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.

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.

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.

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.

Eccentric atherosclerotic plaques with positive remodelling have a pericardial distribution: a permissive role of epicardial fat? A three-dimensional intravascular ultrasound study of left anterior descending artery lesions.

AIMS: The transversal distribution of coronary atherosclerotic plaques (AP) (myocardial vs pericardial) affects vessel remodelling. The aim of this study was to define the impact of transversal lesion distribution on vessel remodelling in proximal and distal coronary segments using a 3D intravascular ultrasound (IVUS) reconstruction. METHODS: The study group included 70 lesions located in the left anterior descending artery within 5mm of the septal take-off, and imaged using 3D-IVUS. The take-off of the septal branch was used to divide the plaque into a myocardial and pericardial surface. The IVUS index of vessel remodelling was calculated as: [narrowest external elastic membrane (EEM) site cross-sectional area (CSA)-reference EEM CSA)/reference EEM CSAx100]. The lesions with an intermediate vessel remodelling index (between -25% and +15%) were excluded from analysis. RESULTS: Of the 38 APs with a pericardial distribution, 34 (89%) showed positive remodelling (P<0.001). The distal lesions had a positive vessel remodelling index regardless of transversal plaque distribution. At multivariate analysis, pericardial distribution and the distal location of AP were the only independent variables predictive of positive remodelling. CONCLUSIONS: The transversal distribution of atherosclerotic plaque affects vessel remodelling in left anterior descending coronary lesions, probably because of an extravascular splinting effect. Distal lesions usually show positive remodelling regardless of transversal plaque distribution.

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.

[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.

Permissive recognition during positive selection.

In the periphery alpha beta T lymphocytes recognize antigens in conjunction with major histocompatibility complex (MHC) molecules. In the thymus immature T cells are positively selected on MHC molecules in the apparent absence of cognate peptides. Thus, at different developmental stages a T cell responds to different epitopes, yet uses the identical alpha beta T cell antigen receptor (TcR). To explain this paradox it has been hypothesized that during positive selection immature T cells see peptides/ligands unique to the thymus, are selected by specific antagonists related to their cognate peptides, or are driven by lowered affinity thresholds of their TcR. Though different in detail, these theories rely on defined peptides uniquely matched to select certain TcR. However, we find that in a TcR-transgenic (TcR(trans +)) mouse severely limiting the diversity of peptides does not impair positive selection. We show that many unrelated peptides, including some naturally occurring on the cell surface, induce maturation of CD4-CD8+TcR(high) thymocytes. The same peptides when presented in conjunction with the selecting MHC molecule, are not recognized by peripheral T cells expressing the same TcR(trans). Therefore, these findings point to a promiscuous rather than discriminate recognition mode used by immature T cells.

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.

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.

[Effect of aorto-coronary bypass on function of cardiac ventricles as evaluated by non-invasive techniques during a two year observation]. / Wplyw pomostowania aortalno-wiencowego na czynnosc komór serca oceniany metodami nieinwazyjnymi w dwuletniej obserwacji.

In 1989-1990, 892 patients with ischemic heart disease were treated at the III Department of Cardiology in Katowice-Ochojec. Aortocoronary bypass was performed in 51 (5.7%) of these patients. Thirty five out of these patients were followed-up for 24 months after surgery. They reported to the hospital for evaluation after 3, 6, 12 and 14 months following a discharge. Efficiency of revascularization was checked with physical examination, and non-invasive tests such as electrocardiography, echocardiography and exercise tolerance testing. Diastolic and symbolic functioning of left and right ventricle was assessed separately. All patients suffered from anginal pain before surgery whereas 85% of operated patients were free from pain. No correlation between incomplete revascularization and recurrence of the anginal pain was noted. Surgery produced an increase in exercise tolerance and maximal load. No significant improvement in the right ventricle was seen during a 2-year follow-up. In the group of patients without previous myocardial infarction measured parameters of cardiac ventricles functioning were better than those in patients who had myocardial infarction. Therefore, the former are better candidates to coronary vessels revascularization.

