Circulating endothelial cells, endothelial apoptosis and soluble markers of endothelial dysfunction in patients with systemic lupus erythematosus-related vasculitis.

AIM: Evidence is accumulating for endothelial cell dysfunction as one of the main factors initiating vessel wall damage in SLE. Enhanced expression of endothelial adhesion molecules is suggested to play a crucial role in the pathogenesis of vasculitis, while the number of circulating endothelial cells (CECs) is believed to be a reliable marker of endothelial damage. It therefore seems relevant to investigate CECs counts and soluble markers of endothelial dysfunction in SLE patients with inflammatory microangiopathy. The aim of this study was to assess the number of CECs, including apoptotic CECs, as well as to determine serum levels of sVCAM-1, sICAM-1 and sE-selectin in patients with SLE-related vasculitis. METHODS: The study included 51 women with SLE, divided into 2 subgroups: I patients with severe disease activity according to SLEDAI score, developing vascular complications, such as central nervous system affection and/or vasculitis and/or glomerulonephritis, II patients with mild or moderate disease activity, without vascular complications. The control group consisted of 16 healthy female volunteers. CECs, including apoptotic CECs, were isolated using anti-CD146-coated immunomagnetic Dynabeads. Serum levels of sVCAM-1, sICAM-1 and sE-selectin levels were determined with ELISA. RESULTS: In patients with SLE, CEC counts were significantly higher than in healthy controls, and strongly correlated with disease activity assessed by SLEDAI score. The number of apoptotic CECs, as compared with healthy subjects, increased considerably only in subgroup I. Serum sVCAM-1 levels were notably increased in subgroup I in relation to subgroup II and in subgroup II in relation to the control group, while serum sICAM-1 and sE-selectin levels in both subgroups were comparable and significantly higher than those of healthy subjects. CONCLUSIONS: The study showed that the number of CECs increases in SLE and strongly correlates with disease activity, reaching maximum values at the stage of inflammatory microangiopathy-related complications. Severe SLE flares are characterized by enhanced endothelial cell apoptosis. Progressive increase in serum sVCAM-1 levels is connected with disease activity aggravation and development of lupus microangiopathy.

Autonomic dysfunction in primary Raynaud's phenomenon.

AIM: The pathogenesis of Raynaud's phenomenon is still unclear. Neural and intravascular mechanisms are thought to be involved in the pathological process. The role of the autonomic nervous system is continually discussed, with particular attention to over-reactivity of the sympathetic part. The aim of this study was the clinical and electrophysiological analysis of autonomic nervous system function in patients with primary Raynaud's phenomenon. METHODS: Thirty four patients with primary Raynaud's phenomenon and 31 sex and age-matched controls were examined. Neurological examination, modified Low's Questionnaire, orthostatic and sustained handgrip tests, conduction velocity study in three nerves, sympathetic skin response (SSR), and heart rate variability (HRV) during deep breathing and at rest with the fast Fourier transform were performed. RESULTS: In the clinical examinations, 35.3% of the primary Raynaud's patients presented sensory neuropathy, but this was not confirmed in the standard conduction velocity tests. The modified Low's Questionnaire revealed dysautonomy in 82% of the patients. Autonomic regulation during the orthostatic and handgrip tests were within the normal limits. HRV at rest and the E/I ratio were significantly lower in the patient group than in the controls, while HRV spectrum analysis revealed the predominance of the low-frequency band in the patients. CONCLUSIONS: These results indicate the presence of sympathetic dysregulation and impairment of parasympathetic modulation of heart function in primary Raynaud's patients. The different cardiovascular and sudomotor functions are not affected to the same degree. These observations might support the theory of a central impairment of autonomic function in primary Raynaud's phenomenon. Peripheral nerve lesion as a coexisting cause of the observed dysautonomy remains uncertain.

Tissue factor, tissue pathway factor inhibitor and risk factors of atherosclerosis in patients with chronic limbs ischemia: preliminary study.

