Association of sulfur content in erythrocytes with cardiovascular parameters and blood pressure.

OBJECTIVE: The study aims at assessing the relationship between blood pressure, heart geometry parameters, and the erythrocyte content of sulfur, potassium, chlorine and phosphorus, in a group of patients with ambulatory systolic and diastolic blood pressure (SBP, DBP) below 140 or 90 mm Hg, respectively, who were otherwise healthy and untreated. METHODS: The study group consisted of 42 adults recruited in a primary care setting. The individuals were healthy, not undergoing any therapy and free from smoking. For each individual, data were obtained on: average 24-hour SBP and DBP, left ventricle geometry, complete blood count, lipids profile, fibrinogen, hs-CRP and the erythrocyte concentration of sulfur (S), potassium (K), chlorine (Cl) and phosphorus (P). RESULTS: Multivariate regression analysis showed statistically significant relationships of diastolic posterior wall thickness (PWTd) and relative wall thickness (RWT) with the concentration ratio of sulfur and potassium (S/K) in erythrocytes: PWTd and RWT increase as the S/K ratio increases. Also, SBP was found to be positively correlated with the S/K ratio. CONCLUSIONS: The increase in sulfur content in RBCs could be an indicator of the downregulation of nitric oxide (NO) erythrocyte bioavailability exerted by endogenously produced hydrogen sulfide (H2S), and, in consequence, a marker of the development of hypertension and/or adverse changes in heart geometry.

Longitudinal effects of a nurse-managed comprehensive cardiovascular disease prevention program for hospitalized coronary heart disease patients and primary care high-risk patients.

BACKGROUND: The EUROACTION study (nurse­coordinated multidisciplinary, family­based cardiovascular disease prevention program) documented the efficacy of a nurse­managed, comprehensive prevention program in reducing risk factors for cardiovascular disease (CVD). No information was available on survival. AIMS: The aim of the study was to assess the effects of EUROACTION intervention on CVD risk factors and 12­year survival in the Polish component of the study. METHODS: Two district hospitals and 2 primary care practices were allocated randomly to intervention (INT) or usual care (UC). The primary endpoints were lifestyle and risk factors changes at 1­year follow­up. Differences in survival were analyzed using the multivariable Cox proportional hazards regression models. RESULTS: The study involved 628 patients with coronary heart disease (CHD) and 711 high­risk patients. Compared to UC, INT patients achieved healthier lifestyles and a larger reduction of risk factors at 1 year but these differences were not maintained 12 years after the intervention. Less deaths occurred in patients from the INT hospital and from INT primary practice (hazard ratio [HR], 0.58; 95% CI, 0.42-0.82 and HR, 0.53; 95% CI, 0.3-0.95, respectively). Adjustment for the covariates slightly attenuated the estimates and removed significance (HR, 0.74; 95% CI, 0.52-1.04 and HR, 0.66; 95% CI, 0.36-1.24, respectively). For combined CHD and high­risk patient groups, compared with UC, INT patients had a 36% lower risk of death after adjustment for age, sex, and history of CHD (HR, 0.64; 95% CI, 0.48-0.86). CONCLUSIONS: The impact of the EUROACTION intervention on lifestyle and CVD risk factors could have contributed to lower mortality in INT coronary and high­risk patients. These results emphasize the need for sustaining the interventions to help patients maintain a healthy lifestyle.

Erythrocyte heme­oxygenation status indicated as a risk factor in prehypertension by Raman spectroscopy.

Raman spectroscopy of erythrocytes provides detailed information about the structure and status of heme moiety, which can be used to provide new insights into molecular pathogenesis of several diseases. In this study, we present the first Raman spectroscopy investigations of the effect of hemoglobin oxygenation in the context of hypertensive disease. The experimental data was subjected to Logistic Regression, which indicated heme­oxygenation status as an important risk factor alongside other clinical parameters. The 1605/1621 cm-1 band ratio was selected as an optimal Raman metric for risk assessment and along with other band ratios (1583, 1639, 1310 cm-1) related to heme status and when combined with clinical data via logistic regression gave an Area Under the Curve (AUC) >0.95 for prehypertension risk prediction. The work demonstrates the feasibility of Raman spectroscopy to distinguish between prehypertensive and normotensive states. Simultaneously, it is implied that the etiology of the high blood pressure progression may be connected with the changes in hemoglobin oxygenation.

Triglycerides as indicators of erythrocyte hemoglobin oxygen-binding properties1.

