Differentiation of neural stem cells from human olfactory mucosa into dopaminergic neuron-like cells.

The aim of this study was to develop an alternative treatment method for neurodegenerative diseases with dopaminergic neuron loss such as Parkinson's disease by differentiating cells obtained from human olfactory mucosa-derived neural stem cells (hOM-NSCs) with neurotrophic agents in vitro. hOM-NSCs were isolated and subjected to immunophenotypic and MTT analyses. These hOM-NSCs were then cultured in a 3D environment to form neurospheres. The neurospheres were subjected to immunophenotypic analysis and neuronal differentiation assays. Furthermore, hOM-NSCs were differentiated into dopaminergic neuron-like cells in vitro. After differentiation, the dopaminergic neuron-like cells were subjected to immunophenotypic (TH, MAP2) and genotypic (DAT, PITX3, NURR1, TH) characterization. Flow cytometric analysis showed that NSCs were positive for cell surface markers (CD56, CD133). Immunofluorescence analysis showed that NSCs were positive for markers with neuronal and glial cell characteristics (SOX2, NESTIN, TUBB3, GFAP and NG2). Immunofluorescence analysis after differentiation of hOM-NSCs into dopaminergic neuron-like cells in vitro showed that they were positive for a protein specific for dopaminergic neurons (TH). qRT-PCR analysis showed that the expression of dopaminergic neuron-specific genes (DAT, TH, PITX3, NURR1) was significantly increased. It was concluded that hOM-NSCs may be a source of neural stem cells that can be used for cell replacement therapies in neurodegenerative diseases such as Parkinson's disease, are resistant to cell culture, can differentiate into neuronal and glial lineage, are easy to obtain and are cost effective.

Effect of chronic alcohol consumption on myocardial apoptosis in the rat model of isoproterenol-induced myocardial injury and investigation on the cardioprotective role of calpain inhibitor 1.

We investigated the presence of myocardial apoptosis on isoproterenol (ISO)-induced myocardial injury (MI) after long-term high dose alcohol consumption and examined the antiapoptotic role of calpain inhibitor 1. Male Wistar Albino rats (n = 108) were divided into six groups: Control, alcohol (ethanol was given during 30 days for chronic alcohol consumption), MI (150 mg/kg ISO injection at last two days of alcohol consumption), alcohol + MI, alcohol + MI + calpain inhibitor 1 (10 mg/kg inhibitor was injected at 15 min before ISO injections) and Dimethyl Sulfoxide (DMSO) groups. Biochemical, histological, and morphometric methods determined apoptosis levels in the heart tissue of rats. Cytochrome c, caspase 3, and calpain levels were significantly high in alcohol, MI, and alcohol + MI groups. In contrast, mitochondrial cardiolipin content was found to be low in alcohol, MI, and alcohol + MI groups. These parameters were close to the control group in the therapy group. Histological and morphometric data have supported biochemical results. As a result of our biochemical data, myocardial apoptosis was seen in the alcohol, MI, and especially alcohol after MI groups. Calpain inhibitor 1 reduced apoptotic cell death and prevented myocardial tissue injury in these groups. The efficiency of calpain inhibitor was very marked in MI after long-term high dose alcohol consumption.

Erythropoietin Protects the Kidney by Regulating the Effect of TNF-α in L-NAME-Induced Hypertensive Rats.

