The Impact of Supplementation with Folic Acid on Homocysteine Concentration and Selected Lipoprotein Parameters in Patients with Primary Hypertension.

The aim of the study was the assessment of the impact of supplementation with folic acid on the concentration of homocysteine, total cholesterol (TC), HDL- and LDL-cholesterol, triglycerides (TG), apoprotein AI (apoAI) and apoprotein B (apoB) in patients suffering from primary hypertension. The examined group comprised 42 patients suffering from primary hypertension. All examined patients underwent laboratory tests as follows: concentration of homocysteine, folic acid, TC, LDL-cholesterol, HDL-cholesterol, TG, apoAI and apoB. All patients were orally administered with 15 mg of folic acid per day for 45 d. After this period, all laboratory tests were repeated. Homocysteine concentration was measured by the FPIA method, concentrations of apoAI and apoB were measured by the nephelometric method, and other parameters with routine methods. After administration of 15 mg of folic acid to patients with primary hypertension, a considerable decrease in the concentration of homocysteine was observed in parallel with a substantive growth of HDL-cholesterol, as well as apoprotein AI concentrations and a reduction of the apoprotein B concentration. Results of statistical analysis indicated a significant correlation between the decline in homocysteine concentration and the increase in HDL-cholesterol concentration, as well as between the increase of folic acid concentration and the increase in apoAI concentration in patients following the intake of folic acid. The drop in homocysteine concentration through the supplementation with folic acid can cause quantitative changes in the lipid and lipoprotein parameters which, in consequence, may lead to the mitigation of risk concerning the development of atherosclerosis.

Impact of administration of folic acid on selected indicators of inflammation in patients with primary arterial hypertension.

The results of epidemiological and clinical tests have shown that in patients with primary arterial hypertension, a chronic mild inflammation develops. The purpose of the study was to determine whether administration of folic acid to patients with primary arterial hypertension influences concentrations of indicators of inflammation: hsCRP, sICAM-1 and sVCAM-1. MATERIAL AND METHODS: The examination was carried out in 41 patients with primary arterial hypertension, aged 19-65 (21 men and 20 women), without complications of hypertension and/or coexisting diseases. The examined patients were administered 15 mg of folic acid once a day for 45 days. Before and after administration of folic acid, concentrations of folic acid, homocysteine, hsCRP, sICAM-1 and sVCAM-1 in serum were assessed. Concentrations of folic acid and homocysteine were determined using the immunoenzymatic method (Abbott) on an AxSYM analyzer. The level of C-reactive protein (CRP) was determined with an ultra-sensitive turbidimetric assay on a Dimension analyzer (Siemens). Next, concentrations of adhesion particles sICAM-1 and sVCAM-1 were assessed with the ELISA technique (R&D). RESULTS: After the administration of folic acid in patients with primary arterial hypertension, a significant decrease in median concentrations of homocysteine in blood was observed. Simultaneously, the median hsCRP, ICAM-1 and VCAM-1 concentrations in serum in patients with primary arterial hypertension were significantly reduced. CONSLUSIONS: Administration of folic acid to persons with primary arterial hypertension in a dose of 15 mg/ day for 45 days caused a decrease in the concentration of homocysteine in serum. That could indirectly result in the decrease in concentrations of the indicators of inflammation (hsCRP, ICAM-1 and VCAM-1), as it is apparent from previous studies that hyperhomocysteinemia stimulates the synthesis of CRP and the expression of adhesion molecules.

The clinical usefulness of assessing the concentration of cell adhesion molecules sVCAM-1 and sICAM-1 in the serum of women with primary breast cancer.

