Genotyping of single nucleotide polymorphism by probe-gated silica nanoparticles.

The development of simple, reliable, and rapid approaches for molecular detection of common mutations is important for prevention and early diagnosis of genetic diseases, including Thalessemia. Oligonucleotide-gated mesoporous nanoparticles-based analysis is a new platform for mutation detection that has the advantages of sensitivity, rapidity, accuracy, and convenience. A specific mutation in ß-thalassemia, one of the most prevalent inherited diseases in several countries, was used as model disease in this study. An assay for detection of IVS110 point mutation (A > G reversion) was developed by designing probe-gated mesoporous silica nanoparticles (MCM-41) loaded with reporter fluorescein molecules. The silica nanoparticles were characterized by AFM, TEM and BET analysis for having 180 nm diameter and 2.83 nm pore size regular hexagonal shape. Amine group functionalized nanoparticles were analysed with FTIR technique. Mutated and normal sequence probe oligonucleotides)about 12.7 nmol per mg nanoparticles) were used to entrap reporter fluorescein molecules inside the pores and hybridization with single stranded DNA targets amplified by PCR gave different fluorescent signals for mutated targets. Samples from IVS110 mutated and normal patients resulted in statistically significant differences when the assay procedure were applied.

Detection of viruses by probe-gated silica nanoparticles directly from swab samples.

Viral infection has been one of the major health issues for human life. The real-time reverse transcription polymerase chain reaction (RT-PCR)-based detection has primarily been used for virus detection as a highly reliable procedure. However, it is a relatively long and multi-stage process. In addition, required skilled personnel and complex instrumentation presents difficulties in large scale monitoring efforts. Therefore, we report here a direct and fast detection method for CoV-2 genome as applied in the nose-throat swab samples without any further processing. The detection principle is based on fluorescein-loaded mesoporous silica nanoparticles capped by specific gene sequences probes immobilized on the surface of the nanoparticles. Upon hybridization with the target viral genome, the fluorescein molecules were released from the mesopores. Testing with synthetic oligonucleotides, the NSP12 gene-based detection resulted in a strong signal. Target detection time could be optimized to 15 min and the limit of detection was 1.4 RFU with 84% sensitivity with clinical samples (n = 43).

Ezetimibe therapy and its influence on oxidative stress and fibrinolytic activity.

OBJECTIVE: The effect of ezetimibe on blood lipids, oxidative stress, and fibrinolytic activity in hyperlipidemic patients was investigated after three months of therapy. METHODS: Thirty hyperlipidemic patients were treated for twelve weeks with ezetimibe 10 mg/day. A healthy control group with matching age and gender was also included. Fasting blood glucose, lipid parameters, paraoxonase (PON1), protein carbonyl (PCO), oxidized LDL (oxLDL), 8-isoprostane (ISOPR), total antioxidant capacity (TAC) levels, tissue-type plasminogen activator (tPA), plasminogen activator inhibitor type-1 (PAI-1), and PAI-1/t-PA levels were evaluated. RESULTS: Ezetimibe therapy for twelve weeks led to changes in lipid profile in accordance with the literature. Fibrinolytic activity parameters, PAI-1/tPA and tPA-1 decreased, whereas PAI-1 levels did not change significantly. Antioxidant parameters, serum PON1 activity, and TAC levels increased significantly compared with the basal values. Oxidant parameters, oxLDL, ISOPR, and PCO (which is an indicator of oxidative protein damage) decreased significantly after therapy. CONCLUSIONS: Ezetimibe therapy has beneficial effects on fibrinolytic activity and homeostasis between oxidant and antioxidant activity in hyperlipidemic patients This may be through lowering lipid levels or other mechanisms such as decreasing insulin resistance and the pleiotropic effects of the drug.

[Analysis of the alterations of trace elements in plasma and tissue, in lung cancer]. / Akciger kanserinde serum ve dokuda eser element degisimlerinin incelenmesi.

