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Article ; Online: Current status of the lateral flow immunoassay for the detection of SARS-CoV-2 in nasopharyngeal swabs.

Somborac Bačura, Anita / Dorotić, Marija / Grošić, Leonarda / Džimbeg, Monika / Dodig, Slavica

2021 Volume 31, Issue 2, Page(s) 20601

Abstract: Early detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and diagnosis of coronavirus disease 2019 (COVID-19) are priorities during the pandemic. Symptomatic and suspected asymptomatic individuals should be tested for COVID-19 to ... ...

Abstract	Early detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and diagnosis of coronavirus disease 2019 (COVID-19) are priorities during the pandemic. Symptomatic and suspected asymptomatic individuals should be tested for COVID-19 to confirm infection and to be excluded from social interactions. As molecular testing capacity is overloaded during the pandemic, rapid antigen tests, such as lateral flow immunoassays (LFIAs), can be a useful tool as they allow greater test availability and obtain results in a very short time. This short review aims to present the analytical properties of LFIAs in the detection of SARS-CoV-2 in nasopharyngeal swabs. Lateral flow immunoassay is a method that combines thin-layer chromatography and indirect immunochemical sandwich method and allows the detection of a specific SARS-CoV-2 antigen in nasopharyngeal swabs. Swab specimens should be adequately collected and tested as soon as possible. Users should pay attention to quality control and possible interferences. Antigen tests for SARS-CoV-2 show high sensitivity and specificity in cases with high viral loads, and should be used up to five days after the onset of the first symptoms of COVID-19. False positive results may be obtained when screening large populations with a low prevalence of COVID-19 infection, while false negative results may happen due to improper specimen collection or insufficient amount of antigen in the specimen. So as to achieve reliable results, a diagnostic accuracy study of a specific rapid antigen test should be performed.
MeSH term(s)	Antigens, Viral/analysis ; COVID-19/diagnosis ; COVID-19/virology ; False Negative Reactions ; Humans ; Immunoassay/methods ; Immunoassay/standards ; Limit of Detection ; Nasopharynx/virology ; Point-of-Care Systems ; Quality Control ; SARS-CoV-2/immunology ; SARS-CoV-2/isolation & purification ; SARS-CoV-2/metabolism ; Sensitivity and Specificity ; Viral Load
Chemical Substances	Antigens, Viral
Language	English
Publishing date	2021-06-10
Publishing country	Croatia
Document type	Journal Article ; Review
ZDB-ID	1208725-7
ISSN	1846-7482 ; 1330-0962
ISSN (online)	1846-7482
ISSN	1330-0962
DOI	10.11613/BM.2021.020601
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Article ; Online: Extracellular Hsp70 modulates 16HBE cells' inflammatory responses to cigarette smoke and bacterial components lipopolysaccharide and lipoteichoic acid.

Hulina-Tomašković, Andrea / Somborac-Bačura, Anita / Grdić Rajković, Marija / Hlapčić, Iva / Jonker, Marnix R / Heijink, Irene H / Rumora, Lada

Cell stress & chaperones

2022 Volume 27, Issue 5, Page(s) 587–597

Abstract: Cigarette smoke is a major risk factor for chronic obstructive pulmonary disease (COPD), leading to chronic inflammation, while bacterial components lipopolysaccharide (LPS) and lipoteichoic acid (LTA) are often present in airways of COPD patients, ... ...

