Polymorphonuclear leukocyte membrane fluidity and cytosolic Ca2+ content at baseline and after activation in essential hypertension

Cytosolic Ca2+Content and Membrane Fluidity of Platelets and Polymorphonuclear Leucocytes in Diabetes Mellitus

Considering the role played by platelets and leucocytes in diabetic disease and keeping in mind the strong correlation between functional and metabolic aspects that characterizes this clinical condition, we evaluated, in two groups of diabetics, respectively the platelet and polymorphonuclear (PMN) cytosolic Ca2+ content (employing the fluorescent probe Fura 2-AM) and membrane fluidity (using the fluorescent probe TMA-DPH and considering the fluorescence polarization degree, inversely related to the membrane fluidity). From the obtained results, it is evident that the platelet cytosolic Ca2+ content does not distinguish normals from diabetics of type 1 and 2; the platelet membrane fluidity …

Profilo emoreologico nell'infarto miocardico giovanile: nuove acquisizioni

Gelatinases and their tissue inhibitors in metabolic syndrome

Polymorphonuclear leukocyte integrins in chronic renal failure

Patients with chronic renal failure (CRF), in comparison with general population, show a higher cardiovascular mortality, not fully explained by the "traditional" risk factors. Among the new factors that have been hypothesized, leukocytes might play an important role. In a group of patients with mild CRF we determined, at baseline and after in vitro activation with 4-phorbol-12-myristate-13-acetate (PMA) and N-formyl-methionyl-leucyl-phenylalanine (fMLP), the polymorphonuclear leukocytes (PMN) beta2-integrin pattern (CD11a, CD11b, CD11c and CD18) by using indirect immunofluorescence with a flow cytometer. At baseline we observed an increase in the phenotypical expression of CD11b, CD11c and…

An assessment of the hemorheological profile in patients with subclinical carotid atherosclerosis divided in relation to the number of cardiovascular risk factors and different degrees of insulin resistance.

We present a cohort of 100 subjects [43 men and 57 women; median age 66.00(25)] who were tested using carotid ultrasound to identify subclinical carotid atherosclerosis (SCA). We have evaluated the behaviour of whole blood viscosity (WBV) at high (450 s–1) and low (0.51 s–1) shear rates, plasma viscosity (450–1), hematocrit and mean erythrocyte aggregation. When compared to normal control subjects, using the Mann-Whitney test, we observed in SCA patients a significant increase in WBV only. The results were substantial after having divided the SCA subjects according to the cardiovascular risk factors (CRFs) and the degree of insulin resistance; the research was performed using two surrogate …

Diabetes mellitus: evaluation of erythrocyte and polymorphonuclear leukocyte rheology

Aim: To explore red blood cells (RBC) and leukocyte rheology, that may be relevant in the pathophysiology of diabetes mellitus. Methods and Results: Significant alterations have been observed in RBC behaviour using several filtration techniques, but the exploration of RBC deformability by laser diffractometry did not show any abnormality. We have also employed microrheological methods based on fluorescence spectroscopy: membrane microviscosity was evaluated in ghosts, while in intact RBC we explored the membrane polarity gradient using fluorescent fatty acids, the phospholipid and protein lateral mobility using respectively pyrene and pyrene-3-maleimide. Alterations emerged only using the l…

Platelet membrane fluidity, platelet membrane lipid pattern and platelet cytosolic Ca2+ content in subjects with vascular atherosclerotic disease

In a group of subjects with vascular atherosclerotic disease (V AD) we examined the platelet membrane fluidity (obtained marking intact resting platelets with TMA-DPH), the platelet membrane cholesteroVphospholipid ratio (CIPL using column chromatography), the platelet membrane individual phospholipids (employing the thin layer chromatography) and the platelet cytosolic Ca2+ content (evaluated marking intact resting platelets with Fura 2-AM). From the obtained data, it is evident that platelet membrane fluidity differentiates normals from V AD subjects. Platelet membrane lipid pattern (CIPL and individual phospholipids) and cytosolic Ca2+ content do not discriminate normals from V AD subjec…

Nitric oxide metabolites, leukocyte activation markers and oxidative status in dialyzed subjects.

