Circulating endothelial progenitor cells are inversely correlated with in-stent restenosis in patients with non-ST-segment elevation acute coronary syndromes treated with EPC-capture stents (JACK-EPC trial).

AIM: Aim of the study was to evaluate the association between circulating endothelial progenitor cells (EPCs) and angiographic outcomes after implantation of GenousTM stent in patients with non-ST-segment elevation acute coronary syndromes (ACS) (NSTE-ACS) undergoing urgent percutaneous coronary intervention (PCI). METHODS: Sixty patients treated with EPC-capture stent (N.=30) or bare metal stents (BMS) (N.=30) receiving 80 mg atorvastatin and dual antiplatelet therapy (DAT) for 12 months. Restenosis was assessed after 6 months by quantitative coronary angiography (QCA) and major acute coronary events (MACE) evaluated after 6 and 12 months. INCLUSION CRITERIA: de novo lesion >70% in native vessel, diameter 2.5-4 mm, lesion length <30 mm. EXCLUSION CRITERIA: diabetes, previous revascularization, significant left main stenosis, chronic total occlusions (CTO) and multivessel disease. RESULTS: Majority of patients in EPC-capture stent and BMS groups presented with NSTEMI (73.3% and 70%, respectively). Mean stent length was 20.1±8 and 19.9±10 mm, diameter 3±0.97 and 3.1±0.88 mm in respective groups. The binary restenosis was significantly lower in GenousTM (13 vs. 26.6%, P=0.04). Risk of MACE after 6 and 12 months were comparable in both groups. There was no stent thrombosis. Numbers of circulating EPCs were significantly approximately 2-fold higher during the ACS than after 6 months. Mobilization of EPCs during acute ischemia was significantly lower in patients who developed restenosis after 6 months (3 vs. 4.5 cells/µL, P=0.002) and it was negatively correlated with late-loss after 6 months (R=-0.42; P<0.03). CONCLUSION: Use of GenousTM stents in NSTE-ACS is associated with lower restenosis rate than BMS at 6 months. There was no ST through 1 year. The number of circulating EPCs is inversely correlated with in-stent late loss (LL).

Changes in circulating microRNA levels associated with prostate cancer.

BACKGROUND: The aim of this study was to investigate the hypothesis that changes in circulating microRNAs (miRs) represent potentially useful biomarkers for the diagnosis, staging and prediction of outcome in prostate cancer. METHODS: Real-time polymerase chain reaction analysis of 742 miRs was performed using plasma-derived circulating microvesicles of 78 prostate cancer patients and 28 normal control individuals to identify differentially quantified miRs. RESULTS: A total of 12 miRs were differentially quantified in prostate cancer patients compared with controls, including 9 in patients without metastases. In all, 11 miRs were present in significantly greater amounts in prostate cancer patients with metastases compared with those without metastases. The association of miR-141 and miR-375 with metastatic prostate cancer was confirmed using serum-derived exosomes and microvesicles in a separate cohort of patients with recurrent or non-recurrent disease following radical prostatectomy. An analysis of five selected miRs in urine samples found that miR-107 and miR-574-3p were quantified at significantly higher concentrations in the urine of men with prostate cancer compared with controls. CONCLUSION: These observations suggest that changes in miR concentration in prostate cancer patients may be identified by analysing various body fluids. Moreover, circulating miRs may be used to diagnose and stage prostate cancer.

Liposome encapsulated aurothiomalate reduces collagen-induced arthritis in DBA/1J mice.

Collagen-induced arthritis (CIA) generated in rats or mice has long been a model system for the study of rheumatoid arthritis in humans. In particular, this system has been used to study the mechanisms and effects of anti-arthritic drugs in the treatment of the disease. Sodium aurothiomalate (ATM) is an agent often used to treat rheumatoid arthritis in humans; however, it possesses inherent toxicities which limits its usefulness. Liposome-encapsulated drugs are currently being developed to minimize the toxicities associated with a variety of potentially beneficial drugs. We have chosen to encapsulate ATM into small unilamellar vesicles (SUVs) to determine whether greater efficacy would be achieved in treating CIA with SUV ATM as compared to using the free drug. SUVs were prepared from hydrogenated egg phosphatidylcholine and cholesterol. These SUVs were very stable. Vesicles stored at 4 degrees C lost only 0.09% of encapsulated ATM (SUV ATM) after 14 days and were able to reduce collagen-induced arthritis in these mice. Animals treated by i.m. injections of SUV ATM exhibited a 50% reduction in symptoms. More importantly, histological examination of knee joints of the affected animals verified that SUV ATM treatment prevented cellular infiltration of lymphocytes into the synovia of the collagen-sensitized mice. Conditioned media from spleen cell cultures was assayed for the presence of inflammatory lymphokines that might be affected by SUV ATM to account for the success in suppressing collagen-induced arthritis.

