysis of other mutations on a population basis would be the next steps for our country.

Galectin-3 is known to play an important role in a number of fibrotic conditions, including cardiac fibrosis. Many studies have focused on the association between galectin-3 levels and cardiac fibrosis in heart failure. However, the role of galectin-3 in the pathogenesis of atrial fibrillation (AF) has not been evaluated thoroughly yet. The aim of this study was to determine whether serum galectin-3 levels were elevated in patients with AF and preserved left ventricular function. Seventy-six patients with paroxysmal or persistent AF and preserved left ventricular systolic function and 75 age- and gender-matched control subjects were enrolled in this observational study. Galectin-3 levels were measured by enzyme-linked immunosorbent assay. Serum galectin-3 (median 0.6 ng/ml [interquartile range 0.2 to 1.4] vs 0.5 ng/ml [interquartile range 0.1 to 0.7], p0.001) and left atrial volume index (LAVI) (mean 29.5 ± 3.5 vs 26.5 ± 2.5 ml/m(2), p0.001) were significantly greater in patients with AF compared with the control group. Serum galectin-3 levels were also significantly higher in patients with persistent AF than those with paroxysmal AF (median 0.8 ng/ml [interquartile range 0.4 to 1.4] vs 0.5 ng/ml [interquartile range 0.2 to 0.9], p0.001). Multivariate regression analysis demonstrated that serum galectin-3 (odds ratio 87.53, 95% confidence interval 6.06 to 1,265.03, p = 0.001) and LAVI (odds ratio 1.38, 95% confidence interval 1.19 to 1.60, p0.001) were independent predictors of AF. Only LAVI was independently correlated with serum galectin-3 levels in patients with AF in linear regression analysis. In conclusion, serum galectin-3 is significantly elevated and is also significantly correlated with LAVI in patients with AF with preserved left ventricular function.

Aims Atrial fibrosis has been found to be associated with recurrent atrial fibrillation (AF) following catheter ablation. Autoantibodies against M2-muscarinic receptors (anti-M2-R) may play a role in the development of AF by inducing left atrial (LA) fibrosis. In this study, we aim to compare anti-M2-R levels between paroxysmal lone AF patients and healthy control subjects and to investigate the relationship between pre-ablation anti-M2-R level, LA fibrosis quantified by delayed enhancement magnetic resonance imaging (DE-MRI), and AF recurrence following cryoablation. Methods and results Thirty-one patients with paroxysmal lone AF (53.4 ± 8.0 years, 61% male), who underwent cryoballoon-based ablation, along with 31 healthy control subjects were included. Enzyme-linked immunosorbent assay tests to measure serum anti-M2-R levels were performed in both groups and DE-MRI was done to quantify LA fibrosis prior to the ablation in the patients. Anti-M2-R levels were higher in the study population when compared with control subjects [212.4 (103.2–655.5) vs. 73.0 (39.5–299.1) ng/mL, P < 0.001]. Anti-M2-R level predicted moderate-extensive LA fibrosis independent of other measures [odds ratio: 1.26 (95% confidence interval (CI): 1.04–1.53), P = 0.017]. At a mean follow-up of 35.2 ± 3.5 months, nine patients (29.0%) had AF recurrence. In the Cox regression model including pre-ablation anti-M2-R level, LA diameter, LA volume index, and moderate-extensive LA fibrosis, only moderate-extensive LA fibrosis predicted late AF recurrence independent of other measures [hazard ratio: 29.41 (95% CI: 3.52–250.00), P = 0.002]. Conclusion Serum anti-M2-R levels may be associated with the severity of LA fibrosis and may be implicated in the pathophysiology of AF recurrence following cryoablation. Detection of anti-M2-R levels may help select appropriate patients for the procedure.