<i>Aims:</i> Our purpose was to evaluate, in a group of 42 end-stage renal disease patients who regularly undergo hemodialysis, some indexes of leukocyte activation, nitric oxide metabolites (NOx) and other parameters that reflect the oxidative stress before and after a standard hemodialysis session. <i>Methods:</i> Elastase and myeloperoxidase (MPO) were determined by means of ELISA. The NO production was evaluated by a micromethod which measures the concentration of NOx. The oxidation of polyunsaturated fatty acids was evaluated in plasma by detection of thiobarbituric acid-reactive substances (TBARS). Total antioxidant status (TAS) was obtained using spectrophotom…

Leucocyte Rheology at Baseline and after Activation in Post-Phlebitic Syndrome

Objective: To evaluate leucocyte rheology, expressed as leucocyte filtration, polymorphonuclear leucocyte (PMN) membrane fluidity and cytosolic Ca2+ concentration in subjects with post-phlebitic leg syndrome (PPS). Methods: In 22 subjects with PPS we determined leucocyte filtration [unfractionated, mononuclear (MN) and PMN cells], employing the St George Filtrometer, PMN membrane fluidity using the fluorescent probe 1-[4-(trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH) and PMN cytosolic Ca2+ concentration using the fluorescent probe Fura 2-AM. Subsequently we determined the same PMN parameters after in vitro activation with 4-phorbol 12-myristate 13-acetate (PMA) and N-formyl-met…

Vascular atherosclerotic disease: Evaluation of white blood cell rheology and metabolism after acute intravenous administration of defibrotide

Defibrotide is a single-strand polydeoxyribonucleotide, obtained by controlled depolymerization of DNA extracted from mammalian organs. It stimulates vascular prostacyclin production and secretion (1-3) and enhances fibrinolysis (1, 4-6). Several clinical trials have evidentiated that defibrotide is effective in the prophylaxis of deep vein thrombosis and in the treatment of peripheral obliterative arterial disease (for a review see ref. 7). There are several reports concerning, in vitro and in vivo, the dose-dependent activity of defibrotide on the polymorphonuclear leucocytes (PMN). From these results, the molecule influences the PMN adhesivity (8, 9) and especially PMN activation, with s…

Membrane fluidity, membrane lipid pattern, and cytosolic Ca2+ content in platelets from a group of type II diabetic patients with macrovascular complications

OBJECTIVE To evaluate platelet membrane fluidity and some platelet metabolic parameters in type II diabetic patients with macrovascular complications. RESEARCH DESIGN AND METHODS In a group of 21 type II diabetic patients with macrovascular complications, we evaluated platelet membrane fluidity [marking intact resting platelets with the fluorescent probe 1,4-(trimethylamino)-phenyl-4-phenylhexatriene (TMA-DPH)], platelet membrane lipid pattern (cholesterol :phospholipid [C:PL] ratio and individual phospholipids), and platelet cytosolic Ca2+ content (marking intact resting platelets with the fluorescent probe Fura 2AM). RESULTS Platelet membrane fluidity is decreased in type II diabetic pat…

Oxidative status in metabolic syndrome

Protein carbonyl groups in trained subjects.

The purpose of this research was to evaluate the plasma protein carbonyl groups (PC) in 81 trained subjects (TS) who practiced regular, non professional physical activity. They were divided into three groups according to the type of sport they practiced (endurance, mixed or power). On fasting venous blood we examined the PC groups employing an enzyme-linked immunosorbent assay (ELISA) kit, in which 2,4-dinitrophenyl-hydrazine reacts with the PC forming a stable hydrazone product. In the whole group of TS a significant decrease in PC was present, in comparison with sedentary controls (SC). Dividing TS into groups, we observed a decreased PC concentration in those practicing endurance and mix…

Lipid peroxidation and protein oxidation are related to the severity of OSAS

OBJECTIVE: Obstructive sleep apnea syndrome (OSAS) is associated with elevated cardiovascular morbidity and mortality. Considering that oxidative stress is involved in endothelial dysfunction and atherosclerosis development, our aim was to examine lipid peroxidation and protein oxidation, two parameters of oxidative status, in a group of subjects with OSAS. PATIENTS AND METHODS: We consecutively enrolled 48 patients (36 men and 12 women; mean age 49.7±14.6 yrs) with OSAS, subsequently subdivided according to the apnea/hy-popnea index (AHI) value in two subgroups: Low (L= 21 subjects with AHI30). We examined lipid peroxidation, expressed as TBARS, and protein oxidation, measured as carbonyl …

Polymorphonuclear leukocyte membrane fluidity and cytosolic Ca2+ concentration in diabetes mellitus.

We evaluated polymorphonuclear membrane (PMN) fluidity in 32 subjects with type 1 diabetes mellitus, 38 subjects with type 2 diabetes mellitus and 38 normal control subjects, by marking intact and unstimulated PMN cells with the fluorescent probe 1-[4-(trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH). We also evaluated PMN cytosolic Ca2+ content by marking intact and unstimulated PMN cells with the fluorescent probe Fura 2-AM. PMN membrane fluidity differentiated normal subjects from type 1 and 2 diabetic subjects. The PMN cytosolic Ca2+ concentration did not discriminate type 1 and 2 diabetic subjects from normal control subjects. No statistical correlation was found between PMN …

