Polymorphonuclear phenotypical expression of CD18, at baseline and after in vitro activation, in several clinical disorders: Revision of our case series.

BACKGROUND: In relation to the different and important roles of the beta2 integrins, we have revisited the expression of polymorphonuclear leukocyte CD18 in several clinical disorders, at baseline and after in vitro activation. SUBJECTS: we have examined subjects with type 1 diabetes mellitus, vascular atherosclerotic disease, type 2 diabetes mellitus without and with macrovascular complications, chronic renal failure on conservative treatment, essential hypertension, deep venous thrombosis, acute ischemic stroke and subjects with venous leg ulcers. METHODS: unfractioned leukocyte suspension was prepared according to the Mikita's method, while the leukocyte were separated into mononuclear and polymorphonuclear cells with a Ficoll-Hypaque medium. Using specific monoclonal antibody, the CD18 expression was evaluated with cytofluorimetric analysis, using FACScan (Becton Dickinson) be Cellquest software; the activation in vitro with PMA was effected according to modified Yasui and Masuda methods. RESULTS: in type 1 diabetes mellitus, at baseline CD18 is under expressed in comparison with normal control, and not changes after PMA activation were observed; in subjects with vascular atherosclerotic disease, in type 2 diabetes mellitus CD18 is over expressed at baseline but does not vary after activation; in subjects with chronic renal failure, essential hypertension and in subjects with acute ischemic stroke the CD18 up-regulate at baseline compared to normal control, and it increases further after activation; in subjects with deep venous thrombosis the CD18 expression is not different from control group at baseline, but it increases after activation; finally, in subjects with venous leg ulcers the CD18 is normally expressed at baseline, and it does not change after PMA activation. CONCLUSIONS: in the different clinical disorders, the trend of this integrin subunit provides some specific information, useful to select the best therapeutic strategy in clinical practice.

Arterial Hypertension and the Hidden Disease of the Eye: Diagnostic Tools and Therapeutic Strategies.

Hypertension is a major cardiovascular risk factor that is responsible for a heavy burden of morbidity and mortality worldwide. A critical aspect of cardiovascular risk estimation in hypertensive patients depends on the assessment of hypertension-mediated organ damage (HMOD), namely the generalized structural and functional changes in major organs induced by persistently elevated blood pressure values. The vasculature of the eye shares several common structural, functional, and embryological features with that of the heart, brain, and kidney. Since retinal microcirculation offers the unique advantage of being directly accessible to non-invasive and relatively simple investigation tools, there has been considerable interest in the development and modernization of techniques that allow the assessment of the retinal vessels' structural and functional features in health and disease. With the advent of artificial intelligence and the application of sophisticated physics technologies to human sciences, consistent steps forward have been made in the study of the ocular fundus as a privileged site for diagnostic and prognostic assessment of diverse disease conditions. In this narrative review, we will recapitulate the main ocular imaging techniques that are currently relevant from a clinical and/or research standpoint, with reference to their pathophysiological basis and their possible diagnostic and prognostic relevance. A possible non pharmacological approach to prevent the onset and progression of retinopathy in the presence of hypertension and related cardiovascular risk factors and diseases will also be discussed.

MIF rs755622 and IL6 rs1800795 Are Implied in Genetic Susceptibility to End-Stage Renal Disease (ESRD).

Chronic kidney disease (CKD) is characterized by an increased risk of kidney failure and end-stage renal disease (ESRD). Aging and comorbidities as cardiovascular diseases, metabolic disorders, infectious diseases, or tumors, might increase the risk of dialysis. In addition, genetic susceptibility factors might modulate kidney damage evolution. We have analyzed, in a group of ESRD patients and matched controls, a set of SNPs of genes (Klotho rs577912, rs564481, rs9536314; FGF23 rs7955866; IGF1 rs35767; TNFA rs1800629; IL6 rs1800795; MIF rs755622, rs1007888) chosen in relation to their possible involvement with renal disease and concomitant pathologies. Analysis of the raw data did indicate that IL6 rs180795 and MIF rs755622 SNPs might be markers of genetic susceptibility to ESRD. In particular, the C positive genotypes of MIF rs755622, (dominant model) seem to be an independent risk factor for ESDR patients (data adjusted for age, gender, and associated pathologies). Stratifying results according to age MIF rs755622 C positive genotype frequencies are increased in both the two age classes considered (<59 and ≥59-year-old subjects). Analyses of data according to gender allowed us to observe that ESRD women shoved a significantly reduced frequency of genotypes bearing IL6 rs180795 C allele. In addition, MIF rs755622 might interact with diabetes or hypercholesterolemia in increasing susceptibility to ESRD. In conclusion, our data indicate that some polymorphisms involved in the regulation of both renal function and inflammatory response can influence the evolution of chronic kidney disease and suggest that the modulation of the activities of these and other genes should also be considered as therapeutic targets on to intervene with innovative therapies.

