Baldassare Canino

Immune-inflammatory markers and arterial stiffness indexes in subjects with acute ischemic stroke with and without metabolic syndrome

ObjectiveThe aim of our study was to evaluate the associations between arterial stiffness indexes and immune-inflammatory markers in subjects with acute ischemic stroke with and without metabolic syndrome.Materials/MethodsWe enrolled 130 patients with acute ischemic stroke and metabolic syndrome, 127 patients with acute ischemic stroke without metabolic syndrome and 120 control subjects without acute stroke. Applanation tonometry was used to record the augmentation index (Aix) and pulse wave velocity (PWV). We also evaluated plasma levels of C-reactive protein (CRP), Interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α), Interleukin-6 (IL-6) and Interleukin-10 (IL-10), E-selectin, P-selectin, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), von Willebrand Factor (vWF) plasma levels, tissue plasminogen activator (TPA) and plasminogen activator inhibitor-1 (PAI-1).ResultsIn subjects with acute ischemic stroke and metabolic syndrome we observed higher median plasma values of immuno-inflammatory markers. In acute ischemic stroke patients and metabolic syndrome in relation of each TOAST subtype we observed a more significant positive correlation between PWV and immuno-inflammatory markers.ConclusionsStroke subjects with acute ischemic stroke and metabolic syndrome showed a higher degree of immuno-inflammatory and arterial stiffness indexes possibly due to metabolic background of these types of patients that trigger a more intense immune-inflammatory activation irrespective of stroke subtype, whereas being related to stroke subtype in subjects without metabolic syndrome.

Granulocyte integrins before and after activation in acute ischaemic stroke.

We examined in 19 subjects with acute ischaemic stroke (AIS) the PMN integrin pattern (CD11a, CD11b, CD11c, CD18), using indirect immunofluorescence and adopting a flow cytometer, at baseline and during activation, prolonged for 5 and 15 min, with 4-phorbol 12-myristate 13-acetate (PMA). At baseline, an increase in the expression of CD11c and CD18 and a decrease in the CD11b were evident in AIS subjects compared to normals. After activation, we found in normals a constant and significant increase of all PMN adhesive molecules, while in AIS subjects, we found an increase in CD11b and CD18, a decrease in CD11a and no variation in CD11c. While the basal upregulation of CD11c and CD18 may depend on the PMN spontaneous activation or on the increase of cytokines, the decrease of CD11b may be due to its self-consumption. After activation, the decrease in CD11a noted in AIS may be related to its cleavage or to an altered integrin phosphorylation/dephosphorylation balance.

Lipid peroxidation and total antioxidant status in unprofessional athletes before and after a cardiopulmonary test.

We examined lipid peroxidation, expressed as thiobarbituric acid-reactive substances (TBARS) and total antioxidant status (TAS) before and after a cardiopulmonary test, in 20 sedentary controls and in 62 unprofessional male athletes subdivided into 3 subgroups. The first included subjects who practised endurance sports (14 cyclists and 9 endurance swimmers), the second subjects who practised mixed sports (6 basket players, 6 judoists, 8 water polo players) and the third group subjects who practised power sports (3 sprint runners, 4 weightlifters, 12 sprint swimmers). In the whole group of athletes an increase in TBARS and a decrease in TAS were present at baseline. Subdividing the whole group into three subgroups we observed an increase in TBARS in all and a decrease in TAS only in the endurance and mixed athletes. After the test, TBARS showed a more significant increase in controls compared to the whole group and each subgroup of athletes, while TAS increased only in the whole group and in those who practised mixed sports. In conclusion, at baseline in athletes the oxidative status shows a different behaviour compared to controls, while after the test the antioxidant protection is more marked and it may be related to an increase of antioxidant systems.