Abstract	Cigarette smoke is a major risk factor for chronic obstructive pulmonary disease (COPD), leading to chronic inflammation, while bacterial components lipopolysaccharide (LPS) and lipoteichoic acid (LTA) are often present in airways of COPD patients, especially during exacerbations.We hypothesised that extracellular heat shock protein 70 (eHsp70), a damage-associated molecular pattern elevated in serum of COPD patients, induces inflammation and alters cigarette smoke and LPS/LTA-induced inflammatory effects in the airway epithelium.We used 16HBE cells exposed to recombinant human (rh)Hsp70 and its combinations with cigarette smoke extract (CSE), LPS or LTA to investigate those assumptions, and we determined pro-inflammatory cytokines' secretion as well as TLR2 and TLR4 gene expression.rhHsp70 and CSE alone stimulated IL-6, IL-8 and TNF-α secretion. CSE and rhHsp70 had antagonistic effect on IL-6 secretion, while combinations of LPS or LTA with rhHsp70 showed antagonistic effect on TNF-α release. By using specific inhibitors, we demonstrated that effects of rhHsp70 on cytokines' secretion were mediated via NF-κB and/or MAPK signalling pathways. rhHsp70 increased, and CSE decreased TLR2 gene expression compared to untreated cells, but their combinations increased it compared to CSE alone. LPS and rhHsp70 combinations decreased TLR2 gene expression compared to untreated cells. TLR4 expression was not induced by any of the treatments.In conclusion, we demonstrated that extracellular Hsp70 modulates pro-inflammatory responses of human airway epithelial cells to cigarette smoke and bacterial components LPS and LTA. Simultaneous presence of those compounds and their interactions might lead to inappropriate immune responses and adverse consequences in COPD.
MeSH term(s)	Cigarette Smoking ; HSP70 Heat-Shock Proteins/genetics ; HSP70 Heat-Shock Proteins/metabolism ; Humans ; Inflammation/metabolism ; Interleukin-6 ; Interleukin-8 ; Lipopolysaccharides/pharmacology ; NF-kappa B/metabolism ; Pulmonary Disease, Chronic Obstructive/metabolism ; Teichoic Acids ; Nicotiana/adverse effects ; Nicotiana/metabolism ; Toll-Like Receptor 2/genetics ; Toll-Like Receptor 2/metabolism ; Toll-Like Receptor 4/genetics ; Toll-Like Receptor 4/metabolism ; Tumor Necrosis Factor-alpha
Chemical Substances	HSP70 Heat-Shock Proteins ; Interleukin-6 ; Interleukin-8 ; Lipopolysaccharides ; NF-kappa B ; Teichoic Acids ; Toll-Like Receptor 2 ; Toll-Like Receptor 4 ; Tumor Necrosis Factor-alpha ; lipoteichoic acid (56411-57-5)
Language	English
Publishing date	2022-08-27
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1362749-1
ISSN	1466-1268 ; 1355-8145
ISSN (online)	1466-1268
ISSN	1355-8145
DOI	10.1007/s12192-022-01294-w
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Article ; Online: Cigarette smoke and extracellular Hsp70 induce secretion of ATP and differential activation of NLRP3 inflammasome in monocytic and bronchial epithelial cells.

Rumora, Lada / Somborac-Bačura, Anita / Hlapčić, Iva / Hulina-Tomašković, Andrea / Rajković, Marija Grdić

Cytokine

2020 Volume 135, Page(s) 155220

Abstract: Background: Chronic obstructive pulmonary disease (COPD) is an inflammatory disease mainly caused by smoking. Cigarette smoke damages airway epithelium and activates lung macrophages, causing inflammatory responses. It was suggested that nucleotide- ... ...