Acute Ischemic Stroke

Background and Purpose —Several reports have considered the role of systemic leukocytes in acute ischemic stroke (AIS). Initially, greater attention was focused on the leukocyte count and subsequently on their adhesiveness, aggregation, rheology, and activation. The aim of this study was the evaluation of certain polymorphonuclear leukocyte (PMN) parameters, reflecting their rheology and activation, in subjects with AIS. Methods —In a group of 19 subjects with AIS and in a control group of 18 subjects with asymptomatic vascular atherosclerotic disease, we evaluated the PMN membrane fluidity and cytosolic Ca 2+ concentration at baseline and after in vitro chemotactic activation with 4-phorb…

Relationship between heart rate variability and repeated sprint exercise in young soccer players

Aim: The sport of soccer is characterized by neuromuscular performance resulting in short bursts interspersed by brief recovery periods over an extended period of time. Several studies suggest that the cardiac autonomic nervous system (ANS) assessed by heart rate variability (HRV) may provide useful information regarding the functional adaptations to training stimulus. Although HRV has often been associated with aerobic power and cardiovascular fitness, the influence of sympathovagal balance on neuromuscular performance in young athletes is not well assessed. The aim of this study was to assess the influence of the autonomic system on the performance of repeated sprints, involving predomina…

Hemorheologic profile of hyperlipidemic patients treated with gemfibrozil

Abstract We measured the plasma lipid levels and the macrorheologic (whole-blood, plasma, and serum viscosity levels; fibrinogen; hematocrit; mean erythrocyte aggregation; and whole-blood filterability) and microrheologic (erythrocyte membrane fluidity and red cell membrane protein lateral mobility) determinants in a group of 19 hyperlipidemic patients (Fredrickson's classification phenotype IIa, 12 patients; phenotype IV, 4; phenotype IIb, 3) at baseline and after 45 and 90 days of treatment with gemfibrozil (900 mg orally once a day). At baseline we noted statistically significant increases in plasma viscosity level, fibrinogen, and mean erythrocyte aggregation, as well as a statistically…

Red Cell Filterability and Erythrocyte Membrane Microviscosity during Ticlopidine Treatment

The effects of 10 days' treatment with 500 mg/day ticlopidine on the filterability of red blood cells in suspension and on the microviscosity of the red blood cell membrane was evaluated in 11 patients with vascular atherosclerosis. The results show ticlopidine to be effective in influencing the rheological measures of red cell filterability and membrane microviscosity: filterability was increased and microviscosity was decreased. Ticlopidine also decreased the extent of fluorescence polarization. These results are discussed regarding possible mechanisms of action of ticlopidine.

Erythrocyte deformability, plasma lipid peroxidation and plasma protein oxidation in a group of OSAS subjects

Considering that obstructive sleep apnea syndrome (OSAS) is usually associated with endothelial dysfunction, atherosclerosis and cardiovascular disorders, our aim was to examine the erythrocyte deformability and the oxidative status in a group of OSAS subjects. We consecutively enrolled 48 subjects with OSAS defined after a 1-night cardiorespiratory sleep study, subsequently subdivided according to the apnea/hypopnea index (AHI) value in two subgroups: Low (L = 21 subjects with AHI30) and High (H = 27 subjects with AHI30). We evaluated the erythrocyte deformability, expressed as elongation index (EI) and the parameters of the oxidative status, such as lipid peroxidation (expressed as thioba…

Polymorphonuclear leukocyte cytosolic Ca2+ content in non-dialyzed subjects with chronic renal failure

Rheological and metabolic leucocyte determinants in diabetes mellitus

In diabetics of type I and 2 we examined, in resting white blood cells (WBC), the filtration parameters (Initial Relative Flow Rate - lRFR, Clogging Rate - CR) employing the St. George Filtrometer, the polymorphonuclear cells (PMN) membrane fluidity, the PMN cytosolic Ca2+ content and the PMN membrane cholesterol/phospholipid ratio (C/PL). From the obtained data, it is evident that, while the lRFR of unfractionated WBC distinguishes normals from diabetics of type 1 and 2, the fIltration parameters of the PMN and mononuclear cells (MN) do not show any significant difference. PMN membrane fluidity, PMN cytosolic Ca2+ content and PMN C/PL do not discriminate normals from diabetics of type 1 an…

Short-Term Prognosis of Juvenile Myocardial Infarction

In our early research regarding the hemorheological pattern in patients with acute myocardial infarction (AMI) with a mean age of 61.45 + 10.99 years, we showed that the major hemorheological parameters were almost normalized 2 weeks after the acute event. In the last decade, we focused on hemorheological parameters in juvenile myocardial infarction (JMI), defined as AMI in patients aged 45 years, in the ‘‘Sicilian study on juvenile myocardial infarction’’. Juvenile myocardial infarction is responsible for 2% to 10% of all cases with AMI in different surveys. Juvenile myocardial infarction presents a typical pattern of risk factors and shows clinical, angiographic, and prognostic characteri…

Interrelazioni tra disfunzione endoteliale e stress ossidativo; valutazione della perossidazione lipidica in particolari quadri clinici

Il termine "disfunzione endoteliale" identifica una condizione patologica caratterizzata da ridotta vasodilatazione e da attività procoagulante e pro-infiammatoria, correlata ad una ridotta biodisponibilità di ossido nitrico (NO), ad un eccesso ossidativo e ad un'aumentata azione dell'endotelian 1 (ET-1).