Polymorphonuclear Integrins, Membrane Fluidity, and Cytosolic Ca2+ Content After Activation in Essential Hypertension

The purpose of this research was to obtain further information about the role of polymorphonuclear leukocytes in essential hypertension. These cells could be involved in the pathogenesis of organ injury. Thirty subjects (14 men and 16 women) with essential hypertension were enrolled. In these subjects we determined, at baseline and after in vitro activation with 4-phorbol 12-myristate 13-acetate and N-formyl-methionyl-leucyl-phenylalanine, the polymorphonuclear leukocyte membrane fluidity, obtained by labeling the cells with 1-[4-(trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene, cytosolic Ca2+ concentration, obtained by marking the cells with Fura 2-AM, and integrin pattern (CD11a, CD11b, CD11c, and CD18), by using the indirect immunofluorescence with a flow cytometer. At baseline there was no difference in membrane fluidity between normal subjects and hypertensives, whereas hypertensives showed an increase in cytosolic Ca2+ content and an increase of the phenotypical expression of CD11a, CD11b, and CD18. In normal subjects and in hypertensives, after activation, no variation was found in membrane fluidity and cytosolic Ca2+ content. In normal subjects, after activation, we observed a significant increase of the expression of all adhesion molecules, whereas in hypertensives we found an increase of the expression of CD11b, CD11c, and CD18 but also a decrease of CD11a. The behavior of the polymorphonuclear leukocyte integrin profile may have several explanations, and in particular, the trend of CD11a after chemotactic activation may be related to its cleavage or to an altered integrin phosphorylation/dephosphorylation balance hypothetically present in this clinical condition.

Residual vein thrombosis and onset of post-thrombotic syndrome: Influence of the 4G/5G polymorphism of plasminogen activator inhibitor-1 gene

BACKGROUND Plasminogen activator inhibitor-1 (PAI-1) is the most important inhibitor of plasminogen activator. The functional 4G/5G polymorphism of the gene coding for PAI-1 may affect PAI-1 plasmatic activity, influencing the imbalance between coagulation and fibrinolysis cascades. In this prospective cohort analytic study, we investigated the role of this single nucleotide polymorphism in the persistence of thrombotic lesion and the occurrence of post-thrombotic syndrome. PATIENTS/METHODS In a group of 168 patients with post-surgical deep vein thrombosis of the legs, we analyzed the 4G/5G polymorphism in the promoter of PAI-1 gene and plasmatic PAI-1 activity. Enrolled patients were divided in two groups: patients with 4G/5G polymorphism and increased PAI-1 activity (n=85) and patients without 4G/5G polymorphism and normal PAI-1 activity (n=83). All patients were treated according to current protocols and re-examined after 3, 12 and 36 months in order to evaluate the persistence of thrombotic lesion and the occurrence of post-thrombotic syndrome. RESULTS We found a significantly increased PAI activity in carrier of the 4G allele, who experienced much more frequently a persistence of thrombosis after 3, 12 and 36 months and/or the development of post-thrombosis syndrome, in spite of the anticoagulant treatment. CONCLUSIONS These data not only confirm the role played by PAI-1 activity and by the 4G/5G SNP of the PAI-1 gene, but also suggest that current therapeutic protocols, recommending the administration of low weight molecular heparin and oral anticoagulant for the treatment of deep vein thrombosis, could be non sufficient for patients genetically predisposed to a less efficient clot lysis.

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 patients with macrovascular complications compared with normal subjects (P < 0.001). Platelet membrane C:PL ratio and cytosolic Ca2+ content do not discriminate normal subjects from diabetic patients, and for individual phospholipids, only phosphatidylethanol-amine is decreased in diabetic patients compared with control subjects (P = 0.051). In normal subjects, the polarization degree of TMA-DPH is related to phosphatidylserine (P < 0.05) and phosphatidylcholine (P < 0.05), and in diabetic patients the polarization degree of TMA-DPH is related to C:PL ratio (P < 0.05) and sphyngomyelin (P < 0.05). CONCLUSIONS In type II diabetic patients with macrovascular complications, we observed an abnormality of platelet membrane fluidity, which may contribute to platelet functional alteration present in this clinical condition.

Platelet membrane fluidity and platelet membrane lipid pattern in essential hypertension

In a group of subjects with essential hypertension platelets were studied in resting conditions: platelet membrane fluidity was measured with the fluorescent probe 1.4-(trimethylamino)-phenyl-4-phenylhexatriene (TMA-DPH), platelet membrane cholesterol/phospholipid ratio was evaluated separating the membrane lipids with column chromatography, and platelet membrane individual phospholipids were determined using two-dimensional thin-layer chromatography. From the obtained results, it is evident that platelet membrane fluidity does not differentiate normals from hypertensives; platelet membrane cholesterol/phospholipid ratio is increased in hypertensives, while of the platelet membrane individual phospholipids, only the phosphatidylcholine is increased. In normals and hypertensives, no relation is evident between platelet membrane fluidity, platelet membrane lipid pattern, and systolic and diastolic blood pressure values.