Background: Recent studies have revealed a relationship between vitamin D deficiency and atherosclerosis. This study aims to investigate the impact of vitamin D deficiency and replacement on markers of subclinical atherosclerosis in young premenopausal women in whom vitamin D deficiency is prevalent. Methods: Thirty-one premenopausal vitamin D deficient women and 27 age and gender-matched control subjects were enrolled in this study. Markers of subclinical atherosclerosis including carotid intima-media thickness (cIMT), flow-mediated dilatation (FMD), endothelial progenitor cell (EPC) count and cytokine levels were determined at baseline. All measurements were repeated at 6-month follow-up in vitamin D-deficient subjects after vitamin D replacement. Results: Vitamin D deficient premenopausal women had lower FMD (9.9 ± 1.3 vs. 13.8 ± 1.7%, p < 0.001) and EPC counts at baseline. This population also had lower IL-10 and higher IL-17 levels. A 6-month vitamin D replacement therapy resulted in a significant increase in FMD (9.9 ± 1.3 vs. 11.4 ± 1.4%, p < 0.001) and EPC counts. Furthermore, cytokine profile shifted toward a more anti-inflammatory phenotype including elevated IL-10 and decreased IL-17 levels. cIMT was not different between patient and control groups and did not change following vitamin D replacement. Change in 25(OH)D and IL-17 levels were independent predictors of the change in FMD measurements following vitamin D replacement. Conclusion: This study demonstrates that endothelial function is impaired in otherwise healthy vitamin D deficient young premenopausal women and improves with 6-month replacement therapy. Immune-modulatory effects of vitamin D may, at least partly, be responsible for its beneficial effects on vascular health.

Interactions with stromal components influence the growth, survival, spread, and colonization capacities of tumor cells. Fibroblasts and macrophages which are responsible for the stroma production and maintenance are of the basic elements found in tumor microenvironment. Cellular density and ratio of stromal cells to tumor cells can also have modulatory effects in cancer. Here, the contribution of fibroblast and/or macrophage cells on the malignant behavior of breast cancer cells was modeled in co-culture systems. Co-cultures were established at different cell densities and ratios with 4T1 breast cancer, NIH/3T3 or 3T3-L1 fibroblast, and J774A.1 monocyte/macrophage cell lines. Flow cytometry-based proliferation, 3D growth on alginate matrix, and matrigel invasion assays were performed to determine the change in the malignant assets of tumor cells. The data were also supported by immunocytochemical and morphological analyses. Co-culturing with fibroblasts (especially, NIH/3T3 cells) significantly supported the proliferation, scattering, and invasiveness of 4T1 cells whereas inclusion of macrophages disrupted this positive influence. On the other hand, the invasion capacity of 4T1 cells was not enhanced in the co-cultures with fibroblasts whose motility were inhibited with pertussis toxin pretreatment. Particularly at low-density seeding in 3D cultures, 4T1 cells could form substantially more spheroids than that of in the co-cultures with fibroblasts. Only, increasing the amount of fibroblasts could restore the 3D-growth. Intriguingly, co-existence of macrophage, fibroblast, and tumor cells in 3D cultures provided a convenient stroma sustaining the spheroid formation and growth. In conclusion, fibroblasts can form a favorable environment for tumor cells' spread and motility whereas restricting their 3D-growth capacity. On the other hand, presence of macrophages may disrupt the influence of fibroblasts and enhance the spheroid formation by the tumor cells.

This study compared the smear layer removing capability and cytotoxicity of NaOCl, EDTA and Oxidative Potential Water (OPW). Fifteen extracted single-rooted human upper incisors were examined in three groups. The root canals were enlarged to the apical foramen with K files to size #60 and irrigated with : (a) NaOCl followed by OPW, (b) OPW during and after instrumentation and (c) NaOCl followed by EDTA and NaOCl. The effect of these irrigants on the smear layer was evaluated using a scanning electron microscope. In vitro cytotoxicity of these irrigants was examined by MTT colorimetric assay. We found that the combination of NaOCl and OPW as well as the application of OPW alone, failed to remove the smear layer from the apical third, whereas the EDTA and NaOCl combination achived complete removal. OPW, when used during and after instrumentation, removed the smear layer in the middle third more effectively than NaOCl followed by OPW. EDTA exerted more cytotoxic effects at all concentrations tested when compared with OPW and NaOC1. In conclusion : (a) OPW was less cytotoxic than other irrigants but did not effectively remove the smear layer, (b) treatment with EDTA followed by NaOC1 efficiently removed of the smear layer, but their cytotoxicity should be considered during endodontic therapy. (J. Oral Sci. 43, 233-238, 2001)