OBJECTIVES: To assess the relationships of clinical and laboratory parameters, with erythrocyte oxygen concentration. METHODS: The study group consisted of 47 healthy adults out of which 16 showed slightly higher blood pressure and were classified as individuals with prehypertension. For each individual, data were obtained on: systolic and diastolic blood pressure (SBP, DBP), blood morphology, lipids profile, fibrinogen, hs-CRP. The erythrocyte oxygen concentration was assessed with the Raman spectroscopy technique. Arithmetic means for all laboratory parameters were estimated by oxygen concentration tertiles and tested for statistical significance of linear trends across tertiles. RESULTS: The multivariate regression analysis showed statistically significant negative relationship of triglycerides level with oxygen concentration. CONCLUSIONS: A hypothesis has been formulated that triglycerides level is an indicator of the erythrocyte ability to transport oxygen to the tissues.

The influence of very small doses of alpha radiation on the stability of erythrocytes.

Our aim was to study the influence of low doses (0.2-4 µGy) of α radiation on the stability of human erythrocytes isolated from healthy and diabetic erythrocytes. Absorption spectroscopy was used to measure the level of red blood cell (RBC) hemolysis, along with Mössbauer spectroscopy, which is a highly specific method suited to monitoring various hemoglobin forms. States of hemoglobin are sensitive to a homeostatic imbalance in red blood cells. Changes in the membrane skeleton organization of irradiated erythrocytes isolated from healthy donors were studied using atomic force microscopy (AFM). Hemolysis, in healthy red blood cells, showed characteristic discontinuities, depending on the α particle flux and the exposure time to the low doses applied. This phenomenon was not observed in severe diabetic cases, which could be a result of modified protein-lipid-sugar complexes and the attenuation/absence of some antioxidative enzymatic processes in their RBC membranes. Similar effects were also observed for red blood cells treated with low doses of neutron and γ-radiation. AFM measurements demonstrated a reorganization of the RBC membrane skeleton network depending on the time of RBC exposure to α radiation. This suggests that the changes in the activity of the acute defense processes against free radicals which are activated within the erythrocyte membrane irradiated with α-particles could additionally be up- or down regulated by modifications to the membrane-skeleton network. However, even the highest dose of α radiation applied in these studies did not cause any significant changes in the ability of hemoglobin to transport oxygen. Microsc. Res. Tech. 80:131-143, 2017. © 2016 Wiley Periodicals, Inc.

Association of red blood cell distribution width, inflammation markers and morphological as well as rheological erythrocyte parameters with target organ damage in hypertension.

OBJECTIVES: To assess the relationships of red blood cell distribution width (RDW), mean corpuscular hemoglobin concentration (MCHC) and erythrocyte deformability with pathological changes of selected target organs, and with inflammation markers interleukin-6 (IL-6) and fibrinogen, in a group of newly diagnosed, never-treated and otherwise healthy hypertensive patients. METHODS: The study group consisted of 101 adults divided into three sub-groups: 37 diagnosed arterial hypertension, 29 with hypercholesterolemia, and 35 healthy. The individuals with hypertension or hypercholesterolemia were otherwise healthy and were not on any therapy prior to entering the study. For each individual, data were obtained on: systolic and diastolic blood pressure (SBP, DBP), pulse wave velocity (PWV), carotid intima media thickness (IMT), left ventricle geometry, blood morphology, lipids profile, fibrinogen, CRP, IL-6 and red blood cell deformability index (DI). RESULTS: In the group of hypertensives, the multivariate regression analysis showed significant relationship of RDW with PWV, IL6 and fibrinogen. Also, RDW was found to be correlated with MCHC and DI, and MCHC was significantly related to IMT and IL-6. CONCLUSIONS: A hypothesis has been formulated that the development of target organ damage in hypertension is accompanied by the increasing impairment of erythropoiesis. This process may be mediated by inflammation.

Erythrocyte membrane properties in patients with essential hypertension.

