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1.
Angiogenin and vascular endothelial growth factor expression in lungs of lung cancer patients
Aleš Rozman, Mira Šilar, Mitja Košnik, 2012, izvirni znanstveni članek

Povzetek: Background. Lung cancer is the leading cause of cancer deaths. Angiogenesis iscrucial process in cancer growth and progression. This prospective study evaluated expression of two central regulatory molecules: angiogenin and vascular endothelial growth factor (VEGF) in patients with lung cancer. Patients and methods. Clinical data, blood samples and broncho-alveolar lavage(BAL) from 23 patients with primary lung carcinoma were collected. BAL fluid was taken from part of the lung with malignancy, and from corresponding healthy side of the lung. VEGF and angiogenin concentrations were analysed by an enzyme-linked immunosorbent assay. Dilution of bronchial secretions in the BAL fluid was calculated from urea concentration ratio between serum and BAL fluid. Results. We found no statistical correlation between angiogenin concentrations in serum and in bronchial secretions from both parts of the lung. VEGF concentrations were greater in bronchial secretions in the affectedside of the lung than on healthy side. Both concentrations were greater than serum VEGF concentration. VEGF concentration in serum was in positive correlation with tumour size (p = 0,003) and with metastatic stage ofdisease (p = 0,041). There was correlation between VEGF and angiogenin concentrations in bronchial secretions from healthy side of the lung and between VEGF and angiogenin concentrations in bronchial secretions from part of the lung with malignancy. Conclusion. Angiogenin and VEGF concentrations insystemic, background and local samples of patients with lung cancer are affected by different mechanisms. Pro-angiogenic activity of lung cancer has an important influence on the levels of angiogenin and VEGF.
Objavljeno v DiRROS: 22.03.2024; Ogledov: 34; Prenosov: 17
.pdf Celotno besedilo (467,61 KB)

2.
Comparison of colorimetric MTT and clonogenic assays for irradiation and cisplatin treatment on murine fibrosarcoma SA-1 cells
Maja Čemažar, Darja Marolt, Mira Lavrič, Gregor Serša, 1999, izvirni znanstveni članek

Povzetek: Background. The aim of our study was to determine the relationship between cell survival of SA-1 tumor cells measured by clonogenic assay and MTT assay after irradiation and cisplatin treatment. Materials and methods. Survival of SA-1 cells was measured after irradiation (2-8 Gy) and cisplatin treatment (0.05-0.5 u g/ml) by clonogenic assay performed 7 days after treatment, and byMTT assay performed on day 3, 4, 5, and 7 after the treatment. Results. The results showed good correlation between MTT assay and clonogenic assay for irradiation doses below 4 Gy. For higher doses good correlation between MTT and clonogenic assay was determined only when MTT assay was performed on day 5and 7 after the treatment. In the case of cisplatin treatment, similar pattern was observed, good correlation between IC50 values for MTT and clonogenic assay was found when MTT assay was performed on day 5 and 7 after the treatment. Conclusion. Results of our study confirmed the results of previous studies addressing this topic and further support the use of MTT testas an alternative test for clonogenic test as a predictive assay of tumourresponse to the radio or chemotherapy.
Objavljeno v DiRROS: 22.01.2024; Ogledov: 151; Prenosov: 33
.pdf Celotno besedilo (419,17 KB)

3.
Reduced parasympathetic reactivation during recovery from exercise in myalgic encephalomyelitis/chronic fatigue syndrome
Jessica Van Oosterwijck, Uroš Marušič, Inge De Wandele, Mira Meeus, Lorna Paul, Luc Lambrecht, Greta Moorkens, Lieven Danneels, Jo Nijs, 2021, izvirni znanstveni članek

Povzetek: Although autonomic nervous system (ANS) dysfunction in Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) has been proposed, conflicting evidence makes it difficult to draw firm conclusions regarding ANS activity at rest in ME/CFS patients. Although severe exercise intolerance is one of the core features of ME/CFS, little attempts have been made to study ANS responses to physical exercise. Therefore, impairments in ANS activation at rest and following exercise were examined using a case-control study in 20 ME/CFS patients and 20 healthy people. Different autonomous variables, including cardiac, respiratory, and electrodermal responses were assessed at rest and following an acute exercise bout. At rest, parameters in the time-domain represented normal autonomic function in ME/CFS, while frequency-domain parameters indicated the possible presence of diminished (para)sympathetic activation. Reduced parasympathetic reactivation during recovery from exercise was observed in ME/CFS. This is the first study showing reduced parasympathetic reactivation during recovery from physical exercise in ME/CFS. Delayed HR recovery and/or a reduced HRV as seen in ME/CFS have been associated with poor disease prognosis, high risk for adverse cardiac events, and morbidity in other pathologies, implying that future studies should examine whether this is also the case in ME/CFS and how to safely improve HR recovery in this population.
Ključne besede: autonomic nervous system, autonomic function, electrodermal activity, electrocardiogram, heart rate, physical activity
Objavljeno v DiRROS: 20.10.2021; Ogledov: 838; Prenosov: 720
.pdf Celotno besedilo (4,50 MB)
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Zdravstvena nega onkološkega bolnika, ki ima bolečine
Mira Logonder, 1997, objavljeni strokovni prispevek na konferenci