Interleukin-6-type cytokines and their receptors for gene therapy of melanoma.

B-78-H1 melanoma cells were stably transfected with cDNAs encoding human IL-6, human LIF, murine sIL-6R and murine sLIFR. The mock transfected and transfected cells demonstrated no detectable H-2Kb molecules. B-78 transfected cells were subcutaneously (s.c.) and intravenously (i.v.) injected to B57BL/6 x C3H mice. Control B-78 cells formed tumors and lung metastases in injected animals. Cells transfected with IL-6, LIF and sIL-6R showed greatly reduced tumor and metastases formation. Transfection of IL-6, sIL-6R or LIF had similar protective effects while the combination of IL-6 and sIL-6R was most effective. In contrast, cells transfected with sLIFR showed reduced metastasis formation but increased tumor growth compared to mock transfected cells. Kinetic analysis demonstrated a 3 weeks lag period between the formation of tumors by B-78 cells and the combination of B-78 cells transfected with IL-6 and sIL-6R. No such lag phase was seen when B-78-IL-6 or B-78-sIL-6R cells were injected alone. Mice primarily injected s.c. with a mixture of IL-6 and sIL-6R transfected cells and rechallenged after 2 weeks with parental B-78 cells demonstrated long-lasting antitumor immunity. IL-6 and sIL-6 transfected cells used alone for immunization had only limited effect. Injection of transfected cells into SCID mice which are characterized by greatly reduced number of T and B cells, showed a protective effect of sIL-6R on metastasis formation by B-78 cells. beta 2m knockout mice lacking CD8+ T cells, injected with B-78 cells developed tumors and died after 2 weeks. However, B-78 cells transfected with IL-6 developed tumors in only 50% of animals. Mice without tumors rechallenged with B-78 cells demonstrated required immunity against parental melanoma cells. The results obtained indicate that studied IL-6-type cytokines and their respective soluble receptors affect murine melanoma growth and metastasis formation. The major finding of these studies is that IL-6 complexed with sIL-6R demonstrated qualitatively different biological activity than IL-6 alone especially in stimulating long lasting anti-melanoma immunity. The proposed mechanism of action of such complexes beside activation of cytotoxic T lymphocytes is activation of NK cells.

Some aspects of the copper(II)-DNA interaction.

The Cu(II) ion interaction with calf-thymus DNA was studied by means of differential pulse polarography and sweep voltammetry as well as chromatography and viscosimetry. Most of the complexes formed at high ionic strength (0.2 M) and lower Cu(II) concentrations are of a nondenaturing nature. Their formation has but a minor effect on unwinding process of the DNA double helix. The excess of Cu(II) (P = 5) leads, however, to distinct denaturation of the DNA structure. Metal ions have little effect on the denaturation induced by the polarographic reduction of DNA on the mercury electrode. This conclusion is consistent with the character of the polarographic process and with the fact that Cu(II) ions are not very effective in the interaction with AT pairs. Cupric ions have no renaturing ability towards thermally denatured DNA at 0.2 M ionic strength but distinct renaturation was observed at low ionic strength (0.05 M).

Microheterogeneity forms of alpha 1-acid glycoprotein as indicators of rheumatoid arthritis activity.

The microheterogeneity of alpha 1-acid glycoprotein (AGP) has been studied in the sera of 48 patients with rheumatoid arthritis and of 12 healthy individuals. For each rheumatoid patient the disease activity has been assessed and each patient has been assigned to one of four activity grades: I, inactive; II, mildly active; III, moderately active; and IV, severe. Concanavalin A-affinity electrophoresis revealed three microheterogeneity variants of AGP: non-reactive with Con A, weakly reactive with Con A and strongly reactive with Con A. The relative amounts of AGP-variants observed in the healthy donors were similar to those observed in the patients with activity grade I, but differed significantly from patients with grades II, III and IV. The differences between the AGP-patterns of patients with activity grades II, III and IV were also statistically significant.

Binding of C-reactive protein (CRP) by human peripheral blood lymphocytes in vivo.