AIM: Thrombus formation plays a critical role in pathogenesis of cardiovascular complications in atherosclerotic peripheral arterial occlusive disease (PAOD). Tissue factor (TF) initiates the clotting cascade and is considered an important regulator of hemostasis and thrombosis. TF activity is regulated by TF pathway inhibitor (TFPI). The aim of our study was to evaluate plasma levels of the TF, TFPI and their relation to coagulation system and various other risk factors of atherosclerosis in patients with chronic limbs ischemia. METHODS: Plasma TF, total TFPI, truncated TFPI, full-length TFPI were assessed by ELISA using commercially available kits (IMUBIND Tissue Factor; Total TFPI; Truncated TFPI ELISA Kit; American Diagnostica Inc. Stamford) in 62 claudicant patients with PAOD and 20 healthy controls. RESULTS: We observed statistically higher levels of TF (94+/-52 pg/mL), total TFPI (43+/-8 ng/mL), and truncated TFPI (22+/-7 ng/mL) in patients with PAOD compared to healthy individuals (TF: 66+/-15 pg/mL; total TFPI: 36+/-4 ng/mL; truncated TFPI: 14+/-5 ng/mL). Full-length TFPI (20+/-4 ng/mL) is lower in patients with PAOD than in controls (23+/-5 ng/mL). The study indicated a positive correlation between TF and truncated TFPI (r=0.34), total TFPI and full TFPI (r=0.5), total TFPI and truncated TFPI (r=0.83) in patients with PAOD, and negative correlation between full TFPI and truncated TFPI (r=-0.65) in the control. CONCLUSION: Elevated levels of TF, disorders of balance between full-length TFPI and truncated TFPI as well as significantly increased truncated TFPI level in patients with PAOD can be independent risk factors of atherosclerotic complications.

The role of endothelin-1 and selected cytokines in the pathogenesis of Raynaud's phenomenon associated with systemic connective tissue diseases.

AIM: In the pathogenesis of Raynaud's phenomenon humoral and immunoinflammatory agents are involved. The aim of this study was the assessment of the level of endothelin-1 (ET-1), tumor necrosis factor (TNF-alpha), interleukin-6 (IL-6), soluble IL-6 receptor (IL-6sR), von Willebrand's factor (vWF) and platelet factor 4 (PF-4) in patients with Raynaud's phenomenon associated with systemic connective tissue diseases. METHODS: The examined group consisted of 32 patients (24 women and 8 men) with Raynaud's phenomenon associated with selected connective tissue diseases, aged 28-50 years. A control group consisted of 13 healthy volunteers. Immediately after a cold provocation test venous blood was taken in order to assess serum concentrations of: TNF-alpha, IL-6, IL-6sR, vWF, PF-4, antinuclear antibodies (ANA), antineutrophil antibodies (c-ANCA). RESULTS: In the group of patients with Raynaud's phenomenon mean serum concentration of ET-1, TNF-alpha, PF-4, and vWF was significantly greater than in the healthy group. In contrast, serum IL-6 and IL-6sR concentrations did not differ significantly between the diseased and healthy groups. In a subgroup of Raynaud's phenomenon patients showing particularly high concentration of serum ET-1 (twice as much as mean control concentration), the increase in IL-6, IL-6sR, vWF and c-ANCA concentration exhibited statistical significance in comparison with patients with lower serum ET-1 concentration. The vWF concentration exhibited positive correlation with time interval between the occurrence of clinical symptoms and serum ANA antibodies concentration. The increase in ET-1 synthesis in Raynaud's phenomenon patients is dependent on the increase in IL-6 level and c-ANCA antibodies level. CONCLUSIONS: The patients with Raynaud's phenomenon show an increase in ET-1 and TNF-alpha concentrations. An enhanced ET-1 synthesis is dependent on the augmentation of serum c-ANCA antibodies and IL-6 concentrations.

The biological role of protease-activated receptors in angiology. The present view.

The mechanism of initiation and growth of a thrombus in atherosclerotic arteries, although investigated extensively, has not been sufficiently elucidated. Understanding the molecular mechanisms that lead to stroke, unstable angina and myocardial infarction is of paramount importance. Protease-activated receptors (PARs) belong to a recently discovered and so far not fully characterized family of transmembrane proteins, which are involved in the initiation growth of thrombi in atherosclerotic arteries. PARs are a G-protein-coupled family of receptors which mediate a cellular function with the aid of enzymes. All of them have identical structural organization and are activated by a very similar mechanism. The enzyme (serine protease) cleaves an extracellular amino-terminal fragment of the receptor in order to unmask a new amino-terminal, 5-6 residues of which serve as the tethering ligand, able to activate its maternal receptor. According to the most recently published reports, in eukaryotic organisms there are 5 different PARs: from PAR 1 to PAR 4 found in human organisms and PAR 5 discovered as a 14-3-3 protein, identified firstly in C. elegans and Drosophila melanogaster. Up to now numerous experiments have been conducted with the aim to understand more precisely the mechanisms of PARs activation and activity. The present paper summarizes the most important and most recently published reports concerning this problem, which seems to be most relevant in angiology.

Interaction of the Pisum sativum nuclear matrix proteins with SAR DNA.