Red Cell Membrane Protein Lateral Mobility in Diabetes Mellitus

In a group of 24 diabetics subdivided for type, we evaluated the red cell membrane protein lateral mobility marking intact red cells with pyrene-3-maleimide (3-PM) and calculating the dimer to monomer fluorescence intensity ratio (Iex/Im). The same fluorescent parameter was determined in a group of 13 normal controls. From the obtained data, it is evident that the red cell membrane protein lateral mobility clearly discriminates normals from diabetics of type 1 and 2. In normals and in diabetics of type 1 and 2 no relationship is present between this fluorescent determinant and the glycometabolic parameters (FBGL and HbA1c); considering all the diabetics, a negative relationship is evident b…

Polymorphonuclear leukocyte membrane fluidity before and after activation in subjects with insulin resistance.

The aim of this research was the evaluation of polymorphonuclear leukocyte (PMN) membrane fluidity in subjects with insulin resistance. Insulin sensitivity, in fact, may be influenced by plasma membrane fluidity. We enrolled 19 subjects with insulin resistance previously demonstrated during an euglycemic hyperinsulinemic clamp. PMN membrane fluidity was studied by labeling intact cells with the fluorescent probe 1-[4-(trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene and calculating the fluorescence polarization degree. The measurement was made before and after incubation of PMNs with two activating agents: 4-phorbol 12-myristate 13-acetate (PMA) and N-formyl-methionyl-leucyl-phenylalanine (…

Role of genetic polymorphisms in myocardial infarction at young age

Acute myocardial infarction (AMI) in young adult presents a typical pattern of risk factors, clinical, angiographic and prognostic characteristics. In the last years we demonstrated that hemorheological profile is altered in these patients in a persistent way and independently of the number of risk factors and of the extent of coronary lesions. Thus, the hyperviscosity syndrome following AMI could be considered an intrinsic characteristic of these patients. Consequently it is possible to hypothesise the presence of a genetic background at the origin of this predisposition. If this background is able to influence the risk of ischemic heart disease, this should be particularly evident in youn…

Oxidative status in nondiabetic middle-aged subjects with metabolic syndrome: preliminary data.

Diabetes mellitus: oxidative stress and wine.

This review focuses on the link between diabetes mellitus and oxidative stress and, in particular, on the role that moderate wine consumption may play in preventing diabetic complications and the onset of diabetes. With this aim, a search of PubMed was carried out for literature published up to March 2003. In diabetes mellitus, oxidative stress results both from exposure to hyperglycaemia through glycoxidation and sorbitol system activation, and from functional limitation of the hexose monophosphate shunt, leading to a decrease in glutathione synthesis. Oxidative stress alters the plasma lipoprotein profile (particularly low-density lipoproteins), the coagulative parameters (with an increas…

Platelet membrane fluidity and platelet membrane lipid pattern in several clinical conditions

Polymorphonuclear leucocyte rheology and cytosolic Ca2+ content after activation in chronic renal failure

SUMMARY: We evaluated polymorphonuclear leucocyte (PMN) flow properties in patients with clinically stable chronic renal failure (CRF) and in control subjects at baseline and after activation with 4-phorbol 12-myristate 13-acetate (PMA) and N-formyl-methionyl-leucyl-phenylalanine (fMLP). Initial relative flow rate (IRFR) and clogging particles (CPs) were obtained using the St. George's Filtrometer, and PMN membrane fluidity was assessed by marking PMNs with 1-(4-(trimethylamino)phenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH). PMN cytosolic Ca2+ concentration was determined by marking PMNs with Fura 2-AM. At baseline, CRF patients showed a significant increase only in PMN cytosolic Ca2+ content.…

Effect of nimodipine on rheologic parameters in patients with chronic cerebrovascular disease

Abstract Sixteen patients with chronic cerebrovascular disease were treated with a monotherapeutic regimen of nimodipine 30 mg orally three times a day. At baseline and after 45 and 90 days of therapy, the patients' whole blood filterability, erythrocyte membrane fluidity, red blood cell membrane protein lateral mobility, and red blood cell Ca 2+ content (total and cytosolic) were evaluated. After 45 days, an increase in whole blood filterability and a decrease in the cytosolic red cell Ca 2+ content was seen. At the end of treatment, whole blood filterability, erythrocyte membrane fluidity, and red cell membrane protein lateral mobility had increased, compared with baseline values, while t…

Diabetes mellitus: mean erythrocyte aggregation, glycometabolic pattern, red cell Ca2+ content, and erythrocyte membrane dynamic properties.