Antigenicity of C-reactive protein (CRP) on the surface of human lymphocytes was investigated by use of indirect immunofluorescence technique with anti-CRP antibodies. CRP on the lymphocyte surface (sd-CRP) belongs to two different categories: i) CRP produced by lymphocytes and inserted into cell membrane (s-CRP), ii) CRP produced primarily by the liver and bound by the lymphocytes (sb-CRP) in calcium-dependent manner. In human peripheral blood of healthy donors approximately 2.5% of lymphocytes expressed membrane CRP (s-CRP) and 1.5% of lymphocytes bound CRP in calcium-dependent manner (sb-CRP). Percentage of s-CRP lymphocytes increased in patients with rheumatoid arthritis, while population of sb-CRP lymphocytes did not change significantly, except cases where serum CRP concentration reached more than 50 micrograms/ml. Thus, it can be concluded that CRP is bound to the distinct population of lymphocytes, bearing specific membrane receptors.

The modified method for isolation and culture of highly homogeneous Leydig cell population from rat testes.

A modified method for isolation and culture of a pure population of rat Leydig cells is described. For obtaining crude interstitial cell suspension, decapsulated testes were dispersed in 0.02% collagenase solution in Ca2+, Mg2+--free Hanks medium for 1 hour. Then, approx. 5 X 10(7) cells were centrifuged in 10-90% discontinuous, isoosmotic Percoll gradient at 3000 g for 20 min. The cells from eight fractions obtained were collected and cultured in Eagle's MEM for 4 days. Using morphological methods, 1.059-1.070 g/ml density fraction contained 97% and 1.070-1.080 g/ml fraction contained 90% viable Leydig cells. The cells secreted testosterone to the culture medium and responded to LH stimulation with over four-fold increase in hormone secretion.

The influence of subcutaneously administered lead(II) acetate on the concentrations of copper, iron, and zinc in the blood, kidney, liver, and spleen of rats.

The concentrations of copper, iron, lead, and zinc in the blood, kidney, liver and spleen were determined before and after subcutaneous administration of lead(II) acetate (100 mg Pb kg-1 body weight) to male albino Wistar rats. The control rats had the following concentrations (microgram g-1 dry weight) of Cu, Fe, Pb and Zn: blood, 4.5, 3200, 2.1, 182; kidney, 36, 585, 120, 92; liver, 11.1, 720, 14.3, 124; spleen, 4.5, 420, 7.0, 76. After administration of lead, rats were sacrificed after 12, 24, 48, 72 and 96 h. The Pb concentration in the blood remained constant for the first 24 h at the level of the control group (2.1 micrograms g-1) and had decreased to half that level at 96 h. The lead concentrations peaked in the organs at 110-142% of those in the control group and had decreased at 96 h to levels considerably below those in the control group. The concentrations of Cu, Fe and Zn increased in the three organs to values 117-161% found for the control group. At 96 h the concentrations of Cu, Fe and Zn in the spleen had returned to levels of the control group; the concentrations in the liver were 112-153%, and in the blood 89-93% of those of the control group. In the kidney the iron (110%) and the zinc (126%) concentrations at 96 h were higher than the control values, whereas the copper concentration was the same as the control value. The concentrations in the blood were least affected. The most drastic changes were observed in the liver.(ABSTRACT TRUNCATED AT 250 WORDS)

Pathways to psychiatric care and reform of the public health care system in Poland.

OBJECTIVE: - Since 01.01.1999 a reform of the health care system in Poland has been realised based on the general health insurance act. We attempt to use the pathway method, especially measuring of time intervals along pathways, as a way of making accessibility to mental health services operational. METHOD: - In a period of 2 months all patients aged 15 and over who had not sought care from any public or private psychiatric service during the previous 365 days, were interviewed by psychiatrists, using the Polish version of the WHO Encounter Form. RESULTS: - A total of 228 patients were seen. The study reveals that the median interval between first seeing a primary care giver and arrival at a mental health service was 12 weeks, which is much longer than all other European centres in previous studies. CONCLUSION: - Making use of the pathway method and, in particular, median interval analysis between the onset of the mental health problem and seeing mental health professionals (MHPs) has shown practically limited access of patients, with a new episode of care to psychiatric services in both epidemiological catchment areas. This method seems to be a simple and inexpensive way of monitoring the accessibility to MHP in the period of health care reform.

[Alpha-1-acid glycoprotein in differential diagnosis of rheumatic diseases]. / Das alpha-1-saure Glykoprotein in der Differentialdiagnose rheumatischer Erkrankungen.