In our research, we aimed to analyse the relationship between the lung cancer and the zinc (Zn) and the copper (Cu) which are important trace elements for the organisms in cases containing lung cancer. The research included 20 patients who have been operated as a result of primary lung cancer. In the research, the Zn and Cu and Cu/Zn levels in the patient's cancerous and normal lung tissues and serum (pre-operative, post-operative day first and day fifth) were measured (pre-operative, post-operative day first and post-operative day fifth serum albumin levels were also measured). It was found out that the Cu level in the tissue with tumour has been significantly higher than that of the normal tissue (p< 0.01); that the Zn level in the normal tissue has been significantly higher than the tissue with tumour (p< 0.001) and that the Cu/Zn rate in the tissue with tumour has been significantly higher than the normal tissue (p< 0.001). Regarding the serum values, in the post-operative day first, a significant decrease has been viewed in the levels of Cu, Zn and albumin compared to the pre-operative period (p< 0.001); in the post-operative day first serum Cu/Zn rate has significantly increased compared to the pre-operative period (p< 0.001); in the post-operative day fifth serum albumin levels have significantly decreased compared to the pre-operative period (p< 0.001); in the post-operative day fifth, serum Cu/Zn rate has significantly decreased compared to the post-operative day first (p< 0.01). As a result of the research we performed, we believe that the Cu/Zn level is a more significant data regarding the research of the cancer than the levels of Zn and Cu. We also believe that the Cu/Zn level in the patients with lung cancer should not be used as a diagnostic test, but rather as an indicator of the weakening of the antioxidant defence in patients.

Abdominal aortic aneurysm or aortic occlusive disease: role of trace element imbalance.

The nature of the relationship between atheromatous disease and degenerative aneurysm is yet to be defined. The purpose of this study was to compare tissue Fe, Cu, Zn, and thiobarbituric acid reactive substances (TBARS) levels (as a marker of lipid peroxidation) in the abdominal aorta in relation to the development of aneurysmal and occlusive disease in the infrarenal aorta. This was a prospective clinical study in an institutional referral center, in hospitalized patients. Eighty male patients who underwent surgery for abdominal aortic aneurysm (AAA) or aortic occlusive disease (AOD) were included in the study. Age, risk factors and comorbid conditions were recorded, including diabetes mellitus, hypertension, coronary artery disease, smoking, and chronic obstructive pulmonary disease. Aortic wall biopsies were collected at operation from the anterolateral section of the infrarenal aorta. Tissue Fe, Cu, Zn, and TBARS levels were determined. The mean age of the AAA group was 66.2 (56-75) years and of the AOD group 57.8 (47-72) years (p <0.001). There was a higher prevalence of hypertension in AAA patients compared to AOD patients (62.5%, 35% respectively; p <0.05). The comparison of tissue Zn levels showed no significant difference. Tissue levels of Fe, Cu, and TBARS were found to be higher in the AAA group, compared with the AOD group (p<0.001 for each). These results suggest that higher oxidative stress as a result of higher Fe and Cu levels in the AAA, compared with AOD, may be one of the contributing factors in aneurysmal formation as a result of promoted wall erosion.

Association of plasma viscosity with cardiovascular risk factors in obesity: an old marker, a new insight.

OBJECTIVE: Although obesity is related with cardiovascular disease, the exact mechanism of the relationship is not fully understood. We aim to examine the relationship between plasma viscosity and obesity as a cardiovascular disease risk factor in obese and non-obese groups. METHODS: We recruited 75 obese subjects (mean age: 40.2+/-8.4 years, Body Mass Index: 33.61+/-2.57 kg/m(2)) who were admitted to the Clinic of Endocrinology and Metabolism of Cerrahpasa Medical Faculty. As a non-obese group (n=70, mean age: 41.78+/-9.7 years, Body Mass Index: 21.84+/-3.42 kg/m(2)) healthy subjects from medical and laboratory staff were selected. Plasma viscosity and lipid profile were measured and atherogenic index was calculated as atherogenic risk factors. RESULTS: Plasma viscosity, total cholesterol and LDL-cholesterol levels and atherogenic index were significantly increased in obese group compared to non-obese group for each p<0.001. We found no significant difference in plasma fibrinogen, insulin, albumin and HDL-cholesterol levels between obese and non-obese groups. Plasma viscosity was correlated with total cholesterol and atherogenic index only in the obese group (p<0.05 and p<0.05 respectively). In the non-obese group regarding PV, we determined a positive correlation with triglycerides (r: 0.470, p<0.05) and negative correlation with HDL-C (r: -0.518, p<0.05). CONCLUSION: Plasma viscosity, an early atherosclerotic risk factor, might be helpful in the assessment of cardiovascular risk in obese subjects along with classical cardiovascular risk factors such as plasma cholesterol and atherogenic index.