We have isolated the nuclear matrices from Pisum sativum cell nuclei using three methods: i. standard procedure involving extraction of cell nuclei with 2 M NaCl and 1% Triton X-100; ii. the same with pretreatment of cell nuclei with 0.5 mM CuSO4 (stabilisation step); and iii. method including lithium diiodosalicylate extraction. We compared the polypeptide pattern and residual DNA content of the nuclear matrices isolated. The nuclear matrices displayed a specific endonuclease activity which was due to the presence of a 32 kDa protein. The isolated nuclear matrices bound specifically the scaffold-attached (SAR) DNA derived from human beta interferon gene, in the exogenous SAR binding assay. Using the DNA-protein binding blot assay we demonstrated the presence of two nuclear matrix proteins of 66 kDa and 62 kDa which bound specifically SAR DNA.

Is an increase of deoxyribonucleases activity in uraemic lymphocytes caused by their augmented synthesis in these cells?

Deoxyribonucleases activity in T and B lymphocytes isolated from 45 patients with chronic renal failure and 30 control subjects was measured. The results obtained clearly show a dramatic increase in enzyme activity in both T and B lymphocyte cells isolated from uraemic patients when compared with the control cell. The increase in enzyme activity was limited to the group of relatively small nucleases ranging from 14 kDa to 18 kDa. Since it was shown previously that these nucleases are among the cleavage products of the largest subunit of DNA-dependent RNA polymerase class I, it is suggested that a characteristic feature of the lymphocyte cell isolated from uraemic patients is increase of RNA polymerase lability, thus resulting in RNA synthesis decrease. Amongst toxins increasing catabolism of the active polymerase enzyme, a significant correlation with the vise of lymphocyte Mg2+ and Mn2+ ions was disclosed.

Proliferative glomerulonephritis and the activity of lymphocyte DNA transcriptional enzymes.

We investigated the activity of some enzymes of the transcriptive DNA system to assess lymphocyte metabolic potential in patients with proliferative glomerulopathies. This study included analysis of the activity of three classes DNA-dependent RNA polymerases, before and 6 months after immunosuppressive therapy (Azathioprine + Prednisone). The enzyme activity was measured in nuclear extracts of T and B lymphocytes by estimation of the uptake of 3/H/UTP in the presence of alpha-amanitine. The immunoblotting technique using anti-polymerase I antibodies and nuclear proteins separation with immobilized exogenous DNA were employed to assess a type of the increase of the DNA-dependent RNA polymerase activity in lymphocytes populations (enhancement of the synthesis or limited depolymerization and degradation of the active enzyme). We found that the RNA polymerizing activity was increased in both lymphocyte populations. In T lymphocytes the increase was caused by an enhanced activity of transcript DNA enzymes, secondary to increased gene expression. In B lymphocytes an increase in enzyme activity was rather due to large stability RNA-polymerases in these cells. DNA-dependent RNA polymerase I in T and B cells was not modified by immunosuppression, while reduction in enzymatic activity of the remaining RNA-polymerase classes in T lymphocytes depends on partial limitation of their gene expression. Our study indicates that profound and various lymphocyte metabolic changes occur in patients with proliferative glomerulonephritis resulting from modifications in a gene expression and initiation of DNA synthesis in these cells.

Platelets' glycoproteins and their ligands in patients with intermittent claudication.

BACKGROUND: Platelet thrombi play critical role in pathogenesis of cardiovascular complications in atherosclerotic peripheral arterial disease (PAOD). The aim of this study was to evaluate the concentration of platelets GP IIb/IIIa, GP I b/IX and plasma levels of their ligands (fibrinogen and vWF) and their relation to other atherosclerotic risk factors in the patients with intermittent claudication secondary to PAOD. METHODS: Consecutive patients of the University Vascular Clinic were studied: 64 claudicants and 38 controls were enrolled. The concentration of platelets GPII b/IIIa and GP Ib/IX was estimated by ELISA method using monoclonal antibody against GPII b/IIIa (CD41a) and GPI b/IX (CD42a Immunotech). Plasma levels of vWF, fibrinogen, and platelets were measured by routine RESULTS: Plasma vWF (145+/-41%), fibrinogen (3.8+/-1 g/l) and platelet concentration of GP Ib/IX (121.1+/-23.39), GPIIb/IIIa (117.9 6 +/-32.7%), as well as plasma lipids and uric acid were statistically higher in claudicants than in controls (vWF: 103+/-42%, fibrinogen: 2.9+/-0.5 g/l, GP Ib/IX: 100+/-16.9%, GP IIb/IIIa: 100+/-29.4%). We have observed statistically higher concentration of GP IIb/IIIa and GP Ib/IX in smoking patients than in non-smoking patients with PAOD and significant correlation between the concentration of GP Ib/IX and GP IIb/IIIa and plasma fibrinogen in patients with PAOD and controls. CONCLUSIONS: Our results demonstrate higher platelet concentration of GP Ib/IX,GP IIb/IIIa and elevated plasma levels of ligands for platelets receptors-fibrinogen and vWF in patients with PAOD. This prothrombotic conditions may explain increased cardiovascular morbidity and mortality in this patient's group.