[reaction: see text] A series of water-soluble green perylenediimide (PDI) dyes have been synthesized. On red light excitation, these dyes were shown to be efficient generators of singlet oxygen, and in cell culture media, they were shown to display significant light-induced cytotoxic effects on the human erythroleukemia cell line (K-562). It appears that highly versatile PDI dyes are likely to find new applications in photodynamic therapy.

The inhibitors of cyclooxygenase (COX)-2 play an important role in cancer chemoprevention. Certain COX-2 inhibitors exert antiproliferative and pro-apoptotic effects on cancer cells.In this study, meloxicam, which is an enolic acid-type preferential COX-2 inhibitor, was encapsulated in poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles (NPs) to maintain local high concentration, and its efficacy was determined.NPs were prepared by using salting-out and emulsion-evaporation steps. Meloxicam-loaded NP formulations were evaluated with respect to the drug loading, particle size, polydispersity index, zeta potential, drug release rate, and residual poly(vinyl alcohol) (PVA) percentage. The effects of PLGA and PVA molecular weight variations on the physicochemical properties of NPs were investigated. Stability of meloxicam in NPs was assessed over 3 months. COX-2 expressing human colon adenocarcinoma cell line HT-29 was used in cellular uptake and viability assays.NPs had a spherical shape and a negative zeta potential, and their size ranged between 170-231 nm with a lower polydispersity index. NPs prepared with high molecular weight PLGA were shown to be physically stable over three months at 4°C. The increase in molecular weight of the polymer and emulsifier reduced the in vitro release rate of meloxicam from NPs. Meloxicam-loaded NPs showed cytotoxic effects on HT-29 cells markedly at 800 µM. Cancer cells had high uptake of coumarin-6-loaded NPs.The PLGA NPs developed in this study can be a potentially effective drug delivery system of meloxicam for the treatment of colon cancer.

The plasma levels of most ccmflement compo- nents are maintained by hepatic synthesis (Alper et al., 1969; Cohen, 1976). The nepatocyte is the cell responsible. as shown by studies on the primary cultures of rat and mouse hepatocytes and by studying synthesis of complement com- ponents by hepatocellular carcinoma cell lines such as Hep-3b and Hcp-G2 (Morris et al., 1982a). The most notable exceptions to this rule are Clq, Factor D and properdin. Studies on the acute phase response have shown that comple- ment proteins are acute phase proteins, with increased hcpatocyte synthesis occurring in re- sponse to cytokines such as interleukin 1 (IL-l), interleukin 6 (IL-6), tumour necrosis factor-a and interferon-y (Miura et al., 1987; Andus et al., 1988; Rarnadori et al., 1988; Anthony et al., 1989). Synthesis of complement proteins also occurs in edrahepatic sites. To date it has been shown that all nwima! tissues are able to synthesize at least one complement component [Ruddy and Correspondence to: K. Whdey, Department of Immunology, Leicester Royal Infirmary. Leicester. LEI 5WW. UK Cohen, 1974). In addition, synthesis of comple- ment components occurs in chronically inflamed tissues such as the rheumatoid joint (Moffat et al., 1989). A large number of differem cell types have been shown to synthesize complement com- ponents (Table I). These include mononuclear phagocytes, fibroblasts, endothehal ceils, eplthe- lial cells, alveolar type 2 epithelial cells and adipocytes (Colten, 1976; Morris et al., 1982a; Cook et al., 1987; Cole and Coltzn, 19ir8).