Effects of cigarette smoking on hemorheologic parameters, plasma osmolality and lung function.

BACKGROUND: Cigarette smoking deteriorates human health via vascular disorders, cancer and especially respiratory diseases. The aim of this study is to investigate effects of cigarette smoking on hemorheologic parameters, plasma osmolality and lung function in individuals without diagnosis of chronic obstructive pulmonary disease (COPD). METHODS: Patients diagnosed without COPD utilizing respiratory function test were enrolled in the study with three groups, ex-smokers (n = 21), current-smokers (n = 35) and never-smokers (n = 43). Hemorheologic parameters and plasma osmolality were measured in hemorheology laboratory. SPSS 17.0 was used for statistical analysis. RESULTS: Blood and plasma viscosity, fibrinogen and hematocrit levels, mean corpuscular volume and mean corpuscular hemoglobin concentration were significantly elevated in ex-smokers and current-smokers compared to never-smokers. The standardized red blood cell deformability and oxygen delivery index and lung function were statistically lower in current-smokers than never-smokers. Pulmonary blood flow rate was statistically lower in current-smokers and ex-smokers than never-smokers. Plasma osmolality was statistically significantly higher in ex-smokers and current-smokers than never-smokers. CONCLUSIONS: Our findings clearly show that cigarette smoking has severe effects on hemorheologic parameters, plasma osmolality and lung function even in individuals without COPD. Blood and plasma viscosity with plasma osmolality might be useful markers to detect early hemorheologic-hemodynamic alterations in cigarette smokers.

Plasma homocysteine levels in obese and non-obese subjects with or without hypertension; its relationship with oxidative stress and copper.

OBJECTIVES: The relationship between plasma total Homocysteine (tHcy) and oxidative stress and plasma levels of lipids, insulin and copper levels were investigated in obese and nonobese hypertensives. DESIGN AND METHODS: Plasma tHcy levels were determined by an enzyme immunoassay method. Plasma lipid peroxidation levels were measured as thiobarbituric acid reactive substances (TBARS) by spectrophotometric methods. Plasma levels of copper and insulin were measured by atomic absorption spectrophotometer and electrochemiluminescence method, respectively. RESULTS: Plasma tHcy, copper and insulin levels did not differ in nonobese hypertensives compared to nonobese normotensives. Plasma TBARS levels were significantly increased in nonobese hypertensives when compared to nonobese normotensives (p < 0.001). Plasma tHcy, TBARS, copper and fasting insulin levels were significantly higher in obese normotensives and hypertensives than in nonobese normotensives and hypertensives, respectively (for each comparison; p < 0.001). There was a significant difference in plasma tHcy, TBARS and copper levels between obese subjects with or without hypertension (for each comparison p < 0.01). The univariate analyses demonstrated a significant positive correlation between tHcy and TBARS (coefficient +/- SE, 0.411 +/- 0.115, p < 0.01) and copper (coefficient +/- SE, 0.425 +/- 0.135, p < 0.01) in obese subjects. In a multivariate regression analysis in obese subjects tHcy was positively correlated with TBARS (coefficient +/- SE, 0.480 +/- 0.155, p < 0.01) and copper (coefficient +/- SE, 0.486 +/- 0.140, p < 0.01). CONCLUSIONS: We hypothesize that in the presence of other traditional risk factors, Hcy may have a permissive role in the endothelium damage even within the normal range and this role may be related to free radical generating systems. Therefore, modest elevation of plasma Hcy may causally be involved in the pathogenesis of atherosclerosis and/or cardiovascular disease.