Positive selection of T lymphocytes on fibroblasts.

Thymocytes are selected for expression of alpha beta T-cell antigen receptors (TCR) which recognize antigen in conjunction with self-major histocompatibility complex (MHC) molecules. In the thymus the restriction element is imprinted on radioresistant stromal elements and on cells of haematopoietic origin. In mice negative for beta 2-microglobulin that are devoid of mature cytotoxic T lymphocytes, we find that intrathymic injection of different fibroblasts causes the maturation of CD4-CD8+TCRhigh thymocytes with distinct patterns of TCR V beta distribution. Here we show that in TCR-transgenic mice, intrathymic injection of L cells expressing the selecting H-2Kb molecule (L-Kb cells) reconstitutes the maturation of thymocytes bearing the transgenic TCR, and that in normal B10.BR (H-2k) mice, H-2Kb molecules expressed on L-Kb cells lead to the development of T lymphocytes with recognition restricted to H-2Kb. A class I MHC restriction element can thus be selected by interaction with fibroblasts, that is, cells of other than epithelial or haematopoietic origin.

Different capabilities of monocytes from patients with systemic lupus erythematosus and rheumatoid arthritis to induce glycosylation alterations of acute phase proteins in vitro.

The effect of conditioned medium on the biosynthesis and glycosylation profile of acute phase proteins secreted by the human hepatoma cell line Hep G2 was studied. Conditioned medium was prepared from nonactivated [CM-LPS(-)] and ex vivo lipopolysaccharide activated [CM-LPS(+)] monocytes from eight patients with active rheumatoid arthritis (RA), five patients with active systemic lupus erythematosus (SLE), and seven healthy subjects. The biosynthesis of albumin, alpha 1-antichymotrypsin and alpha 1-proteinase inhibitor and the profile of glycosylation of proteinase inhibitor were analysed. CM-LPS(-) from patients with SLE had a similar effect to CM-LPS(-) from healthy subjects. In contrast, CM-LPS(-) from patients with RA had the same effect as CM-LPS(+) from healthy donors. A similar effect to that of CM-LPS(+) of healthy subjects was seen with CM-LPS(+) from patients with SLE and with CM-LPS(+) from patients with RA. The treatment of CM-LPS(+) with antibodies against interleukin 6 neutralised most of its ability to induce changes in the biosynthesis and glycosylation of acute phase proteins. Antibodies to interleukin 1 and tumour necrosis factor alpha had only a limited effect on the ability of CM-LPS(+) to induce changes of albumin and alpha 1-antichymotrypsin syntheses, whereas they had no effect on the biosynthesis and glycosylation of proteinase inhibitor. These results indicate that: (a) monocytes isolated from patients with active SLE and active RA have different capabilities of inducing alterations of acute phase proteins in vitro; (b) ex vivo activation of monocytes from patients with SLE leads to the full induction of its capabilities to change acute phase proteins, whereas the activation of monocytes from patients with RA has no additive effects; and (c) interleukin 6 seems to be a major cytokine involved in the regulation of the glycosylation pattern of acute phase proteins.

[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.

Toxoplasmosis and systemic lupus erythematosus. Comparative serological studies.

There are still controversial views as to the relation between SLE and toxoplasmosis. Therefore, we looked for serological markers in both diseases. In patients with SLE (17), toxoplasmosis (28), and in normal controls (28) anti-Toxoplasma gondii antibodies, anti-nuclear antibodies of different specificities, anti-histone and anti-cardiolipine antibodies, as well as antibodies against most common public idiotypes were measured. Significant increases in antinuclear antibodies and other SLE-related antigens were observed in patients with SLE. On the contrary, low levels of these antibodies were found in toxoplasmosis patients and in controls. The same was true for 16/6 anti-DNA idiotype antibodies. The incidence of anti-Toxoplasma antibodies in SLE sera did not differ in comparison with that in the normal population. Our data suggest that subacute and chronic toxoplasmosis do not play essential roles in generating the antibodies that are important to the pathogenetic mechanism operating in SLE.

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.