BACKGROUND/AIMS: Hypertension is the leading cause of death worldwide. Chronic high blood pressure induces inflammation. Tumor necrosis factor (TNF)-α plays a major role in inflammation and also depresses the synthesis of erythropoietin, which exerts protective effects on tissue; however, the mechanism is still unclear. We investigated the protective effect of erythropoietin against tissue damage caused by hypertension in the kidney and whether this effect was suppressed by TNF-α. METHODS: First, we detected the optimum chronic dose for darbepoetin-α (Depo), which is a long-acting erythropoietin analog for rats. We separated 60 female adult rats into 6 groups: control, Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME), L-NAME+Depo, L-NAME+Remicade (an anti-TNF-α antibody), L-NAME+Depo+Remicade, Depo, and control. After 1 month of treatment, we measured cardiovascular parameters, took blood samples, sacrificed the rats, and removed kidneys for analyses. RESULTS: The apoptotic index and the plasma and kidney mRNA levels of TNF-α increased in the L-NAME group and decreased in all other treatment groups. Macrophage accumulation increased in the L-NAME and L-NAME+Remicade groups, while it decreased in the Depo group. The mRNA abundance of TNF receptor 1 (TNFR1) decreased slightly in the Depo group and TNFR2 increased significantly in the same group. CONCLUSION: Erythropoietin protects kidney tissue against hypertension by preventing the apoptotic effects of TNF-α by blocking macrophage accumulation, decreasing TNF-α levels, and switching the TNF-α receptors from the apoptotic receptor TNFR1 to the proliferative receptor TNFR2.

Prenatal alcohol-induced neuroapoptosis in rat brain cerebral cortex: protective effect of folic acid and betaine.

PURPOSE: Alcohol consumption in pregnancy may cause fetal alcohol syndrome (FAS) in the infant. This study aims to investigate prenatal alcohol exposure related neuroapoptosis on the cerebral cortex tissues of newborn rats and possible neuroprotective effects of betaine, folic acid, and combined therapy. METHODS: Pregnant rats were divided into five experimental groups: control, ethanol, ethanol + betaine, ethanol + folic acid, and ethanol + betaine + folic acid combined therapy groups. We measured cytochrome c release, caspase-3, calpain and cathepsin B and L. enzyme activities. In order to observe apoptotic cells in the early stages, TUNEL method was chosen together with histologic methods such as assessing the diameters of the apoptotic cells, their distribution in unit volume and volume proportion of cortical intact neuron nuclei. RESULTS: Calpain, caspase-3 activities, and cytochrome c levels were significantly increased in alcohol group while cathepsin B and L. activities were also found to be elevated albeit not statistically significant. These increases were significantly reversed by folic acid and betaine + folic acid treatments. While ethanol increased the number of apoptotic cells, this increase was prevented in ethanol + betaine and ethanol + betaine + folic acid groups. Morphometric examination showed that the mean diameter of apoptotic cells was increased with ethanol administration while this increase was reduced by betaine and betaine + folic acid treatments. CONCLUSION: We observed that ethanol is capable of triggering apoptotic cell death in the newborn rat brains. Furthermore, folic acid, betaine, and combined therapy of these supplements may reduce neuroapoptosis related to prenatal alcohol consumption, and might be effective on preventing fetal alcohol syndrome in infants.

NT-proBNP predicts impaired myocardial function in newly diagnosed hypertensive patients with preserved ejection fraction.

N-terminal pro-brain natriuretic peptide (NT-proBNP) is an excellent biomarker to diagnose left ventricular (LV) dysfunction. LV myocardial performance index (MPI-Tei index) is commonly used as a measure of combined systolic and diastolic function. We aimed to investigate the relationship between NT-proBNP and tissue Doppler derived MPI in newly diagnosed hypertensive patients with preserved LV ejection fraction (LVEF). We studied 236 patients with newly diagnosed HT (mean age; 52.9 ± 5.2 years). Echocardiographic examination was performed in all patients. LV mass index (LVMI) was calculated. Conventional Doppler indices (E and A waves) were recorded. The MPI value was obtained from the tissue Doppler derived ejection time, isovolumic contraction and relaxation times. The patients were divided into two groups according to the median NT-proBNP value (NT-proBNPlow group <114 pg/ml and NT-proBNPhigh group ≥114 pg/ml). Patients with NT-proBNPhigh were older and had higher levels of glucose and creatinine, lower E/A ratio and higher LVMI and MPI values than patients with NT-proBNPlow. However, LVEF were similar among the groups. Multiple linear regression analysis showed that NT-proBNP was independently associated with age, LVMI, MPI and E/A ratio. Increased NT-proBNP level was independently associated with impaired myocardial performance index in newly diagnosed hypertensive patients with preserved LVEF.

