Multifractal characterization of morphology of human red blood cells membrane skeleton.

The purpose of this paper is to show applicability of multifractal analysis in investigations of the morphological changes of ultra-structures of red blood cells (RBCs) membrane skeleton measured using atomic force microscopy (AFM). Human RBCs obtained from healthy and hypertensive donors as well as healthy erythrocytes irradiated with neutrons (45 µGy) were studied. The membrane skeleton of the cells was imaged using AFM in a contact mode. Morphological characterization of the three-dimensional RBC surfaces was realized by a multifractal method. The nanometre scale study of human RBCs surface morphology revealed a multifractal geometry. The generalized dimensions Dq and the singularity spectrum f(α) provided quantitative values that characterize the local scale properties of their membrane skeleton organization. Surface characterization was made using areal ISO 25178-2: 2012 topography parameters in combination with AFM topography measurement. The surface structure of human RBCs is complex with hierarchical substructures resulting from the organization of the erythrocyte membrane skeleton. The analysed AFM images confirm a multifractal nature of the surface that could be useful in histology to quantify human RBC architectural changes associated with different disease states. In case of very precise measurements when the red cell surface is not wrinkled even very fine differences can be uncovered as was shown for the erythrocytes treated with a very low dose of ionizing radiation.

Rheological properties of erythrocytes in patients with high risk of cardiovascular disease.

Rheological properties of erythrocytes from patients with high risk of cardiovascular disease (CVD) were analyzed in relation to individual patient risk factors as well as to the medication. Additionally, comparative statistical analysis was performed considering plasma concentration of the selected mediators of vascular endothelium: 6-keto-prostaglandin F(1alpha) (PGF(1alpha)), sVCAM-1 and E-selectin adhesion molecules and interleukin-6 (IL-6). It was found that antihypertensive therapy with angiotensin-converting enzyme inhibitor (ACEI) is accompanied by improvement of RBC rheology: the increase of deformability and the decrease of aggregability. This improvement is probably mediated by endothelial prostacyclin and nitric oxide which are generated by ACEI. A correlation was observed between RBC deformability/aggregability and the patient's hematocrit level, what implicates that the hematocrit level should be explicitly taken into consideration when investigating rheological properties of erythrocytes. A strong relationship was also found between the plasma concentration of sVCAM-1 and patient's age.

Quantitative hopanoid analysis enables robust pattern detection and comparison between laboratories.

Hopanoids are steroid-like lipids from the isoprenoid family that are produced primarily by bacteria. Hopanes, molecular fossils of hopanoids, offer the potential to provide insight into environmental transitions on the early Earth, if their sources and biological functions can be constrained. Semiquantitative methods for mass spectrometric analysis of hopanoids from cultures and environmental samples have been developed in the last two decades. However, the structural diversity of hopanoids, and possible variability in their ionization efficiencies on different instruments, have thus far precluded robust quantification and hindered comparison of results between laboratories. These ionization inconsistencies give rise to the need to calibrate individual instruments with purified hopanoids to reliably quantify hopanoids. Here, we present new approaches to obtain both purified and synthetic quantification standards. We optimized 2-methylhopanoid production in Rhodopseudomonas palustris TIE-1 and purified 2Me-diplopterol, 2Me-bacteriohopanetetrol (2Me-BHT), and their unmethylated species (diplopterol and BHT). We found that 2-methylation decreases the signal intensity of diplopterol between 2 and 34% depending on the instrument used to detect it, but decreases the BHT signal less than 5%. In addition, 2Me-diplopterol produces 10× higher ion counts than equivalent quantities of 2Me-BHT. Similar deviations were also observed using a flame ionization detector for signal quantification in GC. In LC-MS, however, 2Me-BHT produces 11× higher ion counts than 2Me-diplopterol but only 1.2× higher ion counts than the sterol standard pregnane acetate. To further improve quantification, we synthesized tetradeuterated (D4) diplopterol, a precursor for a variety of hopanoids. LC-MS analysis on a mixture of (D4)-diplopterol and phospholipids showed that under the influence of co-eluted phospholipids, the D4-diplopterol internal standard quantifies diplopterol more accurately than external diplopterol standards. These new quantitative approaches permit meaningful comparisons between studies, allowing more accurate hopanoid pattern detection in both laboratory and environmental samples.

[Incidence of leukemia in Krakow in the period 1980-1990]. / Zachorowalnosc na bialaczki w Krakowie w latach 1980-1990.

Based on compiled epidemiological data concerning registered leukemia morbidity in Cracow during the period 1980-1990 a concise calculation of leukemia incidence was made taking into consideration, a year-to-year breakdown of data, leukemia type, sex and age. Our studies showed a rapid increase in total leukemia morbidity in the years 1983-1990 in comparison to 1980-1982. It seems that this increase is caused mainly by the change in leukemia incidence for men.

Evolution of rheological properties of erythrocytes and left ventricular geometry in cardiovascular disease risk patients.

The evolution of rheological properties of erythrocytes and geometrical parameters of left ventricle during therapies aimed at reducing cardiovascular disease (CVD) risk has been investigated. The study group consisted of 29 individuals who were diagnosed with the presence of at least one CVD risk factor at the time of entry to the study. Appropriate therapies were applied and the patients were followed for two years. Two groups of patients could be distinguished. The first group consisted of 12 individuals who were rigorously applying the therapy and for whom blood pressure, total cholesterol, LDL and glucose returned to normal levels. The second group included 17 patients for whom the above mentioned parameters remained pathological in spite of the applied therapy. In the first group, erythrocyte deformability as well as LVMI improved: deformability increased on average by 17% (p < 0.025), whereas LVMI decreased by 8% but not in a statistically significant manner (p < 0.27). In the second group, the results indicate worsening of both hemorheological properties and left ventricular geometry: RBC deformability became lower by 15% (p < 0.00001) and LVMI increased by 18% although this change was not statistically significant (p < 0.19). The results indicate that blood rheology improves when the CVD risk is reduced by administered therapy and worsens when the risk increases. Similar behavior shows LVMI. It is very likely that left ventricular geometry is influenced by blood rheology.

Left ventricular geometry and rheological properties of erythrocytes in patients at cardiovascular disease risk.

The relationship between erythrocyte deformability and aggregability with left ventricular mass index has been examined in patients diagnosed with at least one cardiovascular risk factor but without ongoing coronary heart disease. The group consisted of 66 individuals, 30 men and 36 women, of the average age 57.7 years. For each patient, deformability and aggregability of red blood cells (RBCs) as well as end-diastolic left ventricle diameter (LVD), interventricular septum thickness (IVST) and posterior wall thickness (PWT) were measured. On the basis of the echocardiographical parameters and anthropometric data, left ventricular mass index (LVMI) was calculated. The analysis revealed statistically significant correlation between the LVMI and erythrocyte deformability and aggregability: the LVMI increases with decreasing deformability and is higher in patients with higher aggregability. This finding indicates that the worsening of RBC rheological properties is one of the main factors contributing to alterations of cardiac geometry through the increase of peripheral resistance which, in turn, significantly augments the heart afterload. Given that left ventricular hypertrophy (LVH) is a predictor of cardiovascular morbidity and mortality, the association between hemorheological parameters and left ventricular geometry may be important in clinical practice.