Nitric oxide metabolites and oxidative stress in mild essential hypertension

Many papers have showed non univocal data about oxidative stress status and nitric oxide metabolites in essential hypertension. Considering this preamble we examined the total antioxidant status (TAS), the lipid peroxidation (LP), expressed as thiobarbituric acid-reactive substances (TBARS), the stable end products of nitric oxide (NOx) and LP/NOx ratio in 25 subjects with untreated mild essential hypertension. The obtained data show a significant increase in TBARS (p < 0.001) and NOx (p < 0.001) in hypertensives and no variation in TAS and in TBARS/NOx ratio. None of these parameters was statistically related to the metabolic parameters or to the blood pressure values. The high level of li…

Red Cell Membrane Protein Lateral Mobility in Vascular Atherosclerotic Disease: Preliminary Report

Beta-Thromboglobulin and Platelet Factor 4 in Juvenile Myocardial Infarction

Leukocyte activation markers and oxidative status in chronic kidney disease

Behaviour of the neutrophil to lymphocyte ratio in young subjects with acute myocardial infarction

In the last years the neutrophil to lymphocyte ratio (NLR) has been examined in cardiovascular disorders and in particular in coronary artery disease and acute myocardial infarction (AMI). Now we examined this parameter in subjects with juvenile myocardial infarction at the initial stage and after 3 and 12 months. We enrolled 123 young subjects (112 men and 11 women, mean age 39.4 ± 5.8 yrs) with AMI. The time interval between the AMI onset and the investigation was 13 ± 7 days. The mean value of NLR observed in young AMI subjects was significantly increased compared to normal controls (N = 1.817 ± 0.711; young AMI subjects = 2.376 ± 0.873, p <  0.0001). NLR does not discriminate STEMI (2.4…

Polymorphonuclear leukocyte integrin pattern, at baseline and after activation, in type 2 diabetic subjects with macrovascular complications.

In vascular atherosclerotic disease and in diabetes mellitus few studies have evaluated the polymorphonuclear leukocyte (PMN) adhesion molecule pattern. In this study we examined the PMN integrin expression at baseline and after activation in controls and type 2 diabetic subjects with macrovascular complications (MVC). We enrolled 21 subjects with type 2 diabetes mellitus and macrovascular complications, localized in peripheral, coronary and cerebral sites. The patients had peripheral occlusive arterial disease, chronic cerebrovascular disease or coronary heart disease. We evaluated the expression of some PMN integrins (CD11a, CD11b, CD11c, CD18), using flow cytofluorimetry, at baseline and…

Chronic renal failure: leukocyte rheology and polymorphonuclear cytosolic Ca2+ concentration.

In a group of subjects with chronic renal failure (CRF), we evaluated the leukocyte rheological activity, expressed as the filtration rate, the polymorphonuclear (PMN) membrane fluidity and the PMN cytosolic Ca2+ content. Using the St. George's Filtrometer, we determined the filtration rate of unfractionated, mononuclear and PMN leukocytes. Using the fluorescent probe 1.4-(trimethylamino)-phenyl-4-phenyl-hexatriene (TMA-DPH), we examined the PMN membrane fluidity and, using the Fura 2-AM, the PMN cytosolic Ca2+ content. From the results obtained, it is evident that only the initial relative flow rate of unfractionated leukocytes was significantly reduced in subjects with CRF, while the filt…

Protein carbonyl groups in trained subjects before and after a cardiopulmonary test

Physical exercise influences the body's oxidative status. The modifications can involve lipids, proteins and nucleic acids, and different effects seem to be induced by regular and acute exercise respectively. We examined protein oxidation, expressed as concentration of protein carbonyl groups (PC), in trained subjects before (time 0), 10 min (time 1) and 24 hours (time 2) after a cardiopulmonary test performed on a cycloergometer. We enrolled 38 trained subjects (26 men and 12 women), subdivided in two groups (A1 and B1) of 19 subjects each, according to the median value of VO2max, and in two groups (A2 and B2) of 19 subjects each, according to the median value of PC at baseline. PC concent…

Fluidity and cytosolic Ca2+ concentration of circulating polymorphonuclear leukocytes at baseline in some chronic and acute clinical conditions: review of our survey

Abstract. Objective: In this mini-review we describe the behavior of polymorphonuclear leukocyte (PMN) membrane fluidity and of PMN cytosolic Ca2+ concentration in some chronic and acute clinical conditions. Methods: PMN membrane fluidity was evaluated employing the fluorescent probe Fura-2AM, and PMN cytosolic Ca2+ concentration was evaluated using the fluorescent probe TMA-DPH. Results: From the determination of these two parameters investigated on resting PMNs, an almost constant increase in PMN cytosolic Ca2+ concentration in chronic clinical conditions, such as vascular atherosclerotic disease with and without diabetes mellitus, essential hypertension, chronic kidney disease, and diabe…

Platelet activation markers in long-term follow-up of young subjects with acute myocardial infarction

Methylenetetrahydrofolate reductase mutation in subjects with abdominal aortic aneurysm subdivided for age.

