| 1. |
- Hernroth, Bodil, 1951-, et al.
(författare)
-
Impact of ocean acidification on antimicrobial activity in gills of the blue mussel (Mytilus edulis)
- 2016
-
Ingår i: Fish and Shellfish Immunology. - : Elsevier. - 1050-4648 .- 1095-9947. ; 55, s. 452-459
-
Tidskriftsartikel (refereegranskat)abstract
- Here, we aimed to investigate potential effects of ocean acidification on antimicrobial peptide (AMP) activity in the gills of Mytilus edulis, as gills are directly facing seawater and the changing pH (predicted to be reduced from ∼8.1 to ∼7.7 by 2100). The AMP activity of gill and haemocyte extracts was compared at pH 6.0, 7.7 and 8.1, with a radial diffusion assay against Escherichia coli. The activity of the gill extracts was not affected by pH, while it was significantly reduced with increasing pH in the haemocyte extracts. Gill extracts were also tested against different species of Vibrio (V. parahaemolyticus, V. tubiashii, V. splendidus, V. alginolyticus) at pH 7.7 and 8.1. The metabolic activity of the bacteria decreased by ∼65–90%, depending on species of bacteria, but was, as in the radial diffusion assay, not affected by pH. The results indicated that AMPs from gills are efficient in a broad pH-range. However, when mussels were pre-exposed for pH 7.7 for four month the gill extracts presented significantly lower inhibit of bacterial growth. A full in-depth proteome investigation of gill extracts, using LC-Orbitrap MS/MS technique, showed that among previously described AMPs from haemocytes of Mytilus, myticin A was found up-regulated in response to lipopolysaccharide, 3 h post injection. Sporadic occurrence of other immune related peptides/proteins also pointed to a rapid response (0.5–3 h p.i.). Altogether, our results indicate that the gills of blue mussels constitute an important first line defence adapted to act at the pH of seawater. The antimicrobial activity of the gills is however modulated when mussels are under the pressure of ocean acidification, which may give future advantages for invading pathogens.
|
|
| 2. |
- Bauden, Monika, et al.
(författare)
-
In vitro cytotoxicity evaluation of HDAC inhibitor Apicidin in pancreatic carcinoma cells subsequent time and dose dependent treatment
- 2015
-
Ingår i: Toxicology Letters. - : Elsevier. - 0378-4274 .- 1879-3169. ; 236:1, s. 8-15
-
Tidskriftsartikel (refereegranskat)abstract
- Apicidin is a potent histone deacetylase inhibitor (HDACI) that selectively binds to histone deacetylases (HDACs) class I and interferes with the deacetylation process, which results in modification of acetylation level of cellular proteins. The aim of the study was to investigate the potential time and dose dependent cytotoxicity of the test compound, Apicidin, in pancreatic cancer cells Capan-1 and Panc-1 as well as estimate maximal tolerable dose (MTD) of the test agent and determine EC50 using four complementary colorimetric cytotoxicity or viability assays. The cells were treated with increasing concentrations of Apicidin (0-5000nM) for 2, 4 and 6h (short term exposure) or 24, 48 and 72h (long term exposure) before conducting cytotoxic analyses with lactate dehydrogenase assay or viability analyses with sulforhodamine B (SRB), methyl tetrazolium (MTT) and crystal violet (CV) assays. In order to investigate whether Apicidin irreversibly affects the cells already during the short term exposure, the medium containing Apicidin was removed and replaced with fresh culturing medium after 6h of treatment. The cells were then incubated for additional 24, 48 or 72h before carrying out the analysis. The results obtained from cytotoxicity and viability assays indicated, that Apicidin was well tolerated by both cell lines at concentrations below 100nM at any given time point and at all applied concentrations during the short term (6h or less) treatment. Continuous prolonged term exposures (48h or greater) of the cells to Apicidin with concentration exceeding 100nM resulted in significantly increasing cytotoxicity and sustained significant loss of cell viability. Moreover, long term exposure of pancreatic cancer cells Capan-1 and Panc-1 to Apicidin concentrations exceeding 100nM showed an initial anti-proliferative effect before cytotoxicity onset. In summary, MTD was exposure time dependent and estimated to 100nM for long term treatment and to at least 5000nM for treatment not greater than 6h. EC50 concentration of Apicidin was established after long term treatment, however with some variation when comparing the different assays and cell lines. Results from this study may encourage reinvestigating the capacity of potent HDACI Apicidin as an attractive agent for interfering with the deacetylation process catalyzed by HDACs for potential pancreatic cancer intervention.
|
|
| 3. |
- Bauden, Monika, et al.