AIM OF THE STUDY: Assessment of the concentrations of the soluble forms of the cell adhesion molecules sVCAM-1 and sICAM-1 in serum of female breast cancer patients. These concentrations were assessed in relation to factors such as: age, clinical stage of disease, histological grade of malignancy, the status of the local axillary lymph nodes, and the size of the primary tumour. MATERIAL AND METHODS: A total of 103 patients with primary breast cancer, aged 29 to 89 years, were investigated. The control group consisted of 40 healthy women. The concentration of sVCAM-1 and sICAM-1 was assessed using an enzyme-linked immunosorbent assay (ELISA). RESULTS: The results of the study suggest that the level of sVCAM-1 and sICAM-1 in the serum of women with breast cancer was significantly higher than that seen in the serum of healthy women. A relationship between the level of adhesion molecules and the stage of clinical disease advancement was discovered. There was a correlation between the increasing concentrations of sVCAM-1 and sICAM-1 and with the aggressiveness of the disease. Significant differences were also found in the group of women with metastases to the axillary lymph nodes and women with no metastasis. Similar correlations were found between sVCAM-1 and sICAM-1 levels and the size of primary tumour. CONCLUSIONS: The results obtained suggest that the assessment of the soluble forms of sVCAM-1 and sICAM-1 may be useful indicators in the assessment of the clinical advancement of breast cancer.

[Hyperhomocysteinemia in patients with cardiovascular disease]. / Hiperhomocysteinemia u chorych na schorzenia ukladu krazenia

Homocysteine (Hcy) is an endogenous, non-structural protein, a sulfur-containing amino acid emerging on the pathway of methionine and cysteine, actively involved in numerous biochemical reactions. Total concentration of homocysteine in plasma of healthy humans is low and its level is between 5.0 and 15.0 mmol/l, assessed with the use of HPLC, or 5.0-12.0 mmol/l, using immunoassay methods. Higher concentration of this amino acid in blood is called hyperhomocysteinemia. Hyperhomocysteinemia is significantly correlated with cardiovascular disease and its complications: heart attacks and strokes. It is believed that hyperhomocysteinemia damages endothelial cells, reduces the flexibility of vessels, and adversely affects the process of hemostasis. In addition, hyperhomocysteinemia enhances the adverse effects of risk factors such as hypertension, smoking, and impaired glucose, lipid and lipoprotein metabolism, as well as promoting the development of inflammation. The concentration of homocysteine can be effectively lowered by supplementation with folic acid and vitamins B12 and B6. However, intervention studies conducted in the past decade did not confirm the clinical benefit of vitamin therapy lowering the level of homocysteine in blood of patients with cardiovascular disease. Moreover, there is not clear evidence from genetic studies that the presence of the gene for MTFHR polymorphism 677C>T, which is one of the most common causes of hyperhomocysteinemia, is also associated with the development of cardiovascular disease. These results led the researchers to discuss the role of homocysteine in the development and treatment of cardiovascular disease as well as the need for further research on this issue.

[Methods of preparation of the platelet-rich plasma used in medicine as an accelerator of tissue regeneration]. / Metody otrzymywania osocza bogatoplytkowego wykorzystywanego w medycynie jako preparat przyspieszajacy regeneracje tkanek.

Platelet rich plasma is being increasingly used in the modem medicine as a material, stimulating regeneration and accelerating tissue healing. Platelet rich plasma is an autologous platelet concentrate, which is obtained from the peripheral blood of the patient. The method of extraction is based on the isolation of platelets during centrifugation of the whole blood, drew on anticoagulant. With the difference in density between the various cellular components of blood, such as red blood cells, buffy coat and platelet poor plasma, the separation into individual fractions is possible. At the present moment no optimal method of preparation of the platelet rich plasma has been found. On the market there are a number of commercial collection systems available, differing from each other in centrifugation parameters, type of container to which blood is collected and anticogulant used. Unfortunately, this can lead to obtaining platelet rich plasma with a varying number of platelets, leukocytes and resulting in a different concentration of growth factors. This is important, because the studies show, that a positive clinical effect depends on the quality of the used platelet-rich plasma.

Hyperhomocysteinemia, lipid and lipoprotein disturbances in patients with primary hypertension.