The analysis of boric acid effect on epithelial-mesenchymal transition of CD133 + CD117 + lung cancer stem cells.

Targeting lung cancer stem cells (LC-SCs) for metastasis may be an effective strategy against lung cancer. This study is the first on epithelial-mesenchymal transition (EMT) properties of boric acid (BA) in LC-SCs. LC-SCs were isolated using the magnetic cell sorting (MACS) method. Tumor-sphere formation and flow cytometry confirmed CSC phenotype. The cytotoxic effect of BA was measured by MTT analysis, and the effect of BA on EMT was examined by migration analysis. The expression levels of ZEB1, SNAIL1, ITGA5, CDH1, ITGB1, VIM, COL1A1, and LAMA5 genes were analyzed by RT-qPCR. E-cadherin, Collagen-1, MMP-3, and Vimentin expressions were analyzed immunohistochemically. Boric acid slightly reduced the migration of cancer cells. Increased expression of transcription factor SNAIL (p < 0.001), but not ZEB1, was observed in LC-SCs. mRNA expression levels of ITGB1 (p < 0.01), ITGA5 (p < 0.001), COL1A1 (p < 0.001), and LAMA5 (p < 0.001) increased; CDH1 and VIM decreased in LC-SCs. Moreover, while E-cadherin (p < 0.001) and Collagen-1 (p < 0.01) immunoreactivities significantly increased, MMP-3 (p < 0.001) and Vimentin (p < 0.01) immunoreactivities decreased in BA-treated LC-SCs. To conclude, the current study provided insights into the efficacy and effects of BA against LC-SCs regarding proliferation, EMT, and cell death for future studies.

Relationship between myocardial performance index and severity of coronary artery disease assessed with SYNTAX score in stable coronary artery disease.

BACKGROUND: The aim of the study was to investigate the relationship between severity of coronary artery disease (CAD) assessed with SYNTAX score (SS) and myocardial performance index (MPI) in stable CAD. METHODS: The study participants consisted of 106 consecutive patients (mean age: 57.6 ± 9.5 years) with angiographically proven obstructive stable CAD and 30 patients with nonobstructive CAD (control group) (mean age: 58.0 ± 7.6 years). The MPI was calculated by using pulsed-wave Doppler before coronary angiography. Coronary angiography was performed in all patients. The SS was prospectively calculated in 106 obstructive CAD patients. Patients were partitioned into 3 groups based on SS. Frequencies of risk factors, biochemical and hematological data were recorded in all patients. RESULTS: The SS tertiles were defined as SS(low) ≤ 8 (n = 35), SS(mid) ≤ 17 (n = 36), and SS(high) > 17 (n = 35). The patients with SS(high) group (mean MPI; 0,48 ± 0.06) based on SS had significantly higher MPI values compared with the SS(mid) (mean MPI; 0,44 ± 0.05), SS(low) (mean MPI; 0.43 ± 0.06), and control (mean MPI; 0.41 ± 0.05) groups (P < 0.05 for all). The MPI levels of control group were also lower than compared with SS(mid) group (P = 0.006). The MPI value was significantly correlated with SS (r = 0.564, P < 0.001), diabetes (r = 0.355, P < 0.001), hypertension (r = 0.326, P < 0.001), and ejection fraction (EF) (r = -0.224, P = 0.018) in bivariate analysis. Multivariate regression analysis showed that MPI was independently associated with SS (ß = 0.486, P < 0.001) and diabetes (ß = 0.205, P = 0.028). CONCLUSION: Although the normal EF, MPI value was impaired in proportion to the severity of CAD in patients with stable CAD.

Serum gamma-glutamyltransferase and the burden of atherosclerosis in patients with acute coronary syndrome.