Abstract	Background: Chronic obstructive pulmonary disease (COPD) is an inflammatory disease mainly caused by smoking. Cigarette smoke damages airway epithelium and activates lung macrophages, causing inflammatory responses. It was suggested that nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome might have an important role in COPD development. Study aimed to explore whether cigarette smoke extract (CSE), extracellular heat shock protein 70 (eHsp70) or their combinations induce adenosine triphosphate (ATP) release and NLRP3 inflammasome activation. Methods: We detected NLRP3 and interleukin (IL)-1β mRNA expression, extracellular IL-1β and ATP concentrations as well as lactate dehydrogenase (LDH) activity. We used bronchial epithelial (NCI-H292, 16HBE and NHBE) and monocytic cells (monocyte-derived macrophages (MDMs) and THP-1) as representative of local airway and systemic compartments that could be affected in COPD. Results: CSE and eHsp70 increased NLRP3 and IL-1β mRNA expression as well as IL-1β and ATP secretion in all cells compared to untreated cells. Lytic cell death was observed in cell lines, especially those of bronchial epithelium origin, but not in primary cells (NHBE, MDMs). Regarding LDH activity, eHsp70 did not modulate CSE effects, except in NCI-H292 cell line. However, eHsp70 significantly affected CSE-provoked NLRP3 inflammasome activation by causing mostly antagonistic effects in airway epithelial cells and synergistic effects in MDMs. Conclusion: We demonstrated that both CSE and eHsp70 induce ATP secretion and differential activation of NLRP3 inflammasome in bronchial epithelial and monocytic cells. We suggest that these mechanisms might be involved in pathophysiology of COPD by contributing to the propagation of inflammation.
MeSH term(s)	Adenosine Triphosphate/metabolism ; Bronchi/drug effects ; Bronchi/metabolism ; Cell Line ; Epithelial Cells/drug effects ; Epithelial Cells/metabolism ; HSP70 Heat-Shock Proteins/metabolism ; Humans ; Inflammasomes/drug effects ; Inflammasomes/metabolism ; Inflammation/metabolism ; Interleukin-1beta/metabolism ; L-Lactate Dehydrogenase/metabolism ; Macrophages/drug effects ; Macrophages/metabolism ; Monocytes/metabolism ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism ; Pulmonary Disease, Chronic Obstructive/metabolism ; Smoke/adverse effects ; Smoking/adverse effects ; Smoking/metabolism ; Nicotiana/adverse effects
Chemical Substances	HSP70 Heat-Shock Proteins ; Inflammasomes ; Interleukin-1beta ; NLR Family, Pyrin Domain-Containing 3 Protein ; NLRP3 protein, human ; Smoke ; Adenosine Triphosphate (8L70Q75FXE) ; L-Lactate Dehydrogenase (EC 1.1.1.27)
Language	English
Publishing date	2020-07-28
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1018055-2
ISSN	1096-0023 ; 1043-4666
ISSN (online)	1096-0023
ISSN	1043-4666
DOI	10.1016/j.cyto.2020.155220
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Article ; Online: Pathogen-associated molecular patterns and extracellular Hsp70 interplay in NLRP3 inflammasome activation in monocytic and bronchial epithelial cellular models of COPD exacerbations.

Rumora, Lada / Hlapčić, Iva / Hulina-Tomašković, Andrea / Somborac-Bačura, Anita / Bosnar, Martina / Rajković, Marija Grdić

APMIS : acta pathologica, microbiologica, et immunologica Scandinavica

2020 Volume 129, Issue 2, Page(s) 80–90

Abstract: During chronic obstructive pulmonary disease (COPD) exacerbations, interplay between pathogen-associated molecular patterns (PAMPs; e.g. lipopolysaccharide (LPS) and lipoteichoic acid (LTA)) and damage-associated molecular patterns (DAMPs; e.g. ... ...