Evaluation of nitric oxide metabolites in a group of subjects with metabolic syndrome

AIM To evaluate the concentration of metabolites (NO(2)(-), NO(3)(-)) of nitric oxide (NO) in metabolic syndrome (MS). MATERIALS AND METHODS We enrolled 106 subjects (45 women and 61 men) with MS of which 43 (14 women and 27 men) with diabetes mellitus and 63 (31 women and 32 men) without diabetes mellitus, and 54 subjects (19 women and 35 men) as control group. The nitric oxide metabolites (nitrite+nitrate=NOx) were evaluated employing the Griess reagent. RESULTS In the whole group of MS subjects was evident, in comparison with control group, a significant increase in NOx. The same finding was also present between control group and diabetic subjects with MS and between control group and nondiabetic subjects with MS. No difference was observed between the two subgroups (diabetic and nondiabetic subjects with MS) about NOx. Contrasting information were obtained examining the linear regression among NOx, age, anthropometric profile, blood pressure values and glycometabolic pattern of subjects with MS. CONCLUSIONS In MS subjects we found a significant increase in NOx not influenced by diabetes mellitus. The NOx is a parameter that must be considered in MS keeping in mind that its behavior is related to chronic inflammation that accompanies this clinical condition.

Lipid Peroxidation, Nitric Oxide Metabolites, and Their Ratio in a Group of Subjects with Metabolic Syndrome

Our aim was to evaluate lipid peroxidation, expressed as thiobarbituric acid-reactive substances (TBARS), nitric oxide metabolites (nitrite + nitrate) expressed as NOx, and TBARS/NOx ratio in a group of subjects with metabolic syndrome (MS). In this regard we enrolled 106 subjects with MS defined according to the IDF criteria, subsequently subdivided into diabetic (DMS) and nondiabetic (NDMS) and also into subjects with a low triglycerides/HDL-cholesterol (TG/HDL-C) index or with a high TG/HDL-C index. In the entire group and in the four subgroups of MS subjects we found an increase in TBARS and NOx levels and a decrease in TBARS/NOx ratio in comparison with normal controls. Regarding all these parameters no statistical difference between DMS and NDMS was evident, but a significant increase in NOx was present in subjects with a high TG/HDL-C index in comparison with those with a low index. In MS subjects we also found a negative correlation between TBARS/NOx ratio and TG/HDL-C index. Considering the hyperactivity of the inducible NO synthase in MS, these data confirm the altered redox and inflammatory status that characterizes the MS and suggest a link between lipid peroxidation, inflammation, and insulin resistance, evaluated as TG/HDL-C index.

Protein oxidation in a group of subjects with metabolic syndrome.

AIMS To examine the protein oxidation, marker of the oxidative stress, in metabolic syndrome (MS). METHODS We enrolled 106 subjects (45 women and 61 men) with MS of which 43 (14 women and 27 men) were with diabetes mellitus and 63 (31 women and 32 men) were without diabetes mellitus, and 54 subjects (19 women and 35 men) as control group. The protein oxidation, expressed as carbonyl groups, was measured by an enzyme-like immunosorbent assay (ELISA) kit (BioCell PC test kit, Enzo Life Sciences AG, Switzerland). RESULTS In the whole group of MS subjects, in comparison with control group, a significant increase in carbonyl groups was present. The same datum was also evident between control group and diabetic subjects with MS and between control group and nondiabetic subjects with MS. No difference was observed between the two subgroups (diabetic and nondiabetic subjects with MS) about NOx. Few information were obtained examining the linear regression among carbonyl groups, age, BMI, waist circumference, blood pressure values and metabolic pattern of MS subjects. CONCLUSIONS In MS subject we observed an increase of protein oxidation not influenced by diabetes mellitus. Several strategies may be employed to reduce this parameter.