OBJECTIVES: We evaluated the relationship between serum gamma-glutamyltransferase (GGT) levels and the burden of atherosclerosis in patients with acute coronary syndrome (ACS). STUDY DESIGN: This study involved 180 patients (139 male, 41 female; mean age 63±11 years) with the diagnosis of ACS (non-ST elevation myocardial infarction and unstable angina) who underwent coronary angiography on the first day after hospital admission. The burden of atherosclerosis was assessed by the number of involved vessels, and the Gensini and Syntax scores. Serum GGT levels were measured by enzymatic caloric test. RESULTS: Patients with high Syntax scores (>=33) were more frequently diabetic, hypertensive, and had higher GGT and creatinine levels compared to the patients with low Syntax scores (<=23). Similarly, patients with >=3 diseased vessels were more frequently diabetic, hypertensive, and smokers. In addition, these patients were older and had higher serum glucose, urea and GGT levels. Correlation analysis revealed that the level of GGT was significantly associated with Gensini and Syntax scores, number of diseased vessels, and the number of critical lesions (r=0.378 p<0.001, r=0.301 p<0.001, r=0.159 p=0.036, r=0.355 p<0.001, respectively). Multivariate linear regression analysis demonstrated that increased GGT level was an independent risk factor for high Gensini and Syntax scores (p=0.029 and p=0.035, respectively), together with age (p=0.001 and p=0.002, respectively) and serum glucose levels (p=0.017 and p=0.012, respectively). CONCLUSION: Serum GGT levels on admission are associated with increased burden of atherosclerosis in patients with ACS. This may account for the cardiovascular outcomes associated with increased GGT levels.

Increased red cell distribution width level is associated with absence of coronary collateral vessels in patients with acute coronary syndromes.

OBJECTIVES: Several studies have evaluated a relationship between increased red cell distribution width (RDW) and morbidity and mortality of acute coronary syndrome (ACS). In this study, we aimed to investigate the association of serum RDW levels and development of coronary collateral vessel (CCV) in patients with ACS. STUDY DESIGN: We evaluated 226 patients with ACS in this prospective and cross-sectional study. Traditional laboratory and clinical parameters and serum RDW levels were measured on admission. All patients underwent coronary angiography on the first day after admission and patients with >80% stenosis were included in the study. The CCV was graded according to the Rentrop scoring system, and a Rentrop grade 0 was accepted as no CCV development (Group 1), while Rentrop grades 1-2-3 were accepted as presence of CCV development (Group 2). RESULTS: Only levels of RDW were significantly higher in Group 1 than in Group 2 (Group 1 RDW 14.6±1.9, Group 2 RDW 14.1±1.4, p=0.02). The predictive value of serum RDW level for absence of collaterals (sensitivity of 58% and specificity of 54%, area under the receiver operating characteristic (ROC) curve = 0.573) was 13.90. CONCLUSION: We found that high levels of RDW were associated with absence of CCV in patients with ACS.

Stem Cell-Based or Cell-Free Gene Therapy in Chondrocyte Regeneration: Synovial Fluid-Derived Mesenchymal Stem Cell Exosomes.

BACKGROUND: Cartilage injuries are currently the most prevalent joint disease. Previous studies have emphasized the use of stem cells as the effective treatment for regenerating cartilage damage. OBJECTIVE: In this study, considering the difficulties of the cellular therapy method, it was hypothesized that human synovial fluid-derived mesenchymal stem cell (hSFMSC) exosomes as a SC source could be used to treat these injuries as a safer and cell-free therapeutic alternative procedure due to its direct relevance to cartilage regeneration. Moreover, this study aimed to determine the miRNA and target genes required for the formation of SC treatment combined with gene therapy in order to reveal the mechanism of cartilage regeneration and increase its effectiveness. METHODS: MSCs were characterized by flow cytometry, and immunocytochemical and differentiation analyses were done. To characterize functionally isolated exosomes, in vitro uptake analysis was performed. RT-qPCR was used to examine in terms of the advantages of cellular and cell-free therapy, mature human chondroblasts derived by differentiation from hSF-MSCs and human chondrocyte profiles were compared in order to demonstrate the above profile of hSF-MSCs and exosomes isolated from them, and the effectiveness of SC therapy in repairing cartilage damage. RESULTS: According to our findings, the expression level of hsa-miR-155-5p was found to be considerably higher in chondrocytes differentiated from human synovial fluid MSCs than in mature human chondrocytes. These findings were also supported by the TGF-signalling pathway and chondrogenesis marker genes. CONCLUSION: It was concluded that hSF-MSCs and exosomes can be used in the treatment of cartilage damage, and hsa-miR-155-5p can be used as a target miRNA in a new gene therapy approach because it increases the therapeutic effect on cartilage damage.