Myocardial infarction in Young adults: risk factors, clinical characteristics and prognosis according to our experience

Objectives. Myocardial infarction is a relatively unusual phenomenon in young subjects. The aim if this work is to characterize the risk profile and factors influencing outcomes of these patients since it makes possible to manage prevention interventions. Patients and Methods. We examined cardiovascular risk factors, clinical presentation, angiographic picture and outcome of a group of young patients hospitalized for a myocardial infarction. We enrolled 121 young patients consecutively admitted to our hospital for a myocardial infarction and examined them not only at the initial stage, but also after 3 months and one year; finally a long-term telephonic follow up was performed, when possibl…

Hemorheological parameters in Monoclonal Gammopathy of Undetermined Significance (MGUS).

There is scarcity of information about the hemorheological pattern in subjects with Monoclonal Gammopathy of Undetermined Significance (MGUS). This preliminary research is focused on the behaviour of whole-blood and plasma viscosity, haematocrit and erythrocyte deformability in the above clinical condition. We enrolled 21 MGUS subjects (10 women and 11 men; mean age 66.4 ± 11.6 years). In fasting venous blood we examined whole-blood and plasma viscosity at high and low shear rates, haematocrit, the ratios between whole-blood viscosity (at high and low shear rate) and haematocrit × 100, the ratio between plasma viscosity at low and high shear rate, and the erythrocyte deformability expressed…

Polymorphonuclear leukocyte integrins in deep venous thrombosis

The polymorphonuclear leukocytes (PMN) have a role in the pathophysiology of deep venous thrombosis (DVT). We examined the phenotypical expression of PMN beta2-integrins (CD l l a, CDl l b, CD 11c) in a group of 19 subjects with leg DVT. PMN cells were incubated with fluorescent monoclonal antibodies against CD11a, CD11b, CD11c, and the evaluation was made by flow cytofluorimetry. The same integrins were determined after in vitro activation with 4-phorbol 12-myristate 13-acetate (PMA) and N-formylmethionyl-leucyl-phenylalanine (fMLP). In DVT subjects, at baseline, the phenotypical expression of CD11b was decreased and that of CD11c increased when compared with normal controls. In normal su…

Deep venous thrombosis: Behaviour of the polymorphonuclear leukocyte integrin pattern at baseline and after in vitro activation

In a group of 18 subjects with acute deep venous thrombosis (DVT), evidenced by clinical examination and echo-color-Doppler, we examined the phenotypical expression of the polymorphonuclear leukocyte (PMN) beta2-integrins (CD11a, CD11b, CD11c, CD18), obtained by using a flow cytofluorimeter. The evaluation was performed before and after in vitro activation (prolonged for 5 and 15 minutes) with 4-phorbol 12-myristate 13-acetate (PMA) and N-formyl-methionyl-leucyl-phenylalanine (fMLP). In DVT subjects, at baseline, the phenotypical expression of CD11b was decreased and that of CD11c was increased when compared with normal controls; no difference was found in CD11a and CD18 expression. In norm…

Polymorphonuclear leukocyte membrane fluidity, at baseline and after in vitro activation, in obesity with or without diabetes mellitus

We studied a group of 28 obese subjects (mean age 38.2+/-13.5 years, body mass index 35.0+/-5.6 kg/m2) with insulin resistance demonstrated employing an euglycemic hyperinsulinemic clamp, subdivided into a subgroup with normal glucose tolerance (NGT) and a subgroup with type 2 diabetes mellitus (DM). We examined the polymorphonuclear leukocyte (PMN) membrane fluidity at baseline and after activation with 4-phorbol 12-myristate 13-acetate (PMA) or N-formyl-methionyl-leucylphenylalanine (fMLP). At baseline, PMN membrane fluidity was significantly decreased in both subgroups compared to normals. In obese subjects with NGT no correlation was found between this PMN determinant and the parameters…

Granulocyte cytosolic calcium in type 2 diabetes

Plasma markers of platelet and polymorphonuclear leukocyte activation in young adults with acute myocardial infarction

Our goal was to evaluate some plasma markers of platelet and polymorphonuclear leukocyte (PMN) activation in a group of young adults with acute myocardial infarction (AMI) at the initial stage and after three months. We enrolled 49 AMI subjects aged<45 years and examined plasmatic levels of platelet factor 4 (PF4), β-thromboglobulin (β-TG), elastase and myeloperoxidase (MPO) using ELISA methods. PF4 and β-TG were increased, compared to control subjects, both at the initial stage and after 3 months. In control subjects and in AMI patients, at both times of observation, there was a significant and positive correlation between the two platelet parameters, while no correlation was present betwe…