Abstract	During chronic obstructive pulmonary disease (COPD) exacerbations, interplay between pathogen-associated molecular patterns (PAMPs; e.g. lipopolysaccharide (LPS) and lipoteichoic acid (LTA)) and damage-associated molecular patterns (DAMPs; e.g. extracellular heat shock protein 70 (eHsp70) and adenosine triphosphate (ATP)) might influence patient's outcome. Nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome might have a role in dysfunctional immune system in COPD. We hypothesized that LPS, LTA, eHsp70 and their combinations induce NLRP3 inflammasome activation, and we aimed to explore this assumption. We used monocytic (monocyte-derived macrophages (MDMs) and THP-1) and bronchial epithelial cells (NHBE and NCI-H292) to represent systemic and local airway compartments that could be affected in COPD. Bacterial components and eHsp70 stimulated NLRP3 and interleukin (IL)-1β gene expression as well as IL-1β and ATP release from all cells compared to non-treated cells. LDH secretion was induced in cell lines only. eHsp70 had inhibitory (NCI-H292) or stimulatory (NHBE) effects on eATP levels compared to PAMP alone. Regarding NLRP3 inflammasome activation, eHsp70 had mostly antagonistic effects. We demonstrated that bacterial components and eHsp70 activate NLRP3 inflammasome and increase ATP secretion. We suggest that extracellular Hsp70 might modulate immune responses provoked by bacterial infections and affect COPD patients' outcome during acute exacerbations.
MeSH term(s)	Adenosine Triphosphate/metabolism ; Alveolar Epithelial Cells/metabolism ; Alveolar Epithelial Cells/pathology ; Cell Line ; HSP70 Heat-Shock Proteins/metabolism ; Humans ; Interleukin-1beta/metabolism ; Lipopolysaccharides/metabolism ; Macrophages/metabolism ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism ; Pathogen-Associated Molecular Pattern Molecules/metabolism ; Pulmonary Disease, Chronic Obstructive/pathology ; THP-1 Cells ; Teichoic Acids/metabolism
Chemical Substances	HSP70 Heat-Shock Proteins ; Hspa14 protein, human ; IL1B protein, human ; Interleukin-1beta ; Lipopolysaccharides ; NLR Family, Pyrin Domain-Containing 3 Protein ; NLRP3 protein, human ; Pathogen-Associated Molecular Pattern Molecules ; Teichoic Acids ; lipoteichoic acid (56411-57-5) ; Adenosine Triphosphate (8L70Q75FXE)
Language	English
Publishing date	2020-10-28
Publishing country	Denmark
Document type	Journal Article
ZDB-ID	93340-5
ISSN	1600-0463 ; 0903-4641
ISSN (online)	1600-0463
ISSN	0903-4641
DOI	10.1111/apm.13089
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Article ; Online: Circulating Tumor Cells in Colorectal Cancer: Detection Systems and Clinical Utility.

Petrik, József / Verbanac, Donatella / Fabijanec, Marija / Hulina-Tomašković, Andrea / Čeri, Andrea / Somborac-Bačura, Anita / Petlevski, Roberta / Grdić Rajković, Marija / Rumora, Lada / Krušlin, Božo / Štefanović, Mario / Ljubičić, Neven / Baršić, Neven / Hanžek, Antonija / Bočkor, Luka / Ćelap, Ivana / Demirović, Alma / Barišić, Karmela

International journal of molecular sciences

2022 Volume 23, Issue 21

Abstract: Colorectal cancer (CRC) is the third most common cancer worldwide. The high mortality from CRC is mainly related to metastasis affecting distant organs and their function. Dissemination of tumor cells from the primary tumor and hematogeneous spread are ... ...

Abstract	Colorectal cancer (CRC) is the third most common cancer worldwide. The high mortality from CRC is mainly related to metastasis affecting distant organs and their function. Dissemination of tumor cells from the primary tumor and hematogeneous spread are considered crucial in the formation of tumor metastases. The analysis of circulating tumor cells (CTCs) and CTC clusters in the blood can be used for the early detection of invasive cancer. Moreover, CTCs have a prognostic significance in the monitoring of a malignant disease or the response to chemotherapy. This work presents an overview of the research conducted on CTCs with the aim of finding suitable detection systems and assessing the possibility of clinical applications in patients with CRC.
MeSH term(s)	Humans ; Neoplastic Cells, Circulating/pathology ; Cell Count ; Colorectal Neoplasms/diagnosis ; Colorectal Neoplasms/pathology ; Biomarkers, Tumor
Chemical Substances	Biomarkers, Tumor
Language	English
Publishing date	2022-11-05
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms232113582
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Article ; Online: Extracellular adenosine triphosphate is associated with airflow limitation severity and symptoms burden in patients with chronic obstructive pulmonary disease.

Hlapčić, Iva / Hulina-Tomašković, Andrea / Somborac-Bačura, Anita / Rajković, Marija Grdić / Dugac, Andrea Vukić / Popović-Grle, Sanja / Rumora, Lada

Scientific reports

2019 Volume 9, Issue 1, Page(s) 15349

Abstract: Extracellular adenosine triphosphate (eATP)-driven inflammation was observed in chronic obstructive pulmonary disease (COPD) but was not investigated in patients' blood. Therefore, this study aimed to investigate eATP concentration in plasma of COPD ... ...