Coagulation in diabetic and non-diabetic claudicants.

BACKGROUND: In order to compare hemostasis in diabetic and non-diabetic claudicants we evaluated endothelial (von Willebrand factor, vWF), rheologic (fibrinogen, hematocrit), coagulation system (thrombin-antithrombin complex, TAT) and platelet (platelet factor 4, PF4, aggregation on thrombin, collagen and ADP stimulation) parameters in both groups and healthy controls. METHODS: Twenty-five diabetic, 34 non-diabetic patients with claudication and 26 healthy individuals were enrolled into the study. RESULTS: The severity of lower limbs ischemia was similar in two groups of claudicants but coronary heart disease and cerebral ischemia were significantly more common in diabetic than in non-diabetic claudicants. vWF level was significantly higher in diabetic than non-diabetic claudicants and healthy controls (184+/-43%, 147+/-43%, and 103+/-42%, respectively). Fibrinogen was significantly higher in diabetic and non-diabetic claudicants compared to controls (4.2+/-1.7, and 3.9+/-1.1, versus 2.9+/-0.5 g/l) and TAT plasma concentration was much higher in diabetic compare to non-diabetic patients and controls (9.8+/-4.4, 1.7+/-1.1, and 1.3+/-0.6 microg, respectively). PF4 concentration was significantly higher in non-diabetic patients with PAOD (34+/-29 UI/ml) when compare to healthy controls (14+/-9 UI/ml), but diabetic PAOD patients with the disease showed lower PF4 concentration (26+/-30 UI/ml). Platelet aggregation with all used activators was similar in all groups likewise hematocrit values, and platelet count. CONCLUSIONS: Complicated DM is linked with significant endothelial perturbation when compared with healthy, but also with PAOD individuals; rheologic parameters are not different from those found in PAOD patients; coagulation system activation but not platelet hyperactivity is associated with DM complicated by PAOD when compared to both control groups.

DNA-dependent RNA polymerase enzymes affected in uraemic lymphocyte cells.

The DNA-dependent RNA polymerase activity and endonucleases in uraemic lymphocyte cells were investigated. It was found that the activity and quantity in three classes of polymerases are remarkably reduced. The reduction in enzyme activity is accompanied by increasing endonuclease activity. The relationship of polymerase enzymes with endonucleases is discussed.

Deoxyribonuclease activity in T and B lymphocytes of haemodialysed patients with uraemia.

Deoxyribonuclease (DNase) activity and metal ion concentrations in lymphocytes of patients with chronic renal failure and in healthy controls were studied. The data suggest that T and B lymphocyte nuclei of patients with renal failure show increased DNase activity when compared to their healthy counterparts. It is suggested that the enhancement of enzyme activity is a result of increased metal ion concentration rather than increased enzyme copy count. The data strongly suggest that haemodialysis of uraemic patients is more effective for improvement of lymphocyte metabolism than the conservative chemical treatment.

Deoxyribonuclease activity in lymphocytes of patients with chronic renal failure treated conservatively.

The activity of nucleases and concentrations of highly important metal ions in T and B lymphocytes were examined. The source of lymphocyte was the blood of patients with chronic renal failure and activity of enzyme as well as ion concentrations were compared to the control group. Concomitant with the increase in enzyme activity was an increase of metal ion concentrations assayed in both T and B lymphocytes isolated from patients with renal disease. The data suggest that the enhancement of nuclease activity is a result of increased enzyme polypeptide synthesis and its stimulation by metal ions. Utilization of the nuclease test for monitoring uraemic toxicity is considered.

Uraemic lymphocytes: analysis of their nuclease activity.

The nuclease activity in T and B lymphocytes isolated from patients with chronic renal failure and control subjects was studied. The data obtained show a slight increase in nuclease activity in T and B cells isolated from uraemic patients as compared to the control cells. The increase in enzyme activity was limited to the group of relatively small nucleases with molecular weights ranging from 14 kDa to 18 kDa. It was documented previously that these nucleases are among the cleavage products of the largest subunit of DNA-dependent RNA polymerase I, which is responsible for ribosomal RNA synthesis. Thus, we suggest that the characteristic feature of lymphocytes isolated from uraemic patients is lowering of their metabolic activity. Amongst toxins increasing catabolism of the active enzyme, a significant correlation with the rise of lymphocyte Mg2+ and Mn2+ ions was disclosed.