The function of matrix metalloproteinase-9 (MMP-9) and its tissue inhibitor (TIMP-1) in several clinical conditions: Results and analysis of our survey.

The goal of this research was to evaluate the plasma concentration of MMP-9 and its tissue inhibitor (TIMP-1) in different clinical conditions. It included several groups of subjects: 31 overweight subjects; 91 obese adults divided into two subgroups according to the BMI value (BMI 30-35 Kg/m2 and BMI > 35 Kg/m2); 90 subjects with metabolic syndrome (MS) divided into two subgroups (with and without diabetes mellitus); 100 subjects with preclinical carotid atherosclerosis (PCA) divided according to the number of cardiovascular risk factors and to the insulin resistance degree; 48 subjects with obstructive sleep apnoea syndrome (OSAS) divided according to the apnoea/hypopnea index (AHI); 27 subjects with chronic kidney disease (CKD) on conservative management; 31 subjects with CKD on regular haemodialysis treatment. We have found a significant increase of MMP-9 and TIMP-1 in overweight subjects, in obese adult and in MS subjects. In obese adults, the behaviour of these two parameters was not influenced by the degree of obesity, while in the group of MS subjects both these parameters were clearly influenced by the presence of diabetes mellitus. In subjects with PCA, we observed an increase of MMP-9 associated with a significant decrease of TIMP-1; the same trend was found by subdividing the entire group in accordance with the number of cardiovascular risk factors and with the insulin resistance degree. In subjects with OSAS, we noted an increase in MMP-9 and TIMP-1; this increase was more evident in subjects with OSAS having AHI > 30. In individuals with CKD on conservative and haemodialysis treatment we have found, at baseline, a marked increase in MMP-9 and a significant decrease of TIMP-1. In dialyzed subjects, after a standard dialysis session was noted, a significant increase in MMP-9 was associated with a further decrease in TIMP-1.

Relationship of choroidal thickness with pulsatile hemodynamics in essential hypertensive patients.

Controversy exists about the association of choroidal thickness (CTh) with blood pressure (BP) values. There is some evidence suggesting that central hemodynamics changes are associated with microvascular disease. Our study was aimed to assess the relationships between CTh and clinic and 24-h BP and between CTh and estimated 24-h aortic pulse pressure (aPP), 24-h aortic systolic BP (aSBP), and 24-h aortic augmentation index (aAIx) in a group of hypertensive patients. We enrolled 158 hypertensive subjects (mean age 48 ± 13 years) all of which underwent evaluation of the choroidal district by Swept-Source optical coherence tomography (SS-OCT) and 24-h BP monitoring, in order to measure peripheral BP and to estimate central hemodynamic parameters. Inverse significant correlations of clinic PP, 24-h aPP, 24-h aSBP, and 24-h aAIx with thicknesses of central ring, inner ring, and outer ring of the choroid and its overall average were found. The strongest of these correlations was that relating 24-h aPP with overall average choroidal thickness (r = -.531; P < .001). When we divided the study population in subjects with 24-h aPP above and below the median value (35 mm Hg), CTh were thinner in subjects with higher values of 24-aPP as compared to those with lower ones, even after adjustment for age, and other potential confounders. The relationships of CTh with 24-h aPP remained significant also taking into account the effects of various covariates in linear multiple regression analyses. Our findings support the concept of a cross-talk between macro- and microcirculation.

Behaviour of carbonyl groups in several clinical conditions: Analysis of our survey.