In spite of the extensive research efforts that have been conducted over the last decades, it is still very difficult to point out genetic determinants or environmental conditions responsible for the development of essential hypertension. We searched for differences in the RBC membrane skeleton structure and O2 membrane permeability between RBCs from patients with both essential arterial hypertension and hypercholesterolemia, from patients having only hypercholesterolemia and from healthy donors. The topography of RBCs and the content of various hemoglobin forms were detected using atomic force microscopy and Mössbauer spectroscopy, respectively. We found that the membrane skeleton of RBCs from healthy donors displayed a well-known honeycomb pattern, whereas in patients with essential hypertension and/or hypercholesterolemia, who had never received anti-hypertensive therapy, it displayed a corncob pattern. Hypertensive RBCs had an oval shape and the average lateral to longitudinal diameter ratio for the changed cells (about 70%) did not exceed 0.80. We observed that after the incubation of RBCs under high nitrogen (low O2) pressure at room temperature and then their transfer into 85 K, a content of oxyHb (deoxyHbOH) already after 1 h reached a stable level of about 85 ± 3% (15 ± 3%) in hypertensives, whereas in healthy individuals it showed a decrease for deoxyHbOH and an increase for oxyHb, which stabilized at a level of about 81 ± 5% and 19 ± 5%, respectively, only after 9 h. Quantitative analysis of the Δ(oxyHb) change estimated as the difference between the oxyHb level measured after 9 and 2 h at 85 K under low N2 pressure (to slow down oxyHb formation) was significantly higher in normotensives than in hypertensive patients with or without hypercholesterolemia, 19.9 versus -4.2, p < 0.02. Our findings indicate an impaired oxygen release by Hb in RBCs of patients with hypertension under low oxygen pressure which if present in vivo may cause hypoxemia and, in turn, further increase of blood pressure.

Depth-sensing analysis of cytoskeleton organization based on AFM data.

Atomic force microscopy is a common technique used to determine the elastic properties of living cells. It furnishes the relative Young's modulus, which is typically determined for indentation depths within the range 300-500 nm. Here, we present the results of depth-sensing analysis of the mechanical properties of living fibroblasts measured under physiological conditions. Distributions of the Young's moduli were obtained for all studied cells and for every cell. The results show that for small indentation depths, histograms of the relative values of the Young's modulus described the regions rich in the network of actin filaments. For large indentation depths, the overall stiffness of a whole cell was obtained, which was accompanied by a decrease of the modulus value. In conclusion, the results enable us to describe the non-homogeneity of the cell cytoskeleton, particularly, its contribution linked to actin filaments located beneath the cell membrane. Preliminary results showing a potential application to improve the detection of cancerous cells, have been presented for melanoma cell lines.

Effects of low doses of gamma rays on the stability of normal and diabetic erythrocytes.

We studied the influence of low doses of γ radiation (from 0.04 to 1.8 mGy) on the stability of human red blood cells (RBC) from healthy donors and diabetic patients using absorption spectroscopy. Because of the alteration of many enzymatic pathways in diabetic RBCs resulting in strong modification of the lipid and protein membrane components one could expect that the ionizing γ-radiation should influence the stability of the healthy and diabetic cells in a different way. Indeed, distinct discontinuities and monotonic changes of hemolysis detected in the healthy and diabetic RBCs suggest that various enzymatic and chemical processes are activated in these membranes by γ radiation. Mössbauer measurements showed that only the highest applied dose of γ radiation caused modification of hemoglobin in both types of RBCs.

Relevance of erythrocyte deformability to the concentration of soluble cell adhesion molecules and glomerular filtration rate in patients with untreated essential hypertension.

Relationship between erythrocyte deformability and: a) soluble cell adhesion molecules concentration, b) glomerular filtration rate (eGFR) has been investigated in three study groups: a group of 20 patients with diagnosed arterial hypertension, a group of 20 individuals with exclusively hypercholesterolemia and a group of 22 healthy persons. The individuals with hypertension or hypercholesterolemia were free of any other cardiovascular disease risk factor and were not on any therapy prior to entering the study. Clinical and laboratory data included systolic and diastolic blood pressure (obtained by ABPM), lipids profile, eGFR, red blood cell (RBC) deformability (assessed by shear stress laser diffractometry) and levels of circulating soluble vascular adhesion molecules-1 (sVCAM-1) as well as soluble intracellular adhesion molecules-1 (sICAM-1). In the group of hypertensives, RBC deformability and concentration of circulating soluble adhesion molecules showed statistically significant negative correlations: RBC deformability decreases with increasing level of: a) sVCAM-1, R = -0.61, p < 0.002, b) sIVCAM-1, R = -0.53, p < 0.009. In parallel, statistically significant increase of eGFR was observed with rising erythrocyte deformability, R = 0.60, p < 0.005. In the groups of healthy individuals and patients with hypercholesterolemia there was no sign of any correlations between the considered parameters. The observed correlations suggest that in patients diagnosed exclusively with hypertension, firstly, erythrocyte deformability may serve as a marker of endothelial dysfunction and, secondly, red blood cells may be mediators of adverse changes in kidneys.