The effects of meconium-stained amniotic fluid (MSAF) on cord blood vascular endothelial growth factor (VEGF) levels have not been explored. The aim of this study was to verify whether MSAF influences cord blood VEGF levels in healthy term neonates and we can use cord blood VEGF levels in infants with MSAF as an indicator of fetal distress.Using an enzyme-linked immunosorbent assay double sandwich method, plasma VEGF levels were determined in 18 healthy term neonates with MSAF and in 16 healthy term neonates without MSAF.VEGF plasma levels were not significantly different between healthy term neonates with or without MSAF.Intrauterine meconium passage could not affect VEGF levels on cord blood in term newborn infants and VEGF level may not be used as an indicator of fetal distress in infants with MSAF.

The aim of this study was to comparatively investigate the effects of 5-azacytidine-C (5-Aza), trichostatin-A (TSA), and all-trans retinoic acid (ATRA) on the mRNA expressions of the sodium and iodine (Na/I) symporter (NIS), thyroglobulin (Tg), thyroid peroxidase (TPO), and the thyroid-stimulating hormone receptor (TSH-R), as well as radioiodine (RAI) uptake in cancer (B-CPAP) and normal (Nthy-ori 3-1) thyroid cell lines. Cell lines were treated with 10 ng/mL of TSA, 5 μM of 5-AZA, and 1 μM of ATRA, according to the MTT (methyl-thiazol-tetrazolium) test results. Additionally, recombinant thyroid-stimulating hormone (rTSH) was also applied, with a selected dose of 100 ng/mL. Following the treatment, NIS, Tg, TPO, and TSH-R mRNA levels were detected by real-time-polymerase chain reaction (RT-PCR) and RAI uptakes were measured by using a well counter as counts/cell number. 5-Aza increased TSH-R mRNA expression in both of the cell lines and decreased TPO, NIS, and Tg mRNA levels in the cancer cell line. In the normal thyroid cell line, 5-AZA increased TPO mRNA levels by 2-fold and made no differences in NIS and Tg mRNA levels. TSA treatment repressed NIS and Tg mRNA levels and made no change on other thyroid-specific genes that were investigated in the cancer cell line. In the normal thyroid cell line, TSA increased TSH-R mRNA levels in 72 hours and created no important difference in the other genes. ATRA repressed the TSH-R mRNA levels in the normal thyroid cell line and increased the TPO and Tg mRNA levels slightly in both the cell lines. Furthermore, in short-term treatment, ATRA repressed the NIS gene expression slightly, but in the long term, this repression turned to basal levels. 5-Aza, TSA, and ATRA did not make any changes in RAI uptake in the cancer cell line, but rTSH increased RAI uptake significantly. In the normal thyroid cell line, TSA and ATRA decreased RAI uptake (to 1/10 and 1/2, respectively), but 5-Aza and rTSH increased RAI uptake significantly (2- and 4-fold, respectively). In our study, we showed an increase in TSH-R gene expression and radioiodine uptake with 5-Aza. Further in vitro and in vivo studies are needed to support our findings and the potential clinical use of this agent. © Mary Ann Liebert, Inc.

There is emerging evidence that matrix metalloproteinases (MMPs) might be involved in blood brain barrier (BBB) breakdown in multiple sclerosis. A group of natural tissue inhibitors of metalloproteinases (TIMPS) regulates proteolytic activity to prevent tissue damage. TIMP-1 and MMP-9 are known to be secreted as heterodimers and TIMP-1 preferentially functions to inhibit MMP-9 activity. In this present study, the effects of IFNbeta-1a on serum MMP-9 and TIMP-1 were evaluated longitudinally during a one-year period. The MMP-9 levels showed no significant changes while TIMP-1 levels gradually and significantly increased during 3rd and 6th months of therapy compared with pretreatment levels.