Protein carbonylation is a marker of oxidative protein damage, that is likely involved in the pathogenesis of several diseases. The aim of this study was to evaluate the protein carbonyl (PC) groups in different clinical conditions. It included different groups of subjects: 81 trained subjects; 23 subjects with mild essential hypertension; 31 middle-aged subjects with metabolic syndrome (MS); 106 subjects with MS not selected for age (subdivided into two subgroups, with and without diabetes mellitus); 91 obese adults subdivided in two subgroups (BMI 30-35 Kg/m2 and BMI > 35 kg/m2); 48 subjects with obstructive sleep apnea syndrome (OSAS) subdivided in accordance with the apnea/hypopnea index (AHI); 27 subjects with chronic kidney disease (CKD) on conservative therapy; 31 subjects with CKD on haemodialysis treatment; and 50 subjects with juvenile myocardial infarction. PC groups were reduced in trained subjects in comparison with sedentary controls, while no variation was observed in mild essential hypertension. PC groups were increased in MS subjects and in adult obese subjects. In MS subjects the PC groups were not influenced by the presence of diabetes mellitus and in adult obese subjects were not influenced by the obesity degree. In OSAS subjects only those with AHI > 30 showed an increase of PC groups. PC groups increased in CKD subjects undergoing conservative treatment and haemodialysis therapy. In dialyzed subjects, after a standard dialysis session, there was a marked increase in PC groups. In juvenile myocardial infarction PC groups were higher than in controls; there was no difference between STEMI and NSTEMI and their concentration was unaffected by the number of cardiovascular risk factors or stenosed coronary vessels.

[Analysis of the parameters, traditional or not, for the evaluation of the metabolic acidosis]. / Considerazioni sui parametri di valutazione, tradizionali e non, delle acidosi metaboliche.

Despite huge progress in acidbase knowledge, several confusing, irrational and controversial issues still remain. Acid-base disturbances have been usually evaluated with the traditional Henderson-Hasselbach method and with BE evaluation that seem inadequate since they define the magnitude of metabolic acidosis rather than its cause. Some studies have shown that the traditional approach is often not able to highlight the complicated acid-base disorders in critically ill patients; in these subjects, the possibility to identify tissue acids could offer a greater prognostic value than the evaluation of traditional parameters. An alternative approach is the Stewarts physiochemical method that defines the aetiology of a metabolic acidosis by quantifying the tissue acids. But the clinical utility of this method is limited due to its mathematical complexity. Therefore, some parameters of simplification were proposed in order to allow greater clinical applicability of this system. Specifically, it was observed that in the presence of metabolic acidosis, the chloride/sodium ratio (Cl-/Na+ ratio) or the sodium-chloride difference (DiffNa-Cl) would be useful indicators of the presence of unmeasured anions (UMA) and/or lactate.

Protein oxidation in chronic kidney disease.

An imbalance between oxidative processes and antioxidant systems has been widely demonstrated in chronic kidney diseases (CKD). In this study we enrolled 26 healthy subjects, 27 patients with CKD on conservative treatment (CT-CKD) with various degrees of renal failure, and 31 CKD subjects in haemodialysis treatment (HD-CKD), evaluated before and after a standard haemodialysis session. In each group we measured protein carbonyl groups (PC) as an index of protein oxidation, lipid peroxidation (TBARS) and two plasma markers of leukocyte activation, elastase and myeloperoxidase (MPO). In CT-CKD subjects the PC level was significantly higher than in normal controls, and it was negatively correlated with creatinine clearance. In HD-CKD patients the PC concentration was significantly increased also in comparison with CT-CKD. An increase in TBARS was present both in CT-CKD and in HD-CKD patients, but in HD-CKD patients TBARS were lower than in CT-CKD. Elastase was increased in both CKD groups, while MPO was not different among control and patient groups. In HD-CKD patients the HD session was followed by a further increase in PC, as well as by an increase in elastase and MPO, whereas TBARS did not change. Protein oxidation accelerates the glycation processes and seems to be connected with the chronic inflammatory state detectable in renal failure, although we did not observe any significant correlation between PC level and leukocyte activation markers.

Elastase, myeloperoxidase, nitric oxide metabolites and oxidative status in subjects with clinical stable chronic renal failure on conservative treatment.