Plasma viscosity as an early cardiovascular risk factor in hirsute women with eumenorrhea or oligomenorrhea.

To investigate plasma viscosity, a hemorheologic variable, in oligomenorrheic and eumenorrheic hirsute women.Descriptive study. Endocrinology and metabolism clinic. Twenty five hirsute women with oligomenorrhea and eumenorrhea and 30 age- and weight-matched healthy, eumenorrheic, nonhirsute women. Plasma viscosity. Plasma viscosity was significantly elevated in hirsute women compared with eumenorrheic nonhirsute women. Significant correlations were observed between plasma viscosity and fasting insulin level, cholesterol level, and diastolic blood pressure. Hirsute women with oligomenorrhea or eumenorrhea had a significantly elevated plasma viscosity compared with healthy controls.

Relationship between plasma leptin and zinc levels and the effect of insulin and oxidative stress on leptin levels in obese diabetic patients.

Leptin is thought to be a lipostatic signal that contributes to body weight regulation. Zinc plays an important role in appetite regulation also. Our aim is to evaluate the relationship between leptin and zinc in obese and nonobese type 2 diabetic patients and its relationship with oxidative stress and insulin. We studied 25 nonobese nondiabetic women (controls); 35 nonobese diabetic women; and 45 obese diabetic women. Plasma leptin concentration was determined by immunoradiometric assay. Thiobarbituric acid reactive substances (TBARS), markers of oxidative stress, were assayed by the spectrofotometric method. Plasma levels of zinc and insulin were measured by atomic absorption spectrophotometer and electrochemiluminescence methods, respectively. We found that nonobese diabetic patients had significantly lower zinc and higher TBARS levels than control subjects (P<0.01). There was no difference in plasma leptin levels between nonobese diabetic subjects and controls. Obese diabetic subjects had significantly higher plasma leptin, TBARS, and insulin levels and significantly lower plasma zinc levels than nonobese diabetic subjects (for each comparison; P<0.01). The univariate and multivariate analyses demonstrated a significant positive correlation between leptin and body mass index (P<0.01) and insulin (P<0.01), and a significant negative correlation between leptin and zinc in obese subjects. Additionally, TBARS levels was positive correlated with insulin and negative correlated with zinc in obese diabetic subjects. We conclude that zinc may be a mediator of the effects of leptin, although the detailed mechanism is still unknown and requires further investigation. Free radical induced mechanism(s) may be involved in this process.

The altered hemorheologic parameters in thromboangiitis obliterans: a new insight.

The hemorheologic abnormalities were studied in patients suffering from thromboangiitis obliterans (TAO) and compared findings with those of patients suffering from critical limb ischemia because of peripheral atherosclerotic obstructive disease (PAOD) and the healthy subjects. This prospective study included 20 patients with TAO, 20 patients with PAOD, and 20 healthy subjects. Hematocrit, plasma fibrinogen, red blood cell rigidity, and blood and plasma viscosity were determined in all groups. Hematocrit values of the TAO group were significantly higher than those of the PAOD group and the control group (p < 0.001 and p < 0.001, respectively). BV at a shear rate of 6.00 sec(-1) was higher in TAO than the PAOD and control (p < 0.01 and p < 0.05, respectively). BV at a shear rate of 225 sec(-1) was higher in the TAO and PAOD than the control (p < 0.001 and p < 0.001, respectively) and also PAOD values were higher than the TAO (p < 0.05). RBC rigidity was higher in the TAO and PAOD than the control (p < 0.001 and p < 0.05, respectively) and these values were higher in PAOD than the TAO (p < 0.05). These results show the impairment in hemorheologic parameters in TAO may provide a new insight into the treatment of patients with TAO in the active period of the disease.

The effects of cholesterol levels on hemorheological parameters in diabetic patients.