Developing a Novel Platelet-Rich Plasma-Laden Bioadhesive Hydrogel Contact Lens for the Treatment of Ocular Surface Chemical Injuries.

Permanent injury to corneal limbal stem cells after ocular surface chemical and thermal injuries is a major cause of corneal blindness. In this study, a PRP-laden GelMA hydrogel contact lens is manufactured which is aimed to support the limbal niche after ocular surface insults thereby preventing limbal stem cell failure. GelMA with varying platelet-rich plasma (PRP) concentrations (5%, 10%, and 20%) is photopolymerized using a visible light crosslinking system followed by characterizations of mechanical properties, growth factor release, enzymatic degradation, and in vitro cytotoxicity. The addition of 10% PRP into 10% GelMA hydrogel precursor solution results in the highest tensile and compressive modulus (38 and 110 kPa, respectively) and burst pressure (251±37.66 mmHg). Degradation time varies according to the concentration of the collagenase enzyme tested (0, 2.5, 5, and 40 µg/mL) and is most prolonged with 20% PRP. EGF and TGF-ß release profiles suggest an initial burst release followed by sustained release, most consistent in the 10% PRP sample. Although cell viability decreases on day 1, rapid recovery is observed and is approximately 120% after day 21. PRP-laden GelMA in the form of a contact lens may be a promising biomaterial-based treatment approach for the maintenance of limbal epithelial stem cells after ocular surface insults.

Comparison of exosomes secreted by synovial fluid-derived mesenchymal stem cells and adipose tissue-derived mesenchymal stem cells in culture for microRNA-127-5p expression during chondrogenesis.

This study aimed to investigate the differences between the exosomal microRNA-127-5p expression profiles of human adipose tissue-derived mesenchymal stem cells (hAT-MSCs) and human synovial fluid-derived mesenchymal stem cells (hSF-MSCs) during chondrogenesis in terms of regenerative treatment of cartilage. Synovial fluid-derived mesenchymal stem cells, adipose tissue-derived mesenchymal stem cells, and human fetal chondroblast cells (hfCCs) were directed to chondrogenic differentiation. Alcian Blue and Safranin O stainings were performed to detect chondrogenic differentiation histochemically. Exosomes derived from chondrogenic differentiated cells and their exosomes were isolated and characterized. microRNA-127-5p expressions were measured by Quantitative reverse transcription PCR (qRT-PCR). Significantly higher levels of microRNA-127-5p expression in differentiated hAT-MSCs exosomes, similar to human fetal chondroblast cells, which are the control group in the chondrogenic differentiation process, were observed. hAT-MSCs are better sources of microRNA-127-5p than hSF-MSCs for stimulating chondrogenesis or in the regenerative therapy of cartilage-related pathologies. hAT-MSCs exosomes are rich sources of microRNA-127-5p and can be an essential candidate for cartilage regeneration treatments.

Anti-apoptotic and antioxidant effect of leptin on CCl4-induced acute liver injury in rats.