PURPOSES: The main aim of the study was to answer two questions: what are the concentrations of total cholesterol, LDL-cholesterol, HDL-cholesterol, triglycerides, apo A-I (apolipoprotein A-I), apo B (apolipoprotein B) and Lp(a) (lipoprotein(a)) in serum of patients with primary hypertension and with hyperhomocysteinemia? Is there any correlation between the concentration of homocysteine in blood serum and investigated lipid and lipoprotein parameters in patients with primary hypertension? MATERIAL/METHODS: We investigated 42 patients with primary hypertension, aged 22-57. The control group consisted of 20 healthy volunteers. The concentration of homocysteine in serum was evaluated using immunochemical method (FPIA - Fluorescence Polarization Immunoassay). The concentration of total cholesterol, LDL-cholesterol, HDL-cholesterol and triglycerides in blood serum were estimated using enzymatic method. Apo A-I, apo B and lipoprotein(a) were assessed using nephelometric method. RESULTS: The analysis of the results revealed statistically significant lower concentrations of HDL-cholesterol and apo A-I in blood serum of patients with primary hypertension and hyperhomocysteinemia than in the population with hypertension and normohomocysteinemia. Negative correlation between homocysteine and HDL-cholesterol as well as apo A-I has been revealed. CONCLUSION: Quantitative analysis of the concentration of lipids and lipoproteins in blood serum in patients with primary hypertension and hyperhomocysteinemia may suggest that this type of human population might be more susceptible to atherosclerosis than those with primary hypertension and normal values of homocysteine.

[Endothelial dysfunction in patients with primary hypertension and hyperhomocysteinemia]. / Dysfunkcja sródblonka naczyniowego u chorych na pierwotne nadcisnienie tetnicze z hiperhomocysteinemia.

It is widely accepted that endothelial dysfunction is the basis of the development of cardiovascular diseases, including hypertension. With regard to hypertension, endothelial dysfunction is concerned mainly with impaired vascular expansion; however, it is also related to the intensity of the development of atherosclerosis and thrombosis. Among the factors that cause damage to the endothelium, along with classic risk factors, is hyperhomocysteinemia. Hyperhomocysteinemia promotes the formation of oxygen radicals, lowering the oxidation-reduction potential, adversely affects the biosynthesis and function of vasodilator factors in the vascular wall, contributes to the inhibition of endothelial cell division with intense myocyte proliferation and migration, and impairs production of extracellular matrix components in the vascular wall. In addition, high levels of homocysteine and its derivatives contribute to the modification of LDL and HDL particles, inflammation and disorders in coagulation and fibrinolysis. Biochemical effects of the impact of hyperhomocysteinemia on endothelium can lead to damage of endothelial cells, dysfunction of diastolic function of vessels and reduction of their flexibility through its influence on vascular wall remodeling. These changes lead to an increase in blood pressure, strengthening the development of hypertension and target organ damage in patients with this disease.

Clinical usefulness of assessing VEGF and soluble receptors sVEGFR-1 and sVEGFR-2 in women with breast cancer.

INTRODUCTION: The biological activity of VEGF depends on the presence of its specific receptors on the endothelial surface: VEGFR-1, VEGFR-2, and on their soluble forms sVEGFR-1 and sVEGFR-2. The binding of the membrane-bound receptors with VEGF affects the permeability, proliferation and migration of vascular endothelial cells. This creates the necessary conditions for the vascularisation of solid tumours and for the spread of remote metastases. The sVEGFR-1 and sVEGFR-2 receptors are believed to be natural inhibitors of VEGF. OBJECTIVE: To determine the clinical usefulness of VEGF and the sVEGFR-1 and sVEGFR-2 receptors level assay in women with primary breast cancer. The assessment also took into account: patient's age, stage of the disease, histological grade, status of the axillary lymph nodes and size of the primary tumour. MATERIAL AND METHODS: The concentrations of VEGF, sVEGFR-1 and sVEGFR-2 were ascertained in 103 women with primary breast cancer. The concentrations of VEGF in the plasma, and those of the soluble receptors sVEGFR-1 and sVEGFR-2 in the serum, were assessed by ELISA, R&D Systems. RESULTS: The study found significantly raised concentrations of VEGF, sVEGFR-1 and sVEGFR-2 in the serum of women with breast cancer, relative to the values obtained from the control group. It was found that with increasing clinical stages of the disease, the levels of VEGF and concentrations of sVEGFR-1 and sVEGFR-2 also increased. Similar findings were noted when assessing the degree of the histological grade of the tumours. Significantly higher values of VEGF protein and the assessed receptors were obtained from women with metastases to the axillary lymph nodes. A positive relationship, though without statistical significance, was noted between the concentration of sVEGFR-2 and the size of the tumour. CONCLUSIONS: The high concentrations of the VEGF cytokine and the sVEGFR-1 and sVEGFR-2 receptors in women with breast cancer are responsible for giving rise to the processes of tumour angiogenesis. The concentrations of the VEGF protein and the soluble forms of the receptors sVEGFR-1 and sVEGFR-2 in the serum of breast cancer patients showed positive correlations with the clinical stage of the disease. These results point to the usefulness of VEGF assessment and its soluble receptors in the clinical evaluation of patients with breast cancer.

