Effects of TGF-β1 Overexpression on Biological Characteristics of Human Dental Pulp-derived Mesenchymal Stromal Cells

OBJECTIVE: The aim of our study was to investigate the effect of Transforming growth factor beta-1 (TGF-β1) gene therapy on the surface markers, multilineage differentiation, viability, apoptosis, cell cycle, DNA damage and senescence of human Dental Pulp-derived Mesenchymal Stromal Cells (hDPSC). METHODS: hDPSCs were isolated from human teeth, and were cultured with 20% Fetal Bovine Serum (FBS) in minimum essential media-alpha (α-MEM). TGF-β1 gene transfer into hDPSCs was performed by electroporation method after the plasmid was prepared. The transfection efficiency was achieved by using western blot and flow cytometry analyses and GFP transfection. Mesenchymal stem cell (MSC) markers, multilineage differentiation, cell proliferation, apoptosis, cell cycle, DNA damage and cellular senescence assays were performed by comparing the transfected and non-transfected cells. Statistical analyses were performed using GraphPad Prism. RESULTS: Strong expression of TGF-β1 in pCMV-TGF-β1-transfected hDPSCs was detected in flow cytometry analysis. TGF-β1 transfection efficiency was measured as 95%. Western blot analysis showed that TGF-β1 protein levels increased at third and sixth days in pCMV-TGF-β1-transfected hDPSCs. The continuous TGF-β1 overexpression in hDPSCs did not influence the immunophenotype and surface marker expression of MSCs. Our results showed that TGF-β1 increased osteogenic and chondrogenic differentiation, but decreased adipogenic differentiation. Overexpression of TGF-β1 increased the proliferation rate and decreased total apoptosis in hDPSCs (p<0.05). The number of cells at “S” phase was higher with TGF-β1 transfection (p<0.05). Cellular senescence decreased in TGF-β1 transfected group (p<0.05). CONCLUSIONS: These results reflect that TGF-β1 has major impact on MSC differentiation. TGF-β1 transfection has positive effect on proliferation, cell cycle, and prevents cellular senescence and apoptosis.

Acetylsalicylic acid resistance in patients with Type 2 diabetes mellitus, prediabetes and non-diabetic coronary artery disease

Several studies have demonstrated the beneficial role of antiplatelet therapy with acetylsalicylic acid [ASA] at atherosclerotic vascular disease. Antiaggregant effect of ASA is not uniform in all patients. Purpose of the present study is to evaluate the prevalence of ASA resistance in patients with type 2 diabetes mellitus [T2DM], pre-diabetes and non-diabetic coronary artery disease [CAD]. Effect of ASA was assessed using the platelet function analyzer [PFA-100] system. Resistance to ASA was defined as a normal collagen/epinephrine induced closure time after one week of ASA therapy. Patients with non-diabetic CAD, pre-diabetes and T2DM were compared. ASA resistance was found in 26 [37.1%], 6 [17.6%] and 41 [26.5%] patients in the groups, respectively [p=0.154]. ASA resistance was found to be significantly higher in men, smokers and insulin users, besides this it was found to be significantly lower in beta blocker [BB] users, angiotensin converting enzyme inhibitor [ACEI] users with univariate analysis. However insulin usage was found to be the single effective parameter on ASA resistance in multivariate analysis. There was no difference with regard to ASA resistance between groups. While ASA resistance was higher in men, smokers and insulin users, it was lower in patients using BBs and ACEIs

The Independent Relationship of Systemic Inflammation With Fragmented QRS Complexes in Patients With Acute Coronary Syndromes

BACKGROUND AND OBJECTIVES: QRS complex fragmentations are frequently seen on routine electrocardiograms with narrow or wide QRS complex. Fragmented QRS complex (fQRS) is associated with increased morbidity and mortality, sudden cardiac death and recurrent cardiovascular events. In this study, we aimed to interrogate the relationship of systemic inflammation with the presence of fQRS in patients with acute coronary syndromes (ACS). SUBJECTS AND METHODS: Two-hundred and twenty eligible patients with ACS that underwent coronary angiography were enrolled consecutively in this study. Patients with significant organic valve disease and those with any QRS morphology that had a QRS duration > or =120 ms as well as patients with permanent pacemakers were excluded from this study. RESULTS: Patients with fQRS were of a higher age (p=0.02), had increased C-reactive protein (CRP) levels (p<0.001), prolonged QRS time (p<0.001), extent of coronary artery disease (CAD) (p<0.001), creatine kinase-MB (CK-MB) levels (p=0.006) and Q wave on admission electrocardiography (p<0.001) in comparison to patients with non-fragmented QRS. When we performed multiple logistic regression analysis, fQRS was found to be related to increased CRP levels {odds ratio (OR): 1.2, 95% confidence interval (CI): 1.045-1.316, p=0.007}, QRS duration (OR: 1.1, 95% CI: 1.033-1.098, p<0.001), extent of CAD (OR: 1.5, 95% CI: 1.023-2.144, p=0.037), Q wave (OR: 2.2, 95% CI: 1.084-4.598, p=0.03) and CK-MB levels (OR: 1.0, 95% CI: 1.001-1.037, p=0.04) independently. CONCLUSION: In our study, we found that fQRS was independently related to increased CRP. Fragmented QRS that may result as an end effect of inflammation at cellular level can represent increased cardiac risk by different causative mechanisms in patients with ACS.