The aim of this study is to investigate the effect of leptin in rats on carbon tetrachloride (CCl(4)) induced acute liver damage using immunohistochemical methods for apoptosis and biochemical parameters. In this experimental study, 18 Spraque-Dawley rats were divided into three groups viz; control, CCl(4) and CCl(4)+leptin treatment. 0.8 ml/kg olive oil was administered intraperitoneally (i.p.) to the control group and 0.8 ml/kg CCl(4) (1:1 dissolved in olive oil) was administered i.p. to the CCl(4) and CCl(4)+leptin treatment groups, respectively. After 6 h of administrating CCl(4), CCl(4)+leptin treatment group was given i.p. leptin (10 µg/kg). Twenty-four hours after administrating CCl(4) all of the groups were euthanized. Biochemical assessments were performed using serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), plasma tumor necrosis factor alpha (TNF-α) levels and tissue malondialdehyde (MDA), and TNF-α levels. Histological assessments were then performed using Hematoxylin&Eosin (H&E) staining in light microscope and apoptosis assessment using Terminal Transferase dUTP Nick End Labeling (TUNEL)-staining. Serum AST, ALT, ALP and plasma TNF-α levels, tissue MDA and TNF-α levels had all increased in CCl(4) group, but were found to be significantly decreased in CCl(4)+leptin treatment group. Moreover, TUNEL-positive cell counts in liver had significantly increased in CCl(4) group, but decreased in CCl(4)+leptin treatment group (P < 0.05). The results of our study the biochemical, histological and TUNEL-staining showed that leptin has treatment effects on liver CCl(4) induced injury. It plays a role as a potent free radical scavenger, a powerful antioxidant and it also has anti-apoptotic effects.

Changes in carotid intima-media thickness over two years in patients on haemodialysis.

OBJECTIVE: To evaluate the progression of carotid intima-media thickness (CIMT) and to search for possible associations between these changes and other risk factors of atherosclerosis for 2 years in stable patients with chronic renal failure (CRF) on haemodialysis (HD). METHODS: Study population consisted of 22 patients with newly diagnosed CRF. All patients underwent B-mode ultrasonography of common carotid artery for estimating CIMT and the presence of plaques before and after the first HD session (mean 24.22 +/- 2.14 months). The differences in CIMT before and after long-term HD treatment were compared. Acute phase proteins, calcium-phosphate balance and lipid profile were assessed and anthropometric parameters were measured. RESULTS: Mean age was 55 +/- 13 years and 10 (45%) of the patients were female. After long-term HD treatment, (mean 24.22 +/- 2.14 months) the mean value for CIMT (0.57 +/- 0.08 mm) was significantly lower than that at baseline (0.68 +/- 0.12 mm) (p = 0.02). Only male gender and smoking were correlated with baseline CIMT. After long-term HD treatment, age, total cholesterol, LDL cholesterol, and triglyceride were related with CIMT. Diabetes and smoking were correlated with CIMT. Presence of plaque before HD only correlated with creatinine level and after long-term HD treatment only correlated with total cholesterol level. CONCLUSION: We found that CIMT was significantly decreased 2 years after starting HD. An association between CIMT and other atherosclerotic risk factors (such as age, cholesterol, triglyceride etc.) could not be determined due to a small sample size.

[The frequency of aspirin resistance by a modified thrombelastography method and its relationship with clinical and laboratory parameters in patients with stable coronary artery disease]. / Kararli koroner arter hastaligi olanlarda modifiye tromboelastogram yöntemi ile aspirin direnci sikligi ve klinik ve laboratuvar bulgularla iliskisi.

OBJECTIVES: Aspirin is the cornerstone of antiplatelet therapy in cardiovascular medicine. However, aspirin resistance has been demonstrated in 0.4% to 83.3% of aspirin-receiving patients. The aim of this study was to investigate the frequency of aspirin resistance using a modified thrombelastography (mTEG) method and related clinical and biochemical parameters in patients with stable coronary artery disease (CAD), who received 100 mg/day aspirin. STUDY DESIGN: The study included 168 patients (115 males, 53 females; mean age 60±8 years) with stable CAD, receiving aspirin at a dose of 100 mg/day. Aspirin responsiveness was determined using mTEG, where aspirin resistance was defined as arachidonic acid-induced whole blood platelet aggregation inhibition (PAI) of less than 50%. RESULTS: Aspirin resistance was detected in 27 patients (16.1%). Platelet aggregation inhibition showed negative correlations with hyperlipidemia, smoking, spironolactone use, systolic blood pressure, pulse pressure, and total cholesterol and fibrinogen levels. In multivariate regression analysis, only fibrinogen level (OR=1.063, p=0.010) and pulse pressure (OR=1.197, p=0.023) were found to be independent indicators of aspirin resistance and PAI. In ROC analysis, cut-off values of 50 mmHg for pulse pressure and 400 mg/dl for fibrinogen level predicted aspirin resistance with 88.9% and 74% sensitivity and 64.4% and 68% specificity, respectively. CONCLUSION: Our findings suggest that measurements of fibrinogen level and pulse pressure may be used as easy and reliable methods in predicting aspirin resistance.