Objavljeno v DiRROS: 11.05.2021; Ogledov: 720; Prenosov: 166
.pdf Celotno besedilo (839,08 KB)

6.
Mast cell activation test in the diagnosis of allergic disease and anaphylaxis
Rajia Bahri, Adnan Custovic, Peter Korošec, Marina Tsoumani, Martin Barron, Jiakai Wu, Rebekah Sayers, Alf Weimann, Monica Ruiz-Garcia, Nandinee Patel, Mira Šilar, 2018, izvirni znanstveni članek

Povzetek: Background. Food allergy is an increasing public health issue and the commonest cause of life-threatening anaphylactic reactions. Conventional allergy tests assess for the presence of allergen-specific IgE, significantly overestimating the rate of true clinical allergy resulting in over-diagnosis and adverse impact on health-related quality of life. Objective. To undertake initial validation and assessment of a novel diagnostic tool, the mast cell activation test (MAT). Methods. Primary human mast cells (hMCs) were generated from peripheral blood precursors, and sensitized using patient sera and then incubated with allergen. Mast cell degranulation was assessed by flow cytometry and mediator release. We compared the diagnostic performance of MAT to existing diagnostic tools to assess in a cohort of peanut-sensitized individuals undergoing double-blind, placebo-controlled challenge. Results. hMCs sensitized with sera from peanut, grass pollen and hymenoptera- (wasp venom) allergic patients demonstrated allergen-specific and dose-dependent degranulation by both expression of surface activation markers (CD63 and CD107a) and functional assays (prostaglandins D2 and ß-hexosaminidase release). In this cohort of peanut-sensitized individuals, MAT was found to have superior discrimination performance compared to other testing modalities including component-resolved diagnostics and basophil activation test. Using functional principle component analysis, we identified 5 clusters or patterns of reactivity in the resulting dose-response curves, which at preliminary analysis corresponded to the reaction phenotypes seen at challenge. Conclusion. MAT is a robust tool which may confer superior diagnostic performance compared to existing allergy diagnostics, and may be useful to explore differences in effector cell function between basophils and mast cells during allergic reactions.
Ključne besede: allergy and immunology -- diagnosis, anaphylaxis, immunologic tests, mast cells, food hypersensitivity, basophil activation test, BAT, mast cell activation test
Objavljeno v DiRROS: 30.11.2020; Ogledov: 1644; Prenosov: 1555
.pdf Celotno besedilo (3,30 MB)
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7.
Systemic and airway oxidative stress in competitive swimmers
Sabina Škrgat, Peter Korošec, Izidor Kern, Mira Šilar, Julij Šelb, Matjaž Fležar, Robert Marčun, 2018, izvirni znanstveni članek

Povzetek: Background: The environment in swimming pools, which contain chlorine, might interact with the airway epithelium, resulting in oxidative stress and/or inflammation during high intensity training periods. Methods: We evaluated pulmonary functional (metacholine challenge test, FEV1 and VC), cellular (eosinophils and neutrophils), inflammatory (FeNo, IL-5, IL-6, IL-8 and TNF-[alpha]), oxidative (8-isoprostanes) and angiogenesis factors (VEGF) in induced sputum and peripheral blood of 41 healthy non-asthmatic elite swimmers (median 16 years) during the period of high intensity training before a national championship. The second paired sampling was performed seven months later after training had been stopped for one month. Results: There was a ten-fold increase (median 82-924 pg/ml; P < 0.001) in 8-isoprostanes in induced sputum and five-fold increase (median 82-924 pg/ml; P < 0.001) in sera during training in comparison to the period of rest. However, there was no difference in FEV1 (113 vs 116%), VC (119 vs 118%), FeNo (median 34 vs 38 ppb), eosinophils (2.7 vs 2.9% in sputum; 180 vs 165 cells/[micro]l in blood), neutrophils, different cytokines or VEGF in induced sputum or sera. The only exception was TNF-[alpha], which was moderately increased in sera (median 23 vs 40 pg/ml; P=0.02) during the peak training period. Almost half (18 of 41) of swimmers showed bronchial hyperresponsiveness during the peak training period (PC20 cutoff was 4 mg/ml). There was no correlation between hyperresponsiveness and the markers of oxidative stress or inflammation. Conclusions: High intensity training in healthy, non-asthmatic competitive swimmers results in marked oxidative stress at the airway and systemic levels, but does not lead to airway inflammation. However, we could not confirm that oxidative stress is associated with bronchial hyperresponsiveness (AHR), which is often observed during the peak exercise training period.
Ključne besede: bronchial diseases, swimming, oxidative stress, bronchial hyperresponsiveness, competitive swimmers, training
Objavljeno v DiRROS: 23.11.2020; Ogledov: 1638; Prenosov: 451
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8.
The culprit insect but not severity of allergic reactions to bee and wasp venom can be determined by molecular diagnosis
Pia Gattinger, Christian Lupinek, Lampros Kalogiros, Mira Šilar, Mihaela Zidarn, Peter Korošec, Christine Koessler, Natalija Novak, Rudolf Valenta, Irene Mittermann, 2018, izvirni znanstveni članek