Haematocrit in metabolic syndrome

Erythrocyte deformability and hemorheological profile in multiple myeloma

The hemorheological profile in multiple myeloma (MM) has been extensively studied. Our investigation regarded the behavior of whole-blood viscosity, plasma viscosity and erythrocyte deformability in MM. We enrolled 24 MM patients; 13 of them had been recently diagnosed and were at the initial stage of therapy, 6 were on consolidation/conservation therapy and 5 had achieved a complete remission. On fasting venous blood we evaluated whole-blood and plasma viscosity at high and low shear rates, haematocrit, the ratios between whole-blood viscosity (at high and low shear rate) and haematocrit×100, the ratio between plasma viscosity at low and high shear rate and the erythrocyte deformability ex…

Polymorphonuclear leukocyte
integrin pattern in acute
ischemic stroke

Haemorheological profile in congenital afibrinogenemia and in congenital dysfibrinogenemia: A clinical case report

Although the inherited quantitative and qualitative disorders of fibrinogen are rare, in the course of time patients may develop complications including episodes of arterial and venous thrombosis. It can be useful to complete the laboratory assessment of these clinical conditions with the evaluation of the haemorheological profile. The data obtained from this study showed that congenital afibrinogenemia was characterized by a primary plasma hypoviscosity, whereas congenital dysfibrinogenemia by a primary plasma hyperviscosity. Both these haemorheological alterations may concur, with different mechanisms, to the pathogenesis of thrombotic vascular complications.

Protein oxidation in mild essential hypertension

An intensified oxidative stress has been associated with aging and many disease, including essential hypertension (EH) [6, 11, 16]. In EH an increased level of reactive oxygen and nitrogen species (RONS) may impair the bioavailability of nitric oxide (NO), by inducing its accelerated transformation in peroxynitrite [4]. Beside this, many other pathophysiological processes in EH may be influenced by RONS, considering their harmful effects on the structural and functional properties of lipids, proteins and nucleic acids. Some points need to be further investigated, such as the relationship between oxidative stress and the degree of hypertension, and identification of the most useful marker of…

Impairment of plasma nitric oxide availability in senescent healthy individuals: Apparent involvement of extracellular superoxide dismutase activity

To verify the potential involvement of the age-dependent modifications of EC-SOD activity in the impairment of plasma NO availability with advancing age, 40 healthy men divided into 4 age groups for the purpose of comparison (young: 27.4 +/- 1.5 years; middle: 50.8 +/- 2.2, years; old: 70.0 +/- 1.8 years; very old: 86.1 +/- 1.1 years) were enrolled in this study. Plasma samples were used for measurements of the stable end-product nitrite/nitrate (NOx), as an expression of NO availability, EC-SOD activity, thiobarbituric acid reactive substances (TBARS) as a marker of lipid peroxidation, low density lipoprotein (LDL) copper-mediated oxidation in vitro and total antioxidant capacity (TEAC). O…

Behaviour of the plasma concentration of gelatinases and their tissue inhibitors in subjects with venous leg ulcers.

Venous leg ulcers are common in subjects with chronic venous insufficiency. The increased intraluminal pressure causes alteration of the skin microcirculation, leukocyte activation and release of proteolytic enzymes leading to ulceration. An impaired expression and activity of matrix metalloproteases (MMPs) and their tissue inhibitors (TIMPs) might influence extracellular matrix degradation and deposition in chronic venous ulcers with the failure of the healing process. Our aim was to evaluate plasma concentration of gelatinases (MMP-2 and MMP-9) and their inhibitors (TIMP-1 and TIMP-2) in subjects with venous leg ulcers before and after the compression therapy. We enrolled 36 subjects (12 …

Study of the Correlations among Some Parameters of the Oxidative Status, Gelatinases, and Their Inhibitors in a Group of Subjects with Metabolic Syndrome

Our aim was to examine some parameters of oxidative status, gelatinases, and their inhibitors and to evaluate their interrelationships in subjects with metabolic syndrome (MS). We enrolled 65 MS subjects, subdivided according to the presence or not of diabetes mellitus. We examined lipid peroxidation (expressed as thiobarbituric acid reacting substances, TBARS), protein oxidation (expressed as carbonyl groups), nitric oxide metabolites (NOx), total antioxidant status (TAS), MMP-2, MMP-9, TIMP-1, and TIMP-2. We found that MS subjects, diabetics and nondiabetics, showed an increase in TBARS, PC, and NOx. A significant decrease in TAS was observed only in nondiabetic MS subjects in comparison …

Vascular atherosclerotic disease: Behaviour of the polymorphonuclear leukocyte (PMN) filtration parameters, PMN membrane fluidity and PMN cytosolic Ca2+ content after chemotactic activation

Chronic renal failure: oxidative stress, endothelial dysfunction and wine.