The potential therapeutic effect of klotho on cell viability in human colorectal adenocarcinoma HT-29 cells.

Klotho is an anti-aging, anti-inflammator, and anti-oxidative protein and has been shown to important role in tumorigenesis, proliferation, survival, autophagy, and resistance to tumor suppressor effects in several types of human cancers. In this study, we aimed to investigate possible anti-tümör and apoptotic effects of exogen klotho in human colorectal adenocarcinoma cells (HT-29) and healthy colon cells (CCD 841 CoN). The WST-8 test was used to determine the half-maximum inhibitory concentration (IC50) of the klotho protein. AO-PI fluorescent staining techniques and Annexin V-PI flow cytometry was utilized to observe and detect the apoptosis of cancer cells induced by klotho. Our results demonstrated that klotho had a cytotoxic effect against colorectal adenocarcinoma cells in a dose-dependent manner. Our Annexin V-PI flow cytometric and AO-PI fluorescent analyses showed that klotho induced quantitative and morphological changes that indicate apoptotic induction in the human colorectal adenocarcinoma. This study results proved for the first time that klotho may be an effective potential therapeutic agent that may be used in adjuvant therapy in human colorectal adenocarcinoma it does not affect selectively healthy colon cells and but leading cancer cells to apoptosis.

[Usefulness of high-sensitivity CRP increases during circadian rhythm for prediction of long-term cardiovascular events in patients with stable coronary artery disease]. / Kararli koroner arter hastalarinda günlük döngü içinde artis gösteren yüksek duyarlikli CRP düzeyinin uzun dönem kardiyovasküler olaylari öngörmede kullanilabilirligi.

OBJECTIVES: We investigated the value of circadian variations in high-sensitivity C-reactive protein (hs-CRP) levels in prediction of long-term cardiovascular events (CVE) in patients with stable coronary artery disease (CAD). STUDY DESIGN: The study included 94 patients (70 men, 24 women; mean age 58 ± 9 years) with stable CAD. High-sensitivity CRP levels were measured at six-hour intervals, namely, morning (06:00), midday (12:00), evening (18:00), and midnight (24:00). Absolute change in hs-CRP (absolute ΔCRP) was calculated by subtracting the midday hs-CRP level from that of the morning. Relative change in hs-CRP (relative ΔCRP) was calculated by dividing absolute ΔCRP by the midday hs-CRP level. The patients were followed-up for a mean of 40.2 ± 8.0 months for monitoring of CVE. RESULTS: During the follow-up period, CVE occurred in 24 patients (25.5%). Patients who developed CVE exhibited significantly higher serum creatinine, B-type natriuretic peptide, morning, evening, and midnight hs-CRP levels, absolute and relative ΔCRP, and left atrial end-diastolic diameter compared to patients without CVE (p<0.05). In logistic regression analysis, only left atrial end-diastolic diameter and absolute ΔCRP were independent predictors of CVE (OR=1.11, 95% CI 1.003-1.236, p=0.044 and OR=1.58, 95% CI 1.195-2.090, p=0.001, respectively). Every 1 mg/l increase in absolute ΔCRP represented a 58.1% increase in CVE risk. In receiver operating characteristics curve analysis, the cut-off value of 2 mg/l for absolute ΔCRP predicted CVE with 89.5% sensitivity and 84.2% specificity. CONCLUSION: Our findings suggest that absolute circadian increases in hs-CRP levels may be helpful in predicting long-term CVEs in patients with stable CAD.

Enhancement of Apo2L/TRAIL signaling pathway receptors by the activation of Klotho gene with CRISPR/Cas9 in Caco-2 colon cancer cells.