Red blood cell (RBC) deformability is an important hemorheological parameter to determine the passage of RBC through narrow capillaries and the reduction of blood viscosity under high shear rates. Although it has been substantial evidence that diabetes mellitus (DM) and hypercholesterolemia increase the risk of coronary heart disease, the mechanism is unclear. In this study the relationship between hemorheological parameters and plasma cholesterol in type 2 diabetic patients (n=55, mean age 43.4+/-9.2 years) was examined. Type 2 diabetic patients were classified as normocholesterolemic (n=25; cholesterol < or = 200 mg/dl) and hypercholesteroloemic (n=30; cholesterol > 200 mg/dl) subgroups. Hypercholesterolemic type 2 diabetic patients had the highest blood and plasma viscosity and the lowest RBC deformability. The results were significantly different from normocholesterolemic type 2 diabetic patients (p<0.001). Our data suggest that elevated plasma cholesterol may impair RBC deformability and increase in blood and plasma viscosity by an additional effect to hyperglycemia in type 2 diabetic patients.

Comparison of plasma viscosity as a marker of endothelial dysfunction with nitric oxide and asymmetric dimethylarginine in subjects with dyslipidemia.

OBJECTIVE: In this study, we aimed to investigate the alterations in plasma viscosity and whether there was a relationship between plasma viscosity and endothelial dysfunction markers such as nitric oxide (NOx), asymmetric dimethylarginine (ADMA) and oxidized Low Density Lipoprotein (oxLDL) in dyslipidemic subjects. MATERIAL AND METHOD: 35 subjects with normolipidemia and 30 subjects with hyperlipidemia were involved in this study. Dyslipidemia was defined as total cholesterol levels ≥200 mg/dL and/or triglyceride level ≥150 mg/dL. Plasma total cholesterol, high density lipoprotein cholesterol, low density lipoprotein cholesterol, triglycerides, total protein, albumin levels and NOx were determined by enzymatic methods using commercial kits. Plasma ADMA concentrations and serum levels of total oxLDL were determined by ELISA. Plasma viscosity was measured by Harkness capillary viscometer. RESULT: Plasma viscosity, ADMA and oxLDL values were significantly higher in subjects with dyslipidemia than in subjects with normolipidemia. Plasma NOx concentration was decreased in dyslipidemic subjects compared to the normo-lipidemic subjects. We found that fibrinogen had no effect upon plasma viscosity in selected patients with dyslipidemia. CONCLUSION: Our results demonstrated that the rheological impairment of dyslipidemic patients was related with endothelial dysfunction and this was a possible cause of both micro and macrovascular complications. Therefore, as plasma viscosity is also a sensitive parameter, it can add useful information about the diagnosis and treatment of various disorders, and it should be utilized more frequently in clinical medicine.

The evaluation of plasma viscosity and endothelial dysfunction in smoking individuals.

BACKGROUND: Smoking is considered to be one the of risk factors effecting atherosclerosis which is associated the physical forces, biological and chemical stimuli occuring in vessel wall. The aim of this study is analysis of the biomechanical (plasma viscosity) and biochemical effect (nitric oxide, NOx; asymmetric dimethylarginine, ADMA) of smoking on endothelial function. METHODS: One hundred-twenty two individuals were divided into three groups according to their smoking status. Plasma viscosity was measured by Harkness Capillary Viscometer. Plasma NOx level was determined by enzymatic methods using commercial kits. ADMA concentration was determined by Elisa Plasma Assay and and physiologic spirometric and arterial gas parameters and pulmonary blood flow rate (PBFR) were measured. RESULTS: Viscosity variables of former smokers were significantly higher than those of non-smokers (p < 0.001). NOx levels were found to be statistically significantly higher when compared with current smokers and non-smokers (p < 0.001), and former smokers and non-smokers (p < 0.05). There was a higher fibrinogen levels in current smokers (p < 0.05) than smokers. CONCLUSIONS: Smoking increases the plasma viscosity that may lead endothelial damage. Plasma viscosity plays an important role as a biophysical mechanical marker on the behalf of hemodynamics. Biochemical markers, NOx and ADMA may show this damage, however, we observed that plasma viscosity can be consistent with biochemical markers. Thus, plasma viscosity may be useful for diagnosis, treatment and follow-up of the patients.