Abstract	Extracellular adenosine triphosphate (eATP)-driven inflammation was observed in chronic obstructive pulmonary disease (COPD) but was not investigated in patients' blood. Therefore, this study aimed to investigate eATP concentration in plasma of COPD patients and its association with disease severity and smoking. Study included 137 patients with stable COPD and 95 control subjects. eATP concentration was determined in EDTA plasma by luminometric method, and mRNA expression of eATP receptors P2X7R and P2Y2R was analysed by quantitative polymerase chain reaction (qPCR). eATP concentration was increased in COPD patients compared to controls (P < 0.001). Moreover, it was increasing with disease severity (GOLD 2-4) as well as symptoms burden and exacerbations history (GOLD A-D) (P < 0.05). eATP in healthy smokers differed from healthy non-smokers (P < 0.05) but was similar to GOLD 2 and GOLD A patients. eATP showed great diagnostic performances (OR = 12.98, P < 0.001) and correctly classified 79% of study participants. It demonstrated association with FEV
MeSH term(s)	Adenosine Triphosphate/metabolism ; Aged ; Aged, 80 and over ; Extracellular Space/metabolism ; Female ; Gene Expression Regulation ; Humans ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive/metabolism ; Pulmonary Disease, Chronic Obstructive/pathology ; Pulmonary Disease, Chronic Obstructive/physiopathology ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Receptors, Purinergic P2/genetics ; Receptors, Purinergic P2/metabolism ; Respiratory Function Tests ; Severity of Illness Index ; Smoking/adverse effects
Chemical Substances	RNA, Messenger ; Receptors, Purinergic P2 ; Adenosine Triphosphate (8L70Q75FXE)
Language	English
Publishing date	2019-10-25
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-019-51855-w
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Article ; Online: Platelet indices in stable chronic obstructive pulmonary disease - association with inflammatory markers, comorbidities and therapy.

Hlapčić, Iva / Somborac-Bačura, Anita / Popović-Grle, Sanja / Vukić Dugac, Andrea / Rogić, Dunja / Rako, Ivana / Žanić Grubišić, Tihana / Rumora, Lada

Biochemia medica

2019 Volume 30, Issue 1, Page(s) 10701

Abstract: Introduction: Chronic obstructive pulmonary disease (COPD) is a complex inflammatory condition that can affect haemostasis. This study aimed to determine differences in platelet-related parameters between controls and COPD subjects. The hypothesis was ... ...

Abstract	Introduction: Chronic obstructive pulmonary disease (COPD) is a complex inflammatory condition that can affect haemostasis. This study aimed to determine differences in platelet-related parameters between controls and COPD subjects. The hypothesis was that platelet indices are disturbed in COPD patients, and this would be accompanied by increased C-reactive protein (CRP), fibrinogen (Fbg) and white blood cells (WBC). Therefore, platelet count (Plt), platelet-related parameters - mean platelet volume (MPV), platelet distribution width (PDW), plateletcrit (Pct), their ratios (MPV/Plt, MPV/Pct, PDW/Plt, PDW/Pct), platelet to lymphocyte ratio (PLR), Plt index as well as CRP, Fbg and WBC were assessed. Materials and methods: Study included 109 patients with stable COPD and 95 control subjects, recruited at Clinical Department for Lung Diseases Jordanovac, University Hospital Centre Zagreb (Zagreb, Croatia). Complete blood count was performed on Sysmex XN-1000, CRP on Cobas c501, and Fbg on BCS XP analyser. Data were analysed with MedCalc statistical software. Results: Platelet (P = 0.007) and PLR (P = 0.006) were increased, while other platelet indices were decreased in COPD patients compared to controls. Combined model that included PLR, PDW and WBC showed great diagnostic performances, and correctly classified 75% of cases with an AUC of 0.845 (0.788 - 0.892), P < 0.001. Comorbidities (cardiovascular or metabolic diseases) had no effect on investigated parameters, while inhaled corticosteroids/long-acting β Conclusion: Platelet indices were altered in COPD patients and they could be valuable as diagnostic markers of COPD development, especially if combined with already known inflammatory markers.
MeSH term(s)	Adrenal Cortex Hormones/therapeutic use ; Adrenergic beta-2 Receptor Agonists/therapeutic use ; Aged ; Aged, 80 and over ; Biomarkers/blood ; Blood Platelets/cytology ; Blood Platelets/metabolism ; Cardiovascular Diseases/complications ; Cardiovascular Diseases/pathology ; Case-Control Studies ; Female ; Forced Expiratory Volume ; Humans ; Leukocytes/cytology ; Logistic Models ; Lymphocytes/cytology ; Male ; Metabolic Diseases/complications ; Metabolic Diseases/pathology ; Middle Aged ; Pulmonary Disease, Chronic Obstructive/drug therapy ; Pulmonary Disease, Chronic Obstructive/pathology
Chemical Substances	Adrenal Cortex Hormones ; Adrenergic beta-2 Receptor Agonists ; Biomarkers
Language	English
Publishing date	2019-12-15
Publishing country	Croatia
Document type	Journal Article
ZDB-ID	1208725-7
ISSN	1846-7482 ; 1330-0962
ISSN (online)	1846-7482
ISSN	1330-0962
DOI	10.11613/BM.2020.010701
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Article ; Online: Cigarette Smoke Induces Activation of Polymorphonuclear Leukocytes.