High concentration of urokinase-type plasminogen activator receptor in the serum of women with primary breast cancer.

AIM OF THE STUDY: The purpose of this study was to assess the concentration of urokinase-type plasminogen activator receptor (uPAR) in the serum of 103 women with breast cancer. Commonly recognized prognostic factors were taken into account, including age, histological grade of malignancy, stage of clinical advancement of the disease, status of local axillary lymph nodes and the size of the primary tumour. MATERIAL AND METHODS: The concentration of uPAR was assessed using an enzyme-linked immunosorbent assay (R&D Systems). RESULTS: The concentration of uPAR in women with breast cancer was found to be higher than in a control group and the difference was statistically significant. The concentration of uPAR was found to increase in line with increasing disease stage and this too was of statistical significance. Raised levels of uPAR were found in women with breast cancer both with and without metastases to the lymph nodes of the axilla. A positive relationship was also found between the concentration of the tested receptor and the size of the primary tumour. No significant relationship, however, was found between the concentration of uPAR and the histological grade of malignancy of the tumour. No statistically significant results were obtained regarding the menopausal status of the women, that is, whether they were pre- or post-menopausal. CONCLUSIONS: Concentration of uPAR in serum of women with breast cancer is positively correlated with the stage of advancement of the disease. Thus, the assessment of this parameter can be useful in the clinical evaluation of women with breast cancer.

The determination of VEGF and MVD, among patients with primary breast cancer.

The purpose of the study was to ascertain the value of assessment of vascular endothelial growth factor (VEGF) levels and microvessel density, and to search for correlations between them, in women with breast cancer. The assessment considered factors such as the stage of clinical disease advancement--according to International Union Against Cancer, the grade of histological malignancy, status of axillary lymph nodes and the size of the primary tumour. The concentration of VEGF was assessed in the plasma of 103 women with breast cancer, using an immunoenzymatic method (Quantikine test of R&D Systems). Assessment of microvessel density was performed using histopathological immunoperoxidase methods, using an anti-von Willebrand factor antibody (DAKO A/S). A statistically significant relationship was found between rising VEGF levels and microvessel density in women with breast cancer, when compared to values from a control group. A correlation was found between VEGF concentration and microvessel density (MVD) values. Statistically significant differences were found between VEGF levels of patients in stages I, II and III of clinical disease advancement. For MVD, differences were found only between stages I and III. A statistical relationship was also found between VEGF and MVD and tumour size. Similar results were found between VEGF concentrations in women with metastases to the axillary lymph nodes and cytokine levels in women with no metastases. The results of the study suggest that the degree of tumour vascularization and the concentration of VEGF may represent valuable indicators for the assessment of the angiogenic process in women with breast cancer.

[Zymography--method for quantitation of activity on gelatinase A (pro-MMP-2, 72 kDa) and gelatinase B (pro-MMP-9, 92 kDa) in serum of patients with breast cancer]. / Zymografia--metoda oceny aktywnosci zelatynazy A (pro-MMP-2, 72 kDa) i zelatynazy B (pro-MMP-9, 92 kDa) w surowicy chorych na raka sutka.