Women with prehypertension in primary care - Risk profile on the basis of selected cardiovascular risk factors.

BACKGROUND: Demographic, social and economic trends will serve to increase the importance of women as healthcare consumers. Design. The aim of the study was to assess cardiovascular (CV) risk in the normotensive female patients during single visit to primary care (PC) offices. METHODS: Demographic data, history of coronary heart disease (CHD), diabetes (DM), smoking habit and family history of CV diseases were obtained from women who visited general practitioners. Moreover, blood pressure (BP), pulse rate, weight and height used to calculation body mass index (BMI) and waist circumference (WC) were performed. Prehypertension was defined as a systolic BP (SBP) of 120-139 mmHg, and/or a diastolic BP (DBP) of 80-89mmHg. RESULTS: Prehypertension was observed in 21.5% of the whole group of female PC patients. SBP, DBP, BMI and WC revealed significant trends towards increase with age among both prehypertensives (p<0.001) and normotensives (SBP, BMI, WC: p<0.001; DBP: p<0.05) and in the whole group (p<0.001). Nevertheless, heart rate (HR) significantly increased with age only among prehypertensive women (p<0.05). The CV risk of the studied adult women increased progressively with presence of overweight, obesity and visceral obesity. The CV risk of the youngest groups was associated mainly with high prevalence of smoking, and with high prevalence of CHD and DM among the oldest female patients. CONCLUSIONS: The prevalence of majority of CV risk factors increase with age among both prehypertensive and normotensive women, which should stimulate PC practitioners to identify and modify them.

Estimation of free copper ion concentrations in blood serum using T(1) relaxation rates.

The water proton relaxation rate constant R(1)=1/T(1) (at 60 MHz) of blood serum is substantially increased by the presence of free Cu2+ ions at concentrations above normal physiological levels. Addition of chelating agents to serum containing paramagnetic Cu2+ nulls this effect. This was demonstrated by looking at the effect of adding a chelating agent-D-penicillamine (D-PEN) to CuSO4 and CuCl2 aqueous solutions as well as to rabbit blood serum. We propose that the measurement of water proton spin-lattice relaxation rate constants before and after chelation may be used as an alternative approach for monitoring the presence of free copper ions in blood serum. This method may be used in the diagnosis of some diseases (leukaemia, liver diseases and particularly Wilson's disease) because, in contrast to conventional methods like spectrophotometry which records the total number of both bound and free ions, the proton relaxation technique is sensitive solely to free paramagnetic ions dissolved in blood serum. The change in R(1) upon chelation was found to be less than 0.06 s(-1) for serum from healthy subjects but greater than 0.06 s(-1) for serum from untreated Wilson's patients.

[Cardiovascular risk factors in the elderly patients in primary care]. / Czynniki ryzyka sercowo-naczyniowego wsród starszych pacjentów podstawowej opieki zdrowotnej.

OBJECTIVE: Aim of the study was to assess cardiovascular risk in the elderly patients during single visit to primary care (PC) offices. MATERIAL AND METHODS: The study was performed in 256 units of PC in 2004 and 2005 year. Demographic data, history of diabetes and smoking habit were obtained from patients older than 60 years old, who visited general practitioners. Moreover, blood pressure, pulse, weight and height used to calculate body mass index (BMI), waist circumference and glucose strip tests were performed. Blood pressure levels were classified according to the guidelines, BMI > or = 30 kg/m2 was considered as obesity, waist > or = 88 cm for women and > or = 102 cm for men were criterions of visceral obesity. Passing glucose > 125 mg/dl (6,9 mmol/l) was noted. Mean values and the performance of studied parameters were compared between three age groups: 61-70, 71-80, and > 80 years and reference to gender. RESULTS: 26 801 patients aged between 61 and 102 years were examined, 59.5% were women, 81.9% were hypertensives, more often men. Isolated systolic hypertension (ISH) was diagnosed in 1/4 of subjects, more often in women. Along with age, the frequency of ISH and severe stages of hypertension increased. 39.8% subjects had visceral obesity, more often women (51.8% vs. 21.9%). Frequency of visceral obesity decreased along with age but constantly was significantly greater among women than men. Diabetes and glucose > 125 mg/dl was stated in 26.0% and 26.3%, respectively. Frequency of smokers was 19.3%, more frequent in men (30.9% vs. 11.5%), and decreased with age. CONCLUSIONS: Increased cardiovascular risk of primary care elderly patients was mainly connected with high frequency of hypertension, especially ISH in the oldest. Among elderly men additional risk resulted from widespread smoking habit, while among women from high frequency of obesity, especially visceral.