Somborac-Bačura, Anita / Popović-Grle, Sanja / Zovko, Vlasta / Žanić-Grubišić, Tihana

Lung

2017 Volume 196, Issue 1, Page(s) 27–31

Abstract: Introduction: Cigarette smoking is a major risk factor for chronic obstructive pulmonary disease (COPD). Exposure to cigarette smoke may stimulate inflammatory response and activate polymorphonuclear leukocytes (PMN) thus resulting in secretion of ... ...

Abstract	Introduction: Cigarette smoking is a major risk factor for chronic obstructive pulmonary disease (COPD). Exposure to cigarette smoke may stimulate inflammatory response and activate polymorphonuclear leukocytes (PMN) thus resulting in secretion of cellular proteases. The aim of our study was to explore the effect of cigarette smoke extract (CSE) on the release of matrix metalloproteinase-9 (MMP-9) from PMN. Methods: The study included 23 patients with stable COPD and 9 healthy controls. PMN were isolated from blood of all participants and exposed to 4% CSE or basal culture medium (0% CSE) for 20 h. MMP-9 concentration in PMN culture media was measured using the ELISA method. Results: Exposure of PMN to 4% CSE did not cause cytotoxic effects, as determined by no changes in lactate dehydrogenase (LDH) activity in PMN culture media when compared to untreated PMN (P = 0.689). In basal conditions, PMN of COPD patients released significantly more MMP-9 compared with PMN of healthy controls (P = 0.016). However, concentration ratio of MMP-9 released from PMN exposed to 4% CSE or 0% CSE of each participant was significantly higher for healthy subjects than for COPD patients (P = 0.025). Conclusion: Cigarette smoke induces activation of PMN in healthy controls. However, chronically activated PMN in COPD patients could not be further stimulated by in vitro exposure to CSE. Constantly raised amount of MMP-9 released into the tissues may be involved in the degradation of extracellular matrix in the lungs as seen in COPD patients.
MeSH term(s)	Adult ; Aged ; Case-Control Studies ; Cells, Cultured ; Complex Mixtures/toxicity ; Female ; Humans ; L-Lactate Dehydrogenase/metabolism ; Male ; Matrix Metalloproteinase 9/metabolism ; Middle Aged ; Neutrophils/metabolism ; Pulmonary Disease, Chronic Obstructive/blood ; Smoke/adverse effects ; Tobacco Products
Chemical Substances	Complex Mixtures ; Smoke ; L-Lactate Dehydrogenase (EC 1.1.1.27) ; MMP9 protein, human (EC 3.4.24.35) ; Matrix Metalloproteinase 9 (EC 3.4.24.35)
Language	English
Publishing date	2017-12-08
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	6165-7
ISSN	1432-1750 ; 0341-2040
ISSN (online)	1432-1750
ISSN	0341-2040
DOI	10.1007/s00408-017-0077-3
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Article ; Online: Effects of extracellular Hsp70 and cigarette smoke on differentiated THP-1 cells and human monocyte-derived macrophages.