Blood viscosity as a forgotten factor and its effect on pulmonary flow.

BACKGROUND: The effect of smoking on blood viscosity is widely known. There are, however, few studies on the effect of blood viscosity on pulmonary circulation. METHODS: We aimed to observe the relationship between blood viscosity and pulmonary circulation among smokers and non-smokers. The study comprised 114 subjects in three groups: group 1, ex-smokers; group 2, smoked at least 10 packs/year and still smoking; group 3, never smoked. Blood viscosity (BV), pulmonary blood flow (PBF), and right ventricular systolic pressure (RVSP) were measured in all subjects. RESULTS: PBF was significantly lower in group 1 compared with group 3 (p < 0.05). BV in group 1 was significantly higher than group 3 (p < 0.05) while BV in group 2 was significantly higher than group 3 (p < 0.05). PBF in group 2 was significantly lower than group 3 (p = 0.01). CONCLUSIONS: We believe that BV is a significant and forgotten factor that plays an important role in pulmonary and cardiovascular diseases. BV may affect PF even during the course of smoking, and before the clinical onset of chronic obstructive pulmonary disease (COPD). Therefore, individuals at risk of pulmonary hypertension could be detected earlier with a simple blood test.

The effect of ß-glucan on trace element levels in intra-abdominal sepsis in rats.

Sepsis is associated with the development of progressive damage in multiple organ systems. The beneficial effect of glucans has been attributed to modulation of immune function and enhances defense against bacterial, viral, fungal, and parasitic infections. The aim of this study was to investigate the putative protective effect of ß-glucan on changes of trace element levels in various tissues after experimental sepsis in rats. Sepsis was induced by cecal ligation and perforation (CLP) in 28 male Wistar albino rats. To evaluate this, rats were divided into four groups as sham operated, ß-glucan treated sham operated, CLP, and ß-glucan treated CLP. Sixteen hours after operation, rats were decapitated and zinc (Zn) and copper (Cu) levels were determined in the liver, kidney, heart, diaphragm, and lung tissues. The results demonstrate that sepsis significantly decreased zinc and copper levels of all tissues. The decrease in tissue zinc and copper levels demonstrates the role of trace elements in sepsis-induced tissue damage. Our results indicated that ß-glucan administration did not return the zinc and copper levels to the control group level, and it seems likely that the given dose of ß-glucan was insufficient to prevent sepsis-induced organ injury.

Effects of hemorheological factors on the development of hypertension in diabetic children.

The aim of this study was to investigate the effects of hemorheological factors on the development of hypertension in diabetic children without retinopathy and persistent microalbuminuria. Arterial blood pressures were measured in 46 diabetic children and were compared with those of 29 healthy non-obese and 32 obese age- and sex-matched children. Higher systolic (SBP) (109.0 +/- 13.0 mmHg) and diastolic blood pressure (DBP) (74.3 +/- 9.5 mmHg) were obtained in diabetics (independent of age, sex, duration, and control degree of diabetes), when compared with non-obese children (SBP: 97.9 +/- 10.3 mmHg, DBP: 74.3 +/- 9.5 mmHg; p < 0.01, p < 0.05, respectively). No significant DBP and SBP difference was found between diabetics and obese children. When compared with non-obese children, blood viscosity, plasma viscosity, serum viscosity, serum albumin, and plasma fibrinogen values were found elevated in diabetics and were correlated with SBP and DBP. Serum haptoglobin levels and lipid profile were normal. The multivariate discriminant analysis demonstrated plasma viscosity and fibrinogen to be the most important variables related to the development of hypertension. The results of this study revealed that: (1) arterial blood pressures are high in diabetic patients independent of age, sex, duration of diabetes, control degree of diabetes, and lipid profiles; (2) arterial blood pressure levels in diabetic children are affected primarily from changes of plasma viscosity and fibrinogen; and (3) a common mechanism might play a role in the pathogenesis of hypertension in obese and diabetic children.