Atherosclerosis development is accelerated in chronic renal failure (CRF) and is the major cause of death in this clinical condition. An increased oxidative stress and an endothelial dysfunction, with their complex interrelationships, are relevant aspects of atherogenesis in CRF patients and might be targets for treatment. Many studies have underlined the cardiovascular protection provided by a moderate wine consumption. This beneficial effect is due to both alcohol and nonalcoholic components of wine including several phenolic molecules such as quercetin and resveratrol. Wine polyphenols have antioxidant properties and favorably influence endothelial function, in particular by stimulating …

Type II diabetics with macrovascular complications: polymorphonuclear leukocyte (PMN) filtration, PMN membrane fluidity and cytosolic Ca2+ content after activation.

We evaluated polymorphonuclear (PMN) filtration parameters, membrane fluidity and cytosolic Ca2+ content in 21 normal subjects and in 18 type II diabetics with macrovascular complications (MVC). Evaluations were carried out at baseline and after in vitro activation prolonged for 5 and 15 min. PMA (4-phorbol 12-myristate 13-acetate) and fMLP (N-formyl-methionyl-leucyl-phenylalanine) were used as stimulating agents. TMA-DPH (1-[4-(trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene) was used as fluorescent probe for the membrane fluidity tests and Fura 2-AM for the cytosolic Ca2+ content. A significant variation was evident in PMN filtration parameters at 5 and 15 min. No variation was present i…

Techniques to evaluate erythrocyte deformability in diabetes mellitus

Using several rheological techniques, we examined the erythrocyte deformability in different groups of diabetic subjects. The macrorheological techniques used for this evaluation were respectively whole-blood filtration, filtration of erythrocyte suspensions, polyviscosimetry and diffractometry. Whole-blood filterability, at a negative pressure of 20 cm water, was decreased in type 2 diabetics; no difference was evident at a negative pressure of 10 cm water. The filtration of erythrocyte suspensions at low haematocrit (5%) did not show differences between normal and diabetic subjects. The polyviscosimetry, which explores the filterability of erythrocyte suspensions at high haematocrit (80%)…

Pathophysiological and clinical aspects of malnutrition in chronic renal failure

AbstractKidney diseases are the ninth leading cause of death in the USA. In these patients cardiovascular mortality is greater than in the general population. This observation, not completely explained by the so-called ‘traditional’ cardiovascular risk factors, lead the authors to postulate other ‘emerging’ ones found in chronic renal failure patients. Among these new findings, nutritional status, considered as the balance existing between nutrient requirements and intakes, plays an important role for the development of cardiovascular diseases. In fact several nutritional parameters are widely known as pathophysiological determinants of cardiovascular disturbances, which are based on accele…

Leukocyte subtypes, gelatinases, and their tissue inhibitors in a group of subjects with asymptomatic carotid atherosclerosis

In a cohort of subjects with asymptomatic carotid atherosclerosis (ACA), we have evaluated the neutrophil and lymphocyte count and their ratio (NLR), the gelatinases (MMP-2 and MMP-9) and their tissue inhibitors (TIMP-1 and TIMP-2). At baseline, no difference was observed between ACA subjects and subject control group regarding neutrophil and lymphocyte count while was evident in ACA subjects a significant increase in MMP-2, MMP-9 and TIMP-2 associated to a significant decrease in TIMP-1. Dividing the ACA according to the number of cardiovascular risk factors (CRFs) we have observed an increase in lymphocyte count in the subgroup with 3–5 CRFs. Evaluating the leukocyte subtypes according to…

Pallidotomy improves quality of life in selected parkinsonian patients: an Italian report.

Pallidotomy has recently been reconfirmed as effective for otherwise intractable symptoms of Parkinson's disease. Nonetheless almost every aspect of its performance requires choices which are not fully established and may vary between centers. These include: 1) patient selection; 2) choice of imaging modality, 3) choice of anatomic landmarks for targeting the lesion, 4) choice of method for physiologic confirmation of location, 5) choice of lesion size and shape. We present two cases of pallidotomy procedures in Parkinsonian patients that in our knowledge are the first reported in Italy. Our experience and a careful review of the literature led to the following choices: 1) selection of Park…

Nitric oxide metabolites (nitrite and nitrate) in several clinical condition.

We determined the concentration of nitric oxide metabolites (NO2-+NO3-), expressed as NOx, in several clinical conditions. Regarding this, we have examined 25 subjects with arterial hypertension, 41 subjects with chronic kidney disease in conservative treatment, 106 subjects with metabolic syndrome subdivided according to the presence (n = 43) or not (n = 63) of diabetes mellitus, 48 subjects with obstructive sleep apnea syndrome (OSAS), 14 women with systemic sclerosis complicated with Raynaud's phenomenon, 42 dialyzed subjects and 105 young subjects with acute myocardial infarction (AMI). In subjects with arterial hypertension, chronic kidney disease, metabolic syndrome, systemic sclerosi…