The widespread use of chemotherapy in the treatment of cancer has led to anxiety about the possible hazards to staff involved in the preparation and administration of cytotoxic agents. Careless handling of antineoplastic drugs may lead to exposure in detectable amounts by means of chemical or biological methods in the body fluids or cell samples but the information about the mutagenic effects of these agents on nurses is limited and inconsistent. DNA damage in peripheral lymphocytes of 30 professional nurses employed in the oncology departments for at least 6 months were examined by the alkaline single cell gel electrophoresis, 'COMET' technique. The results were compared to that of 30 controls with comparable age, sex and smoking habits, not practising in the chemotherapy services. Work characteristics of the exposed nurses and the use of personal protective equipment were also investigated. The DNA damage observed in the lymphocytes of the nurses was significantly higher than the controls (p0.001). The observed DNA damage was found to be significantly lower (p0.001) in nurses applying the necessary individual safety protections during their work. Cigarette smoking was not related to increases in DNA damage, also a significant association was not found between the duration of occupational exposure to antineoplastic drugs and the DNA damage.

We have studied synthesis of the complement components and regulatory proteins of the alternative pathway and the membrane attack complex in synovial membrane. RNA was extracted from synovial tissue of patients with rheumatoid arthritis (RA) or osteoarthritis (OA) as well as from normal synovial membrane. Dot blot analysis showed the presence of mRNAs for all the complement components and regulatory proteins (C3, factor B, factor D, C5, C6, C7, C9, factor H, factor I, S-protein, SP-40, 40, DAF, MCP, CR1, CD59), except for properdin, C8α, C8β and C8γ in all three types of synovial membrane studied. In an attempt to determine which components were synthesised by each cell type, monocytes (mononuclear phagocytes), human umbilical vein endothelial cells (HUVEC), synovial membrane fibroblasts (from normal, OA and RA synovial membrane) and peripheral blood lymphocytes were cultured in vitro and secretion rates of individual components were measured and total cellular RNA analysed by northern blotting. Monocytes secreted properdin, C3, and factor H but not factor B, factor I, C5, C6, C7, C8 or C9. Fibroblasts and endothelial cells secreted factor B, factor H and factor I, but not properdin, C5, C6, C7, C8 or C9. Lymphocytes did not secrete any of these components. mRNAs encoding C3, factor B, factor H, S-protein, SP-40, 40, MCP and DAF were detected in all three other cell types (monocytes, fibroblasts and HUVEC), but factor I and CD59 mRNAs were not detected in monocytes. C5, C6, C7, C8α, C8β, CD8γ and C9 mRNAs were not detected in any of the cell types studied. Cell-specific differences were observed in the expression of the different mRNA species for DAF, MCP and CD59. The results of the present study demonstrate that synthesis of many complement components occurs in normal, RA and OA synovial membrane, and that this may be explained in part by synthesis in mononuclear phagocytes, endothelial cells and fibroblasts. The cellular sources of C5, C6, C7 and C9 mRNAs in synovial membrane have not been determined. The data also show that there are important cell-specific differences in the expression of the genes encoding both the alternative complement pathway components and the membrane regulatory components. These differences require further investigation.

Background The hereditary hemochromatosis gene HFE plays a pivotal role in iron homeostasis. The association between cancer and HFE hetero- or homozygosity has previously been shown including hepatocellular and nonhepatocellular malignancies. This study was performed to compare frequencies of HFE C282Y and H63D variants in Turkish women with breast cancer and healthy controls. Methods Archived DNA samples of Hacettepe University Oncology Institute were used in this study. The HFE gene was investigated by PCR-RFLP. Results All subjects studied were free from C282Y mutation. Thirty-nine patients had H63D mutation and were all heterozygous. H63D allele frequency was 22.2% (39/176) in the breast cancer patients, and 14% (28/200) in the healthy volunteers. Statistical analysis of cases with HFE H63D phenotype showed significant difference between breast cancer and healthy volunteers (P = 0.02). Conclusion Our results suggest that HFE H63D mutation frequencies were increased in the breast cancer patients in comparison to those in the general population. Also, odds ratios (odds ratio = 2.05) computed in this study suggest that H63D has a positive association with breast cancer.