We evaluated, in a group of 41 CRF undialyzed subjects (29 men and 12 women, mean age 64.1 +/- 11.3 years), some parameters that reflect leukocyte activation (elastase, myeloperoxidase - MPO), plasma NO metabolites (NO(x)) and the oxidative status (lipid peroxidation expressed as thiobarbituric acid-reactive substances (TBARS) and total antioxidant status (TAS). Elastase was determined, on plasma separated from fasting venous blood, as elastase/alpha1-proteinase inhibitor complex. MPO was evaluated employing the Myeloperoxidase ELISA kit. The NO production was evaluated by a micromethod. The oxidation of polyunsaturated fatty acids was evaluated in plasma by detection of the thiobarbituric acid-reactive substances (TBARS). Total antioxidant status was measured by spectrophotometry. We found a significant increase of elastase, TBARS and NO(x), without any significant variation of MPO and TAS. In this group of CRF subjects, no statistical correlation was found between these examined parameters, creatinine level, creatinine clearance, leukocyte count and hemoglobin level. These findings need to be underlined if we consider that chronic renal failure is an inflammatory condition and this research furtherly supports literature data regarding the role of activated leukocytes in the development of the vascular complications. These observations explain why the examination of leukocyte count and function could become a tool to verify the clinical outcome in these patients.

Nitric oxide metabolites (nitrite and nitrate) in young patients with recent acute myocardial infarction.

Nitric oxide (NO) has a role in the pathophysiology of acute and chronic cardiovascular events. We studied the plasma concentration of NO stable end products (nitrite and nitrate--NOx) in 43 patients aged<46 years, with recent acute myocardial infarction (AMI). The evaluation was effected at the initial stage, after 3 and 12 months. We subdivided the patients into subgroups according to the number of the main cardiovascular risk factors and to the extent of coronary disease. In the whole group the NOx concentration was initially increased and progressively decreased after 3 and 12 months, remaining at both times significantly higher than in control subjects. The patients with more risk factors had a significantly higher NOx concentration. In conclusion, the persisting high NOx concentration in AMI patients is the expression of a prolonged inflammatory condition and is significantly influenced by the simultaneous presence of several cardiovascular risk factors.

Lipid peroxidation and total antioxidant status in juvenile myocardial infarction.

In a group of young subjects with acute myocardial infarction (AMI) (97 men and 8 women; mean age 39.6+/-5.5 years) we examined the thiobarbituric acid - reactive substances and the total antioxidant status at the initial stage and after 12 months. The same parameters were examined in a group of 55 control subjects. Our results show that, while in control subjects there was a negative correlation (p<0.001) between these two parameters, no correlation was found in juvenile myocardial infarction at the initial stage as well as after 12 months.

Wine consumption and renal diseases: new perspectives.

Investigations into the relation between wine consumption and kidney disease have been limited. Patients with chronic renal failure show accelerated atherosclerotic damage and, considering the well-known protective effect of wine on the cardiovascular system, moderate wine consumption might be advantageous. Oxidative stress and endothelial dysfunction, which are inter-related, play a role in the pathophysiology of many renal diseases, including acute and chronic renal failure. Ethanol and non-alcoholic wine components, especially polyphenols, influence oxidative balance and endothelial function. Although long-term alcohol abuse has been associated with many renal alterations in humans, in experimental studies wine polyphenols enhanced kidney antioxidant defenses, exerted protective effects against renal ischemia/reperfusion injury, and inhibited apoptosis of mesangial cells. Moreover, in diabetic patients the administration of moderate amounts of red wine and a polyphenol-enriched diet slowed the progression of diabetic nephropathy. Moreover, the unfavorable effect of ethanol on blood pressure control seems to be counterbalanced by polyphenol protective effects. There is convincing evidence of a beneficial effect of controlled wine consumption patients with renal disease, but controlled clinical trials are needed to confirm this hypothesis.

Wine, diet, and arterial hypertension.