Hulina-Tomašković, Andrea / Somborac-Bačura, Anita / Grdić Rajković, Marija / Bosnar, Martina / Samaržija, Miroslav / Rumora, Lada

Molecular immunology

2019 Volume 111, Page(s) 53–63

Abstract: Extracellular Hsp70 (eHsp70) can act as pro-inflammatory mediator and is elevated in blood of chronic obstructive pulmonary disease (COPD) patients. Most of those patients are smokers, and it was suggested previously that cigarette smoke might induce ... ...

Abstract	Extracellular Hsp70 (eHsp70) can act as pro-inflammatory mediator and is elevated in blood of chronic obstructive pulmonary disease (COPD) patients. Most of those patients are smokers, and it was suggested previously that cigarette smoke might induce Hsp70 secretion from the circulating cells. Therefore, we aimed to explore inflammation-associated effects of cigarette smoke extract (CSE) and its combinations with eHsp70 in monocyte-derived macrophages (MDMs) and THP-1 cell line, used as systemic component models of COPD. We hypothesized that eHsp70 induces inflammation, but that it can also modulate cigarette smoke extract (CSE)-stimulated inflammatory responses. We assessed IL-8 secretion, TLR2, TLR4 and Hsp70 expressions, MAPKs and NF-κB activation, and cytotoxicity after treating the cells with CSE (2.5 and 5%) and its combinations with low-endotoxin recombinant human (rh) Hsp70, used to mimic eHsp70 effects. CSE induced IL-8 secretion from both cell types, but its combinations with rhHsp70 increased IL-8 release compared to CSE alone only from MDMs. In THP-1, combinations of rhHsp70 with 2.5% CSE induced TLR2 and TLR4 mRNA, while 5% CSE decreased TLR2 expression. In MDMs, CSE alone attenuated TLR2, while rhHsp70 increased TLR2 and lowered TLR4 gene expression. Hsp70 mRNA expression was suppressed in THP-1 with rhHsp70 and CSE; however, the same treatments increased its level in MDMs. CSE had cytotoxic effect only on MDMs, but cytotoxicity was reduced in co-treatments with rhHsp70, which also triggered apoptosis. CSE and rhHsp70 activated p38 and JNK, while ERK was activated only by rhHsp70 in MDMs. In THP-1, 2.5% CSE activated ERK, and 5% CSE activated p38. Inhibition of NF-κB and JNK in MDMs, and ERK and JNK in THP-1 cells, attenuated IL-8 release after rhHsp70 treatment. In conclusion, rhHsp70 provoked pro-inflammatory effects and could also modulate inflammatory response to CSE on protein and gene expression levels in THP-1 cells and MDMs, which suggests that eHsp70 might be implicated in systemic inflammation induced by cigarette smoke.
MeSH term(s)	Apoptosis/physiology ; Cell Differentiation/physiology ; Cell Line ; Epithelial Cells/metabolism ; HSP70 Heat-Shock Proteins/metabolism ; Humans ; Inflammation/metabolism ; Interleukin-8/metabolism ; Macrophages/metabolism ; NF-kappa B/metabolism ; Pulmonary Disease, Chronic Obstructive/metabolism ; Signal Transduction/physiology ; Smoke/adverse effects ; Smoking/adverse effects ; Smoking/metabolism ; THP-1 Cells/metabolism ; Nicotiana/adverse effects ; Toll-Like Receptor 4/metabolism
Chemical Substances	HSP70 Heat-Shock Proteins ; Interleukin-8 ; NF-kappa B ; Smoke ; Toll-Like Receptor 4
Language	English
Publishing date	2019-04-11
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	424427-8
ISSN	1872-9142 ; 0161-5890
ISSN (online)	1872-9142
ISSN	0161-5890
DOI	10.1016/j.molimm.2019.04.002
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Article ; Online: Differential expression of heat shock proteins and activation of mitogen-activated protein kinases in A549 alveolar epithelial cells exposed to cigarette smoke extract.