(författare)
-
In vitro cytotoxicity evaluation of HDAC inhibitor Apicidin in pancreatic carcinoma cells subsequent time and dose dependent treatment
- 2015
-
Ingår i: Toxicology Letters. - : Elsevier BV. - 0378-4274 .- 1879-3169. ; 236:1, s. 8-15
-
Tidskriftsartikel (refereegranskat)abstract
- Apicidin is a potent histone deacetylase inhibitor (HDACI) that selectively binds to histone deacetylases (HDACs) class I and interferes with the deacetylation process, which results in modification of acetylation level of cellular proteins. The aim of the study was to investigate the potential time and dose dependent cytotoxicity of the test compound, Apicidin, in pancreatic cancer cells Capan-1 and Panc-1 as well as estimate maximal tolerable dose (MTD) of the test agent and determine EC50 using four complementary colorimetric cytotoxicity or viability assays. The cells were treated with increasing concentrations of Apicidin (0-5000nM) for 2, 4 and 6h (short term exposure) or 24, 48 and 72h (long term exposure) before conducting cytotoxic analyses with lactate dehydrogenase assay or viability analyses with sulforhodamine B (SRB), methyl tetrazolium (MTT) and crystal violet (CV) assays. In order to investigate whether Apicidin irreversibly affects the cells already during the short term exposure, the medium containing Apicidin was removed and replaced with fresh culturing medium after 6h of treatment. The cells were then incubated for additional 24, 48 or 72h before carrying out the analysis. The results obtained from cytotoxicity and viability assays indicated, that Apicidin was well tolerated by both cell lines at concentrations below 100nM at any given time point and at all applied concentrations during the short term (6h or less) treatment. Continuous prolonged term exposures (48h or greater) of the cells to Apicidin with concentration exceeding 100nM resulted in significantly increasing cytotoxicity and sustained significant loss of cell viability. Moreover, long term exposure of pancreatic cancer cells Capan-1 and Panc-1 to Apicidin concentrations exceeding 100nM showed an initial anti-proliferative effect before cytotoxicity onset. In summary, MTD was exposure time dependent and estimated to 100nM for long term treatment and to at least 5000nM for treatment not greater than 6h. EC50 concentration of Apicidin was established after long term treatment, however with some variation when comparing the different assays and cell lines. Results from this study may encourage reinvestigating the capacity of potent HDACI Apicidin as an attractive agent for interfering with the deacetylation process catalyzed by HDACs for potential pancreatic cancer intervention.
|
|
| 4. |
- Cabaleiro-Lago, Celia, et al.
(författare)
-
Recent Advances in Molecularly Imprinted Polymers and Their Disease-Related Applications
- 2023
-
Ingår i: Polymers. - : MDPI. - 2073-4360. ; 15:21, s. 4199-4199
-
Forskningsöversikt (refereegranskat)abstract
- Molecularly imprinted polymers (MIPs) and the imprinting technique provide polymeric material with recognition elements similar to natural antibodies. The template of choice (i.e., the antigen) can be almost any type of smaller or larger molecule, protein, or even tissue. There are various formats of MIPs developed for different medical purposes, such as targeting, imaging, assay diagnostics, and biomarker detection. Biologically applied MIPs are widely used and currently developed for medical applications, and targeting the antigen with MIPs can also help in personalized medicine. The synthetic recognition sites of the MIPs can be tailor-made to function as analytics, diagnostics, and drug delivery systems. This review will cover the promising clinical applications of different MIP systems recently developed for disease diagnosis and treatment.
|
|
| 5. |
- Czernekova, Michaela, et al.