Human Klotho gene has many known functions such as anti-aging and anti-tumor, and decreased expression of this gene causes malignant formations in most types of cancer, including colon cancer. Interacting with TRAIL death receptors (DR4 and DR5) induces an apoptotic effect in cancer treatments by reducing the proliferation of cancer cells. The present study aimed to investigate downstream effect of overexpression of Klotho gene, which is known to have an antitumor effect on resistant human colon cancer cells, by examining its action on TRAIL death and decoy (DcR1 and DcR2) receptors for the first time. For this purpose, upregulation of human Klotho gene was achieved with CRISPR/Cas9-mediated system in resistant human colon cancer Caco-2 cells. To determine the effect of upregulation of Klotho gene on cancer cells evaluations with flow cytometry, WST-8, qRT-PCR, ELISA, and immunohistochemical analysis were performed. Then, Klotho gene was knocked out and its apoptotic effect was tested to find out whether it is due to overexpression of Klotho gene or not. Our results indicate that overexpression of Klotho gene in Caco-2 cells via CRISPR/Cas9-sensitized TRAIL death receptor DR4 suppresses the proliferation of cells by leading to apoptosis. Thus, this study conducted on apoptosis-resistant colon cancer cells may bring new insights about the role of Klotho gene in colon cancer.

D-dimers are associated with coronary artery disease severity assessed using Syntax and Syntax II scores in patients with ST elevation myocardial infarction.

INTRODUCTION AND OBJECTIVES: D-dimers are a determinant of hypercoagulable state and have been found to be related to acute coronary syndromes. We aimed to establish the association between increased D-dimer levels and coronary artery disease (CAD) severity using SYNTAX Score (SS) II in patients with ST elevation myocardial infarction (STEMI) who underwent primary percutaneous coronary intervention (PCI). METHODS: This retrospective study included 300 consecutive patients (81.7% males, mean age 55±12 years) with STEMI who underwent a primary PCI. Patients were divided into two groups according to their median SSII [SSII<25 as a low group (n=151) and SSII≥25 as a high group (n=149)]. Blood samples for D-dimers and the other biochemical parameters were obtained from each patient at admission. RESULTS: When compared with the low SSII group, frequency of female gender, no-reflow phenomenon, D-dimer levels, thrombus score, creatine kinase MB and troponin were significantly higher, whereas left ventricular ejection fraction (LVEF) and glomerular filtration rate (GFR) were lower in the high SSII group (p<0.05, for all). D-dimer levels, thrombus score, LVEF, GFR and no-reflow phenomenon were independent predictors of CAD severity (p<0.05, for all). Receiver operating characteristic curve analysis showed that the D-dimer cut-off value for predicting the severity of CAD was 0.26 µg/ml (69.8% sensitivity and 65.6% specificity, p<0.001). CONCLUSION: Increased D-dimer levels are associated with the severity of CAD based on Syntax Score II, in patients with STEMI who successfully underwent revascularization with a primary PCI.

The role of prokineticins in recurrent implantation failure.

The aim of the present study was to investigate the expression patterns of prokineticins (PROK) and prokineticin receptors (PROKR) in the endometrium of women with recurrent implantation failure (RIF). Fifteen (15) women with RIF and 15 fertile controls were enrolled in this study. Endometrial samples were taken from study participants with an endometrial biopsy cannula during the implantation window. Real time polymerase chain reaction and immunohistochemistry were used to determine PROK/PROKR mRNA expression and protein localization, respectively. PROK1 mRNA levels were 6.09 times higher compared to endometrial samples obtained from women with RIF than in samples obtained from fertile controls, whereas PROKR1 mRNA levels were 2.46 times lower in endometrial samples obtained from women with RIF than in samples from fertile controls. In addition, decreased PROKR1 was supported by immunohistochemistry analysis at protein level. There was no statistically significant difference between women with RIF and fertile controls regarding PROK2 and PROKR2 levels. Altered expression of the PROK1/PROKR1 system could be one of the numerous abnormalities in the endometrium of women with RIF.