[Changes in rubidium and cesium levels in the blood of patients on long-term dialysis and decreased velocity of neural conduction]. / Zmiany stezenia rubidu i cezu we krwi chorych dlugotrwale dializowanych a zwolnienie szybkosci przewodzenia nerwowego.

In 19 patients aged 19 to 45 years on long-term dialysis treatment during 12.6 +/- 18.2 months the concentrations of Rb and Cs were determined by atomic spectrometry in whole blood before and behind the dialyser at the beginning and end of dialysis. At the same time the concentrations of these elements were determined in the dialysing fluid. In all patients the velocity of conduction in the motor fibres in the upper and lower extremities was determined before and after dialysis. Damage to the peripheral neurons was demonstrated in the lower extremities mainly in 79% of cases. Increased velocity of motor conduction in at least one nerve related directly proportionally to the Cs concentration of the serum was demonstrated in 56-70% of the patients after one dialysis.

[Endothelin-1 in pathogenesis of Raynaud's syndrome]. / Endotelina-1 w patogenezie zespolu Raynauda.

Endothelin is an endogenous vasoconstrictor and plays an important role in pathogenesis of Raynaud's phenomenon. Plasma endothelin-1 (ET-1) and von Willebrand factor (vWF) concentrations following cold exposure in 52 patients with Raynaud's phenomenon were measured. Statistically significant increase of ET-1 and vWF was found in the study group in compare to healthy volunteers. There was positive correlation between ET-1 and vWF in those cases. The dates suggest that ET-changes indicates a vasospastic effect on vascular injury. Treatment with endothelin-receptor antagonist may prevent structural changes in vessel well.

[Lipoprotein and plasminogen serum levels in patients with diabetes mellitus type 2: risk factors for macroangiopathy? Preliminary study]. / Stezenie lipoproteiny (a) i plazminogenu u chorych na cukrzyce typu 2: czynniki ryzyka rozwoju makroangiopatii? Doniesienie wstepne.

Lipoprotein(a) [Lp(a)] is the specific lipoprotein which physiological role has not been explained. Similarity between apolipoprotein(a) and plasminogen structure suggests, that Lp(a) can be the bridge connecting the lipid metabolism and the coagulation system. The aim of the study was to evaluate if there is any correlation between Lp(a) and plasminogen serum concentrations in patients with diabetes t.2. 20 males and females with diabetes t.2 in the age 19-75 years (mean: 54.9 years). The control group consisted of 15 healthy men and woman in the age of 23 years (medical students). Lp(a) was estimated by ELISA, plasminogen by turbidimetric methods. Lp(a) serum level in the examined group was: 40.06 + 59.45 mg%. In 7 patients it was over 30 mg%. In the control group: Lp(a) concentration was: 13.23 + 10.5 mg%, no one was above 30 mg%. The different was significant. Plasminogen concentration was: in patients: 94.08 + 53.08%, in the control group: 89.08 + 40.7%. There was no significant difference. Correlation index between Lp(a) and plasminogen concentrations was in the patient's group: 0.3, in the control group: 0.2. In patients with diabetes we can find increased Lp(a) commentation in serum and normal plasminogen concentration. Concentration of Lp(a) didn't demonstrate significant correlation with plasminogen serum level.

[Syndrome X--open problem of contemporary medicine]. / Zespól X--otwarty problem wspólczesnej medycyny.

Based on the present literature data we have confronted the clinical parameters and some pathogenetic mechanisms of the syndrome X. Especially we stressed the danger of the accelerated development of the atherosclerotic changes in the vessels and the cardiologic complications.

[Richter's syndrome and cervical cancer in the course of a rheumatoid arthritis]. / Zespól richtera i rak szyjki macicy w przebiegu goscca przewlekle postepujacego.

The authors describe a rare case of Richter's syndrome (transition of chronic lymphatic leukaemia into reticulosarcoma) developing during rheumatoid arthritis in a women aged 53 years. Chronic antigenic stimulation and disturbances of interaction between lymphocytes T and B are suggested as a cause of Richter's syndrome.

Impact of hormonal contraception on prothrombotic activity in vessels.

One of the most common, efficient and convenient methods of contraception is hormonal contraception. Due to its popularity among young women, the safety of this method should be taken into consideration. Alternative ways of hormonal application are used and additional prothrombotic factors are being researched in order to minimize prothrombotic activity. The paper presents current data on the relationship between hormonal contraception and vascular complications based on peer-reviewed medical journals that were published between the years 2001 and 2013.