UNLABELLED: Zymography is an electrophoretic method for measuring proteolytic activity. The method is based on polyacrylamide gels impregnated with a protein substrate (gelatin, casein), which is degraded by the proteases. It is already widely used for research on extracellular matrix degrading enzymes, in particular the matrix metalloproteinases (MMPs). The aim of this study was to evaluate the zymography used to estimate gelatinase A (pro-MMP-2, 72 kDa) and gelatinase B (pro-MMP-9, 92 kDa) in serum of women with breast cancer. MATERIAL AND METHODS: The study was conducted on the serum of 90 female with breast cancer aged 32-85 years (average 57.2 year) and in a group of 30 women with benign breast diseases aged 34-87 years (average 55.4 year). RESULTS: The results showed significantly higher activity ofgelatinase A and gelatinase B in the group of women with breast cancer that in the control group. The activity of gelatinase A was over 0.48 AU/ml in 50 women (55.6%) and gelatinase B in 46 women in this group (51.1%). Further analysis showed a strong correlation between activity of gelatinase A and B and advance stage of breast cancer, lymph node status, and tumour burden. An elevated activity of gelatinases in patients with breast cancer confirms the role of these enzymes in the development of such tumours. There was no significant difference in the gelatinases activity between the serum samples from patients with breast cancer and those from women with benign breast diseases. The diagnostic sensitivity of this procedure used to estimate gelatinase A and gelatinase B activity carried out 56% and 51%, and the diagnostic specificity 75% and 77%, respectively. CONCLUSIONS: Zymography is a simple, sensitive, and functional assay for analysing proteolytic activity of gelatinase A and gelatinase B. The diagnostic accuracy of gelatinases evaluation is limited by the lack of sensitivity and specificity in early stage of the disease and in differentiation of breast cancer from benign diseases. Measuring their activity in serum breast cancer patients indicates the patient's clinical condition. A good correlation between serum activity of gelatinases and the prognostic factors suggest its usefulness as a marker both in the follow-up and in the prognosis of breast cancer patients.

Diagnostic value of measuring serum CA 15-3, TPA, and TPS in women with breast cancer.

INVESTIGATION: The concentrations of CA 15-3, TPA, and TPS were determined in the serum of 90 women with breast cancer MATERIAL/METHODS: Serum CA 15-3 levels were determined by Abbott immunofluorescent tests, TPA concentration using the Byk-Roland immunoluminometric method, and TPS by the BEKI Diagnostics immunoenzymatic method. RESULTS: In the diagnosis of breast cancer, the TPA test had the greatest diagnostic sensitivity. A strong correlation was found between the percentages of women with positive levels of all three markers and advanced stages of breast cancer. A longitudinal evaluation of 42 women who underwent neoadjuvant chemotherapy showed clinical regression of the tumor and a statistically significant reduction in the concentrations of these markers CONCLUSIONS: These results indicate that determining the serum levels of CA 15-3, TPA, and TPS may be used to establish the stage of disease and it provides useful guidance both in deciding whether to treat surgically and in estimating the efficacy of neoadjuvant chemotherapy in patients with breast cancer.

Alterations in skeletal protein, distribution of PKCalpha, and level of phospholipids in erythrocyte membranes of women with primary breast cancer.

The aim of our work was to study the influence of primary breast cancer on mature erythrocyte membranes. Blood was sampled from 29 women with primary breast cancer, aged 35-86 years, in different stages of clinical progression of the disease. In red blood cell membranes an increase of phosphatidylinositol-4-phosphate and phosphatidylinositol-4,5-diphosphate levels was observed. These changes were accompanied by a decrease in phospholipase C activity. Simultaneously, a significant decrease in concentration of phosphatidylserine, sphingomyelin, and phosphatidylinositol was found. Quantitative protein evaluation showed an increase in band 4.1 protein content with no changes in the level of constitutive PKCalpha responsible for the phosphorylation of this protein and its affinity to glycophorine C. In parallel a greater increase of PKCalpha translocation after PMA treatment compared to controls was observed. Possible oxidative damage of erythrocyte membranes indicated by an increase in malonyldialdehyde level and decrease in SH-group content as well as by an increase in the w/ ratio was documented. From the results it is concluded that primary breast cancer seems to affect the membranes of mature erythrocytes.

[Concentration of estrogen and progesterone receptors in normal and neoplastic tissue in patients with laryngeal neoplasm]. / Stezenie receptorów estrogenowo-progesteroneowych w tkance zdrowej i nowotworowo zmienionej u chorych na raka krtani.