In recent years, the investigation of acute-phase proteins with the aid of affinity electrophoresis employing lectins as carrier substance, has become ever more important, in particular in the diagnosis of inflammatory rheumatic diseases. In order to investigate the usefulness of a determination of alpha-1 glycoprotein microheterogeneity in the sera as a diagnostic parameter, we evaluated, on a prospective basis, the sera of 85 consecutive patients presenting at our department with various inflammatory rheumatic diseases (chronic rheumatoid arthritis [RA] [n = 22], seronegative spondarthropathies [SpA] [n = 15], polymyalgia rheumatica [PMR] [n = 10], polymyositis dermatomyositis [PM/DM] [n = 8], osteoarthritis [n = 18], and infectious diseases [n = 12]). The results were expressed as reactivity coefficient of the alpha-1 acid glycoproteins (AGP/RC). A significant increase in the AGP/RC was observed in patients with various infections, and in those with RA or SpA with intercurrent infection, as compared with patients with RA or SpA with no intercurrent infection, or in healthy controls. It also proved possible to differentiate between PM/DM and PMR. Determination of the erythrocyte sedimentation rate, and quantification of the C-reactive protein and of alpha-1 glycoprotein, permitted no discrimination between an increase in inflammatory activity related to the basic disease, and an intercurrent bacterial infection. The results show that the determination of qualitative changes in the acute phase proteins, in particular alpha-1 acid glycoproteins, may make it possible to differentiate between inflammatory and infectious diseases. This examination technique may be of future clinical importance.

[Angio-ultrasonography--a method of examining the extracranial arteries]. / Angioultrasonografia--metoda badania tetnic pozaczaszkowych.

The authors describe a method of investigations of the extracranial cerebral arteries--angioultrasonography based on the Doppler effect. The study was done with a Polish apparatus designed and produced by the Experimental Laboratory of the Institute of Basic Technical Problems, Polish Academy of Sciences. The study confirmed the usefulness of this method in the diagnosis of arterial diseases involving the extracranial arteries supplying the brain. A high agreement was demonstrated between the results of angioultrasonography and classical angiography. Owing to the use of a grey scale it was possible to localize accurately the site of stenosis or occlusion of the studied artery.

[The evaluation of the coronary artery sufficiency in myocardial infarction patients after cardiac surgery and preventive therapy]. / Ocena wydolnosci wiencowej u chorych po zawale serca leczonych operacyjnie lub zachowawczo.

During 5 years period (1989-1993) the authors investigated a group of 40 patients with coronary artery disease after myocardial infarction treated in the Silesian Center of Cardiology in Katowice. 20 patients were operated on (CABG), 20 were medically-treated. It was evaluated the history, physical status, stress-test, echocardiography and 24-hours ECG. Stress test was estimated according to Mark's test. In the echocardiographic examination it was observed wall motion score index (WMSI) and the left ventricular abnormal contraction area (AA). In the operated group it was noticed higher physical ability and no influence of CABG on left ventricular contractability.

[Vagal nerve neurinoma: a case report]. / Nerwiak nerwu blednego--opis przypadku.

A case of asymptomatic 44-year-old male patient with moderate arterial hypertension, misdiagnosed as aortic aneurysm, is reported. Additional diagnostics (transthoracic and transesophageal echocardiography, CT scan) revealed the presence of mediastinal tumor, diagnosed at operation as vagal neurinoma. Early and late results of therapy are described.

Circulating microRNAs (miR-423-5p, miR-208a and miR-1) in acute myocardial infarction and stable coronary heart disease.

AIM: The microRNAs (miRs) are small non-coding RNAs which regulate expression of multiple genes involved in atherogenesis. MicroRNA are also present in circulation. The aims of this study were: 1) assessment of expression level of miR-1, miR-208a and miR-423-5p in plasma in patients with STEMI, stable CAD and healthy individuals; 2) evaluation of correlation between plasma miRs and left ventricle ejection fraction, end- systolic and end-diastolic diameters and troponin release in patients with STEMI. METHODS: Study group consisted of 26 patients: 1) acute MI group (N.=17); 2) stable CAD group (N.=4); and 3) subjects with no history of CAD (control group, N.=5). Expression of miR-423-5p, miR-208 and miR-1 was measured in plasma before PCI, 6, 12 and 24 hours later. Expression level ofmiRs was measured using TaqMan® MicroRNA Assays. Expression was assessed by Pfaffl method, and miR-39 was used for normalization of the results. RESULTS: In stable CAD in comparison to control group the expression level of miR-1, miR-208a and miR-423-5p did not show significant differences. Also there was no significant increase of number of miR copies at 6, 12 and 24 hours after PCI. There was a significantly higher number of miR-423-5p copies in patients with acute MI before the pPCI. After 6, 12 and 24 hours post-procedure the expression level was similar to the control group and significantly lower than the baseline level. Conversely, the expression level of miR-1 and miR-208a were not significantly different than in the control group. In patients with acute MI there were no significant correlations between the expression level of miRs and any of the echocardiographic parameters of LV as well as level of troponin I at any time-point of the follow-up. CONCLUSION: Early in acute myocardial infarction the expression of miR-423-5p in plasma is significantly increased with subsequent normalization within 6 hours. Potentially it is an early marker of myocardial necrosis.