Hypertension is one of the leading causes of death in developed countries, and the number of prehypertensive patients is increasing. The beneficial effects of moderate wine consumption on cardiovascular diseases have been demonstrated, along with the healthy influence of a Mediterranean dietary pattern. The association of these 2 factors on hypertension and its complications is considered here. As wine polyphenols exert a vasorelaxing action, they might positively influence the hemodynamic situation of these patients. These effects could be enhanced by dietary constituents, such as garlic, onions, and olive oil, which are widely employed in Mediterranean cooking. By evaluating many studies performed in animal models and in humans, the authors conclude that moderate wine consumption, if associated with a healthy dietary pattern, such as the Mediterranean one, could help hypertensive patients to ameliorate their arterial pressure and quality of life by reducing cardiovascular morbidity and mortality rates.

Polymorphonuclear leukocyte membrane fluidity and cytosolic Ca2+ concentration in subjects with vascular atherosclerotic disease subdivided according to its extent.

An abnormal activation state of polymorphonuclear leukocytes (PMN) plays a key role in organ injury induced by vascular atherosclerotic disease (VAD) and diabetes mellitus (DM). PMN membrane fluidity and cytosolic Ca2+ content can be considered markers of PMN activation. In this research we evaluated the PMN membrane fluidity and cytosolic Ca2+ content in VAD subjects with and without type 2 DM and examined the association between these parameters and the mono- or polyvascular localization. We enrolled 155 VAD subjects, including 92 non-diabetic (group A: mean age 63.6 +/- 9.2 years) and 63 diabetic patients (group B: mean age 65.4 +/- 7.8 years). Among group A 63 patients had monovascular and 29 polyvascular disease; among group B 30 patients had monovascular and 22 polyvascular disease. In each patient we evaluated the PMN membrane fluidity labelling the cells with the fluorescent probe 1,4-(trimethylamino)-phenyl-4-phenylhexatriene (TMA-DPH) and the PMN cytosolic Ca2+ content marking the cells with the fluorescent probe Fura 2-AM. PMN membrane fluidity did not discriminate normal subjects from diabetic and non-diabetic VAD subjects, while cytosolic Ca2+ content was increased in both groups. PMN membrane fluidity did not distinguish normal subjects from mono- or polyvascular VAD patients with and without type 2 DM. PMN cytosolic Ca2+ content was increased especially in monovascular VAD patients; both mono- and polyvascular VAD subjects with DM had a PMN cytosolic Ca2+ content higher than normals. Our results show the presence of an increased PMN cytosolic Ca2+ content in diabetic and non-diabetic VAD subjects but no association was observed between this increase and the mono- or polyvascular localization.

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 CD18 in CRF patients. In normal subjects, after activation with both agents, we noted an increase of all adhesion molecules, while in CRF patients we found an increase in the expression of CD11b, CD11c and CD18 but not of CD11a. The altered behaviour of the PMN integrin pattern in mild CRF patients, likely reflecting a state of PMN activation, might have a pathophysiological significance, considering the high incidence of cardiovascular events in CRF.

Polymorphonuclear leukocyte integrin profile in vascular atherosclerotic disease.

Leukocyte-endothelial interactions could have a pathogenic role in atherogenesis. Adhesion molecules expressed by endothelial cells, such as intercellular adhesion molecule 1 (ICAM-1), interact with leukocyte integrins mediating the firm adhesion of leukocytes to endothelium which is followed by their transendothelial migration. The aim of our research was to evaluate polymorphonuclear leukocyte (PMN) integrin expression, at baseline and after activation, in a group of subjects with chronic vascular atherosclerotic disease (VAD). In 27 subjects with VAD we examined, at baseline and after in vitro activation with 4-phorbol 12-myristate 13-acetate (PMA), the PMN integrin pattern (CD11a, CD11b, CD11c, CD18) using indirect immunofluorescence and a flow cytometer. At baseline VAD subjects showed an increase of CD11a and CD18 and a decrease of Cd11b and Cd11c as compared to normal subjects. After activation, in normal subjects, we found an increase in the expression of all integrins, while in VAD subjects we observed an increase of CD11b and Cd11c and a decrease of Cd11a and CD18. In VAD subjects, at baseline, the upregulation of Cd11a and CD18 may reflect PMN in vivo activation; after in vitro activation, the decrease of CD11a may be related to the lack of cytoplasmic deposits of this molecule, while CD18 might be internalized. The integrin behaviour pattern in chronic VAD deserves further investigation, considering that integrins are potential targets of therapeutical strategies, with the aim of preventing the atherosclerotic plaque progression and acute ischaemic events.