Absence and Resolution of Fragmented QRS Predict Reversible Myocardial Ischemia With Higher Probability of ST Segment Resolution in Patients With ST Segment Elevation Myocardial Infarction

BACKGROUND AND OBJECTIVES: Fragmented QRS complexes (fQRS) are associated with increased morbidity and mortality. The causative relationship between fQRS and cardiac fibrosis has been shown, but whether the presence and the number of fQRS on admission of electrocardiogram (ECG) predicts ST segment resolution in patients undergoing primary percutaneous coronary intervention (p-PCI) has not been investigated until now. SUBJECTS AND METHODS: This study included one hundred and eighty-four consecutive patients with ST elevation myocardial infarction (STEMI) who underwent p-PCI. The presence or absence of fQRS on pre and post-PCI ECG and their relation with myocardial infarction and reperfusion parameters were investigated. RESULTS: Patients with fQRS on admission of ECG or newly developed fQRS after p-PCI had increased inflammatory markers, higher cardiac enzyme levels, increased pain to balloon time, prolonged QRS time, more extended coronary involvement and more frequent Q waves on ECG in comparison to patients with absence or resolved fQRS. The presence and higher number of fQRS on admission or post-PCI ECGs were significantly related with low percent of ST resolution and myocardial reperfusion parameters. The area under the receiver operating characteristics curve values for the presence and number of fQRS to detect Thrombolysis in Myocardial Infarction Blush Grade 0 and 1, were 0.682 and 0.703. CONCLUSION: In our study, fQRS was significantly related to infarction and myocardial reperfusion parameters before and after p-PCI. Successful myocardial reperfusion by p-PCI caused the reduction in number of fQRS and QRS time with higher ST resolution. fQRS may be useful in identifying the patients at higher cardiac risk with increased ischemic jeopardized or infarcted myocardium, and persistent or newly developed fQRS may predict low percent of ST segment resolution in patients undergoing p-PCI.

Clinicopathologic Characteristics and Therapeutic Outcomes of Primary Gastrointestinal Non-Hodgkin's Lymphomas in Central Anatolia, in Turkey

Primary gastrointestinal lymphoma is a common presentation of non-Hodgkin's lymphoma. The main controversy arises when many aspects of its classification and management are under discussion, particularly regarding roles for surgical resection. The aim of this study was to evaluate clinicopathologic characteristics and the therapeutic outcome of primary gastrointestinal non-Hodgkin's lymphoma. We carried out a retrospective analysis of 74 patients who were presented to our center with histopathological diagnosis of primary gastro-intestinal non-Hodgkin's lymphoma between 1990 and 2001. All patients have been staged according to Lugano Staging System. For histopathological classification, International Working Formulation was applied. The treatment choice concerning the surgical or non-surgical management was decided by the initially acting physician. Treatment modalities were compared using the parameters of age, sex, histopathological results, stage, and the site of disease. Of the 74 patients, 31 were female and 43 were male, with a median age of 49 years (range 15-80). The stomach was the most common primary site and was seen in 51 of 74 patients (68.9%). The intermediate and high grade lymphomas constituted 91.9% of the all cases. In a median follow-up of 29 months (range 2-128), 20 out of 74 patients died. There was a three year overall survival rate in 65.4% of all patients. The three year overall survival rate was better in stage I and II1 patients who were treated with surgery plus chemotherapy (+/-RT) than those treated with chemotherapy alone (93.7% vs. 55.6%, p<0.05). The stage and presence of B symptoms affected the disease free survival and overall survival significantly, but the histopathologic grade only affected the overall survival. On the basis of these results, we suggest that surgical resection is necessary before chemotherapy in early stage (stage I and II1) patients with gastrointestinal non-Hodgkin's lymphomas because of the significant survival advantage it would bring to the patient.

Acute Lymphoblastic Leukemia Associated with Brucellosis in Two Patients with Fever and Pancytopenia

Brucellosis is a disease involving the lymphoproliferative system, which may lead to changes in the hematological parameters; however, pancytopenia is a rare finding. However, malignant diseases in association with brucellosis are rarely the cause of pancytopenia. Herein, two cases with fever and pancytopenia, diagnosed as simultaneous acute lymphoblastic leukemia and brucellosis are presented. Anti-leukemic therapy and brucellosis treatment were administered simultaneously, and normal blood parameters obtained. The first patient is in complete remission; the other recovered from the brucellosis, but later died due to a leukemic relapse.

Influence of Sex and Age on the Activity of Antioxidant Enzymes of Polymorphonuclear Leukocytes in Healthy subjects

In this study, the main antioxidant enzymes (AOE) of glutathione peroxidase (GPX), superoxide dismutase (SOD), catalase (CAT) and myeloperoxidase (MPO) were identified, and the influence of sex and age in healthy human polymorphonuclear leukocytes (PMNL) was determined. The SOD, GPX, CAT and MPO activities were investigated in intestinal parasite negative human PMNL from 109 healthy subjects aged from 6 to 70 years (55 males and 54 females) using simple and sensitive enzyme assays. Blood cells, such as eosinophils, platelets, neutrophils, monocytes, and macrophages also synthesize antioxidant enzymes (AOE). They constitute an important proportion and are also the major participants in a number of pathological conditions that suggest the involvement of AOE. A linear effect of age on SOD activity (p < 0.05) both in males and females was found. A similar effect with GPX activity (p < 0.05) was observed in males only. This showed that the activities of all these enzymes increase with age. In addition, SOD activity was significantly higher in females than males between the age of 19 and 70 years (p < 0.001). This analysis also showed that there is a negative correlation between the CAT-GPX (p < 0.05) activities and positive correlations between MPO-GPX (p < 0.05) activities only in females. No correlation among the other enzyme activities was found in either sex group. This study showed the activities of antioxidant enzyme activities and the correlations of these enyzmes activities with each other in healthy human PMNLs were age- and sex-dependent. This information may assisit in understanding the importance of antioxidant enzymes in the physiological and pathological conditions associated with PMNL.