Somborac-Bačura, Anita / Rumora, Lada / Novak, Ruđer / Rašić, Dubravka / Dumić, Jerka / Čepelak, Ivana / Žanić-Grubišić, Tihana

Experimental physiology

2018 Volume 103, Issue 12, Page(s) 1666–1678

Abstract: New findings: What is the central question of this study? What is the effect of cigarette smoke on cell death, oxidative damage, expression of heat shock proteins (HSPs) and activation of mitogen-activated protein kinases (MAPKs) in A549 alveolar ... ...

Abstract	New findings: What is the central question of this study? What is the effect of cigarette smoke on cell death, oxidative damage, expression of heat shock proteins (HSPs) and activation of mitogen-activated protein kinases (MAPKs) in A549 alveolar epithelial cells? What is the main finding and its importance? Cigarette smoke induces cytotoxicity and oxidative damage to A549 cells, increases expression of different HSPs and activates MAPK signalling pathways. This could be related to inflammatory response and apoptosis observed in lungs of patients with smoking-related diseases. Abstract: Cigarette smoking is one of the main risk factors for development of chronic obstructive pulmonary disease (COPD). We previously reported that cigarette smoke (CS) induces damage to proteins and their ineffective degradation. Here, we hypothesize that CS could induce oxidative stress and cytotoxicity in lung epithelial cells through alterations of heat shock protein (HSP) expression and mitogen-activated protein kinase (MAPK) signalling pathways. We exposed A549 alveolar epithelial cells to various concentrations of cigarette smoke extract (CSE). Higher concentrations of CSE caused apoptosis of A549 cells after 4 h, while after 24 h cell viability was decreased, and lactate dehydrogenase in cell culture medium was increased as well as the number of necrotic cells. Concentrations of malondialdehyde (MDA) were elevated, while total thiol groups were decreased. Changes in the expression of HSPs (HSP70, HSP32 and HSP27) were time-dependent. After 6 h, CSE caused an increase in the expression of HSP70 and HSP32, while after 8 h all examined HSPs were up-regulated and remained increased up to 48 h. Treatment of A549 cells with CSE stimulated phosphorylation of extracellular signal-regulated kinase and p38 in a dose-dependent manner, while c-Jun N-terminal kinase activation was not detected. By using specific inhibitors, we demonstrated that MAPKs and HSPs interplay in CSE effects. In conclusion, our results show that MAPKs and HSPs are involved in the mechanism underlying CSE-induced cytotoxicity and oxidative damage to A549 alveolar epithelial cells. These processes could be related to inflammatory response and apoptosis observed in lungs of patients with smoking-related diseases, such as COPD.
MeSH term(s)	A549 Cells ; Alveolar Epithelial Cells/metabolism ; Apoptosis/physiology ; Cell Line, Tumor ; Cell Survival/physiology ; Heat-Shock Proteins/metabolism ; Humans ; L-Lactate Dehydrogenase/metabolism ; Lung/metabolism ; MAP Kinase Signaling System/physiology ; Malondialdehyde/metabolism ; Mitogen-Activated Protein Kinases/metabolism ; Oxidative Stress/physiology ; Pulmonary Disease, Chronic Obstructive/metabolism ; Smoke/adverse effects ; Smoking/metabolism ; Nicotiana/adverse effects ; Up-Regulation/physiology
Chemical Substances	Heat-Shock Proteins ; Smoke ; Malondialdehyde (4Y8F71G49Q) ; L-Lactate Dehydrogenase (EC 1.1.1.27) ; Mitogen-Activated Protein Kinases (EC 2.7.11.24)
Language	English
Publishing date	2018-10-16
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1016295-1
ISSN	1469-445X ; 0958-0670
ISSN (online)	1469-445X
ISSN	0958-0670
DOI	10.1113/EP087038
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