The analysis of larynx cancer localisation inside the larynx and hypopharynx indicate that there are some places prone to cancerogenesis. Differences in morbidity, organ localisation and prognosis of larynx cancer can be explained not only by exogenic factors but also by endogenic ones, including hormones. The estrogen (ER) hormone receptor concentration and progesterone (PgR) concentration was estimated in the neoplastic tissue in 18 patients (16 men, 2 women). In the control group, in 10 patients (9 men, 1 women) the concentration receptor was assessed in surrounding, macroscopically unchanged tissue. The quality assessment was done both in cytoplasmatic and nuclear receptors. 6 specimens of neoplastic tissue was taken from vocal folds, 7 from vestibular folds, 2 from subglottic space, 2 from epiglottic petiolus and 1 from pyriform sinus. Control group consist of 10 healthy tissue specimens including 4 taken from the vocal cords. 4 from vestibular folds, 2 from free edge of epiglottis. All samples were examined by pathologist. The concentration of cytoplasmatic and nuclei receptors (ER and PgR) was estimated by immunoenzymatic essay by ER-EIA and PgR-EIA monoclonal antibodies (Abbott). The highest concentration of receptors in larynx cancer was stated in tissue coming from vestibular folds and epiglottic petiolus in healthy objects in the region of vestibular folds.

Significance of the evaluation of the tissue proliferation specific marker (TPS) and the carcinoembryonal antigen (CEA) in patients with ulcerative colitis and adenocarcinoma.

BACKGROUND: The aim of our research was to evaluate TPS concentration as a marker of proliferative processes and the levels of CEA in children and adults with ulcerative colitis (UC) and in adults with colorectal adenocarcinoma (AC), to find out if there is any interdependence between TPS and CEA concentrations in these patients and to evaluate the usefulness of TPS as a marker of neoplastic risk in patients with UC. MATERIAL/METHODS: The study was carried out in 3 groups of patients: Group I consisted of 15 children, ages 10-18, treated for UC; Group II, 22 adults, ages 23-40, treated for UC; and Group III, 14 patients, ages 40-60, in whom AC was diagnosed. RESULTS: In Group II, the mean TPS concentration was significantly higher than in Group I (p<0.00001). In Group III, adults with AC, the mean TPS concentration was 1074.00+/-1356.87 U/l. Thus there was a statistically significant difference between TPS concentrations in adults with UC and adults with AC (p<0.00005). There was no statistically significant difference between CEA levels in children and adult groups with UC. However, a statistically significant difference occurred between CEA concentrations in children and adults with UC and the group of adults with AC (p<0.00001). CONCLUSIONS: It would appear that testing for TPS concentration in patients with UC is helpful in monitoring the proliferative process, whereas in patients with neoplastic processes it serves as a screening examination, enabling early detection of recurrences, full evaluation of neoplastic growth, and evaluation of therapeutic effectiveness.

[Correlation of sex hormones in blood serum and estrogen-progesterone receptors concentration in patients with laryngeal cancer]. / Korelacja stezenia hormoneów plciowych w surowicy krwi ze stezeniem receptorów estrogenowo-progesteronowych u chorych na raka krtani.

Particular feature of larynx cancer epidemiology is large difference in morbidity at man and woman. It can be explained not only by exogenic factors, but also by endogenic once, including hormones. There was estimated estradiol and progesterone value in blood serum. The examined group comprised 12 larynx cancer patients. It was used radioimmunological assay Spectria by Orion Diagnostica. At the same time the estrogen and progesterone hormone receptor concentration was estimated in the neoplastic tissue in 18 patients. In the control group, in 10 patients the concentration was examined in surrounding, macroscopically unchanged tissue. The concentration of receptors was estimated by immunoenzymatic assay ER-EIA and PgR-EIA by Abbott. The quality assessment was done both in cytoplasmatic and nuclear receptors. There was performed correlation between hormone value in blood and receptor concentration in larynx cancer tissue at 12 patients and in non-neoplastic tissue at 5 patients. There were many abnormal steroid hormones value in blood serum. We found positive correlation between estradiol and progesterone value in blood and concentration of their receptors in larynx cancer. There were observed anomalies of hormone-receptor system at larynx cancer patients.