Use of the Hypomania Checklist-32 and the Mood Disorder Questionnaire for detecting bipolarity in 1051 patients with major depressive disorder.

PURPOSE: To use the hypomania checklist (HCL-32) and the mood disorder questionnaire (MDQ), for detecting bipolarity in depressed patients. PATIENTS: One thousand and fifty-one patients fulfilling ICD-10 criteria for unipolar major depressive episode, single or recurrent, were studied. Patients were assessed using a structured demographic and clinical data interview, and by the Polish versions of the HCL-32 and MDQ questionnaires. RESULTS: Hypomanic symptoms exceeding cut-off criteria for bipolarity by HCL-32 were found in 37.5% of patients and, by MDQ, in 20% of patients. Patients with HCL-32 (+) or MDQ (+) differed significantly from patients with HCl-32 (-) and MDQ (-) respectively, by being less frequently married, having more family history of depression, bipolar disorder, alcoholism and suicide, earlier onset of illness, and more depressive episodes and psychiatric hospitalizations. The percentage of patients resistant to treatment with antidepressant drugs was significantly higher in HCL-32 (+) vs. HCL-32 (-) and in MDQ (+) vs. MDQ (-): 43.9% vs. 30.0%, and 26.4% vs. 12.4%, respectively. CONCLUSIONS: The results confirm a substantial percentage of bipolarity in major depressive disorder. Such patients have a number of clinical characteristics pointing on a more severe form of the illness and their depression is more resistant to treatment with antidepressants.

Studies on microheterogeneity of acute-phase proteins in rheumatoid arthritis by using crossed affinoimmuno-electrophoresis with free concanavalin A.

Microheterogeneity of two acute-phase proteins: orosomucoid (alpha1-acid glycoprotein, AGP) and alpha 1-antichymotrypsin (ACHT) were studied in the sera of 48 patients with rheumatoid arthritis and 12 healthy individuals. Each rheumatoid patient was assigned to one of four activity grades. Cross affinoimmunoelectrophoresis (aff-EP) with free Concanavalin A (Con A) revealed three microheterogeneity variants of AGP and four microheterogeneity forms of ACHT. The relative amounts of AGP-variants and ACHT-variants observed in the healthy donors were similar to those observed in the patients with activity grade I, but they changed with the increase in the grade of rheumatoid activity. The differences were most significant for AGP. The comparison of serum C-reactive protein (CRP) levels with AGP-variant-O-non reactive with Con A showed significant correlation in respective rheumatoid activity grades.

How are alpha beta T cells positively selected in the thymus?

Thymus-derived (T) lymphocytes have the potential to express antigen receptors (TCR) that can recognize both self, as well as foreign antigens as they appear on the cell surface. In the thymus, positive selective allows the maturation of T cells that are able to see foreign antigens in conjunction with molecules encoded by genes of the major histocompatibility complex (MHC), whereas negative selection deletes auto-aggressive T cells. Control of T cell development is the only known function of the thymus. Therefore, it has been argued that recognition events responsible for selection of the T cell repertoire are guided by unique cellular interactions in the thymus. Here, we will show that positive selection can also occur on non-thymic cells. We will also argue that positive selection is not dependent on unique thymic accessory cells (AC) function. In other words, restricted recognition is not taught rather thymocytes learn it by themselves.

Thymic education--T cells do it for themselves.

The phenomenon of positive selection of developing T cells is well established. However, the cellular interactions that are responsible for selecting the restriction element are still ill defined. Here, Tomasz Pawlowski and Uwe Staerz discuss the latest developments in the field, as well as some new evidence suggesting that cells of epithelial or hematopoietic origin are not the only ones capable of selecting the restriction element for T cells expressing the alpha beta T-cell receptor.