Erythrocyte stiffness in diabetes mellitus studied with atomic force microscope.

The mechanical properties of erythrocyte membrane have been studied using an atomic force microscope. Measurements were carried out on blood samples taken from 7 diabetes mellitus patients and 8 healthy individuals. For each blood sample a distribution of a Young's modulus was constructed. It has been found that both the mean value and the width of the distribution in diabetic patients exceed the corresponding results for healthy persons by a factor greater than 3. The high sensitivity of the atomic force microscopy and the ability to measure the full distribution of the erythrocyte membrane Young's modulus makes it a unique, powerful and promising tool in studies of the membrane stiffness of red blood cells.

Erythrocyte stiffness probed using atomic force microscope.

The stiffness of erythrocytes in patients (N=45) suffering from certain disorders, such as coronary disease, hypertension, and diabetes mellitus has been assessed using the Atomic Force Microscopy (AFM) and compared with that in a group of healthy individuals (N=13). For each blood sample, around 20 erythrocytes were selected at random and the stiffness of each one was probed in 20-30 arbitrarily chosen points. From these results, distributions of the cell Young's modulus (YM) were determined. Average values and widths of YM distributions significantly increased in samples taken from diabetes mellitus patients and cigarette smokers, as compared to those taken from healthy donors. At the same time, the average values of YM were found to increase as a function of the patient's age. We demonstrated that the atomic force microscope is a very sensitive tool for determination of cell stiffness with every prospect of a routine application as a diagnostic tool in quantitative analysis of the physiological and pathological states of red blood cells.

[Angiosarcoma (hemangiosarcoma) cordis]. / Naczyniakomiesak krwionosny serca.

The case of a young 32 year old male with a primary cardiac angiosarcoma is reported. The neoplasm manifested itself by a quickly increasing cardiac tamponade but without metastases. The nonradical resection of the tumor was made because of local invasion with tumor tissue. The patient was followed-up for 3 years after the surgery and no recurrence of the malignant process was observed. Cardiac angiosarcoma is a very rare malignant tumor of soft tissues. In spite of significant progress in clinical treatment, for a patient diagnosed antemortem with cardiac angiosarcoma, the long-term expectations are usually very poor. Commonly known risk factors for this group of neoplasms (haemangioma of skin, chroniclymphedema, chronic post-tuberculosis pleurisy, X-ray, thorium dioxide) cannot be easily associated with the primary cardiac angiosarcoma cases. The search for chromosomal anomalies and gene mutations leading to cardiac angiosarcoma is ongoing. There is hope that recently obtained evidence for mutation of the p53gen, will provide a better understanding of this heart neoplasia.

[Epidemiological analysis of leukemia survival in Cracow for cases registered in 1980-1990]. / Ewolucja przezycia chorych na bialaczki w Krakowie--przypadków zarejestrowanych w latach 1980-1990.

The aim of the study was epidemiological analysis of survival from all types of leukemia occurring in Cracow in the years 1980-1990. The study was focused on survival times in patients according to a) cytologico-clinical type of leukemia, b) timeframe in which treatment was initiated (between 1980-1985 and 1986-1090). All patients diagnosed of leukemia between the years 1980-1990, living in Cracow and whose cytologico-clinical picture was determined had their survival times and censored survival times established. Survival until 1997 was taken into account. For each cytologico-clinical type of leukemia survival function according to Kaplan-Meier was calculated. The Cox model was implemented to analyze the risk of death depending on the period in which the disease appeared--two time frames were established 1980-1985 and 1986-1990. Other parameters considered were; age, sex and area in which the patient lived (suburb). Practically in all types of leukemia a higher probability of survival was found in patients in whom leukemia was diagnosed (and consequently treated) in the second period i.e., 1986-1990. The highest achievement was observed in acute lymphoblastic leukemia in children, in which the relative 5-year survival probability rose from 35% in the years 1980-1985 to 78% in the years 1986-1990, thus achieving the level of well developed countries. A similar picture was seen in chronic lymphocytic leukemia where the relative 5 year survival probability rose from 57% to 77%, and in chronic granulocytic leukemia where the 5 year survival probabilities were accordingly 23% and 39%. All cited values for the second period of analysis are at the levels noted in the United States and in Europe. The positive changes in the survival times observed in patients with leukemia seen in the second half of the 80-ies (in comparison to the period 1980-1985) has been interpreted as the result of advancements in therapy of the disease in Cracow.