Povzetek: Background. Allergy to bee and wasp venom can lead to life-threatening systemic reactions. The identification of the culprit species is important for allergen-specific immunotherapy. Objectives. To determine a panel of recombinant bee and wasp allergens which is suitable for the identification of bee or wasp as culprit allergen sources and to search for molecular surrogates of clinical severity of sting reactions. Methods. Sera from eighty-seven patients with a detailed documentation of their severity of sting reaction (Mueller grade) and who had been subjected to titrated skin testing with bee and wasp venom were analyzed for bee and wasp-specific IgE levels by ImmunoCAPTM. IgE-reactivity testing was performed using a comprehensive panel of recombinant bee and wasp venom allergens (rApi m 1, 2, 3, 4, 5 and 10; rVes v 1 and 5) by ISAC chip technology, ImmunoCAP and ELISA. IgG4 antibodies to rApi m 1 and rVes v 5 were determined by ELISA and IgE/ IgG4 ratios were calculated. Results from skin testing, IgE serology and IgE/IgG4 ratios were compared with severity of sting reactions. Results. The panel of rApi m 1, rApi m 10, rVes v 1 and rVes v 5 allowed identification of the culprit venom in all but two of the 87 patients with good agreement to skin testing. Severities of sting reactions were not associated with results obtained by skin testing, venom-specific IgE levels or molecular diagnosis. Severe sting reactions were observed in patients showing < 1 ISU and < 2kUA/L of IgE to Api m 1 and/or Ves v 5. Conclusion. We identified a minimal panel of recombinant bee and wasp allergens for molecular diagnosis which may permit identification of bee and/or wasp as culprit insect in venom-sensitized subjects. The severity of sting reactions was not associated with parameters obtained by molecular diagnosis.
Ključne besede: allergy and immunology -- diagnosis, allergens -- diagnosis, hymenoptera, immunotherapy, bee, wasp, venom, sting reactions, molecular diagnosis, systemic reactions
Objavljeno v DiRROS: 23.11.2020; Ogledov: 12464; Prenosov: 1530
.pdf Celotno besedilo (1,20 MB)
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9.
Quantitative, absolute count-based T cell analysis of CD69 upregulation as a new methodology for in vitro diagnosis of delayed-type nickel hypersensitivity
Ana Koren, Mira Šilar, Helena Rupnik, Mihaela Zidarn, Peter Korošec, 2019, izvirni znanstveni članek

Povzetek: Background: T cells play a major role in delayed-type hypersensitivity reactions. Their reactivity can be assessed by measuring the upregulation of the activation marker CD69, followed by proliferation and cytokine production. The aim of our study was to develop a novel, whole blood-based, quantitative, absolute count activation index (AI) analysis of CD69 upregulation on different subsets of T cells in nickel hypersensitive patients and compare it with the previously reported approaches. Methods: Ten patients with nickel allergy and nine healthy controls were included. CD69 expression of CD3+, CD3+CD4+ and CD3+CD8+ T cells in heparinized blood was determined with flow cytometry after incubation with nickel sulfate for 48 h. The absolute cell count of CD69+ cells was determined with microbeads. The production of the cytokines IL-2, IL-5, IL-13, and IFN-[gamma] was determined after nickel sulfate stimulation of PBMNCs for 48 h. Results: We showed that the most sensitive methodology is the absolute AI, which was calculated as the ratio between the absolute count of CD69-positive T cells stimulated with nickel and the absolute count of CD69-positive T cells in non-stimulated blood. This novel quantitative approach was more discriminative than the previously reported approaches in which T cell CD69 percentage AI and cytokine production are measured. Conclusions: Our results demonstrated that measuring the absolute CD69 AI is an accurate new approach to quantify antigen-specific T cells in the blood of patients with hypersensitivity reactions to nickel. This approach may be useful for better in vitro assessment of patients with delayed-type hypersensitivity reactions.
Ključne besede: allergy and immunology, delayed hypresensitivity, nickel, blood, CD antigenes, CD69
Objavljeno v DiRROS: 18.11.2020; Ogledov: 1158; Prenosov: 296
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10.
CD3+CD4-CD8- mucosal T cells are associated with uncontrolled chronic rhinosinusitis with nasal polyps
Tanja Soklič, Mira Šilar, Matija Rijavec, Ana Koren, Izidor Kern, Irena Hočevar-Boltežar, Peter Korošec, 2019, kratki znanstveni prispevek

Povzetek: Increased mucosal double-negative (DN) CD3+CD4-CD8- T cells were found for the first time in CRS and were much more abundant in uncontrolled CRSwNP than in well-controlled CRSwNP.
Ključne besede: chronic rhinosinusitis, CD3+ T-cells, CD4- T-cells, CD8- T-cells
Objavljeno v DiRROS: 22.10.2020; Ogledov: 1516; Prenosov: 496
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