(författare)
-
Primary Culture of Tardigrade Storage Cells from Richtersius coronifer Richters, 1903
- 2016
-
Konferensbidrag (refereegranskat)abstract
- Coelomocytes are macrophage-like cells in the body cavity or the coelomic spaces of many invertebrates and play major roles in their physiology and immunology. Their structure, function and diversity, however, is still poorly understood.Tardigrades are micrometazoans inhabiting a wide variety of environments and with an ability to survive extreme conditions. Coelomocytes (“storage cells”) represent an important part of tardigrade physiology, storing and distributing energy and possibly also having immunological functions. Few studies of tardigrade cell biology have been reported and neither primary nor continuous cell cultures have been established. Tardigrades are normally found and also cultured in an environment rich in microorganisms, some of which may even be of symbiotic value.In this study we have tried to establish a primary culture of storage cells in the eutardigrade Richtersius coronifer. Different cell media and concentrations of fetal bovine serum (FBS) were tested. Extracting cells from the tardigrades in an antiseptical environment is challenging since it has to be done under a microscope and contamination from the tardigrades surface is also a problem. To avoid this we tried culturing with high concentrations of antibiotics and antimycotics. We managed to keep the cells viable for up to 18 days in Grace insect medium with 10 % FBS at 20-22°C. The medium was changed every third day. 10x Antibiotic-Antimycotic and 5x of Penicillin-Streptomycin were used to minimize contamination. These concentrations reduce the bacterial abundance, but contamination with fungi was still an issue. Cell morphology evaluation was performed daily and no obvious toxic effects on the cells was observed. Cell viability and cell division were evaluated with Trypan blue staining and cell counting in a haemocytometer. The results indicate that the cells are viable and that some cell division occurs, however more studies need to be performed to confirm this. Still, this study provides the first evidence that primary cultures of storage cells from tardigrades are possible to establish, but the culturing method has to be refined to avoid contamination.
|
|
| 6. |
- Czernekova, Michaela, et al.
(författare)
-
Primary Culture of Tardigrade Storage Cells from Richtersius coronifer Richters, 1903
- 2016
-
Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Coelomocytes are macrophage-like cells in the body cavity or the coelomic spaces of many invertebrates and play major roles in their physiology and immunology. Their structure, function and diversity, however, is still poorly understood. Tardigrades are micrometazoans inhabiting a wide variety of environments and with an ability to survive extreme conditions. Coelomocytes (“storage cells”) represent an important part of tardigrade physiology, storing and distributing energy and possibly also having immunological functions. Few studies of tardigrade cell biology have been reported and neither primary nor continuous cell cultures have been established. Tardigrades are normally found and also cultured in an environment rich in microorganisms, some of which may even be of symbiotic value. In this study we have tried to establish a primary culture of storage cells in the eutardigrade Richtersius coronifer. Different cell media and concentrations of fetal bovine serum (FBS) were tested. Extracting cells from the tardigrades in an antiseptical environment is challenging since it has to be done under a microscope and contamination from the tardigrades surface is also a problem. To avoid this we tried culturing with high concentrations of antibiotics and antimycotics. We managed to keep the cells viable for up to 18 days in Grace insect medium with 10 % FBS at 20-22°C. The medium was changed every third day. 10x Antibiotic-Antimycotic and 5x of Penicillin-Streptomycin were used to minimize contamination. These concentrations reduce the bacterial abundance, but contamination with fungi was still an issue. Cell morphology evaluation was performed daily and no obvious toxic effects on the cells was observed. Cell viability and cell division were evaluated with Trypan blue staining and cell counting in a haemocytometer. The results indicate that the cells are viable and that some cell division occurs, however more studies need to be performed to confirm this. Still, this study provides the first evidence that primary cultures of storage cells from tardigrades are possible to establish, but the culturing method has to be refined to avoid contamination.
|
|
| 7. |
- El-Schich, Zahra, et al.
(författare)
-
Induction of morphological changes in death-induced cancer cells monitored by holographic microscopy
- 2015
-
Ingår i: Journal of Structural Biology. - : Elsevier. - 1047-8477 .- 1095-8657. ; 189:3, s. 207-212
-
Tidskriftsartikel (refereegranskat)abstract
- We are using the label-free technique of holographic microscopy to analyze cellular parameters including cell number, confluence, cellular volume and area directly in the cell culture environment. We show that death-induced cells can be distinguished from untreated counterparts by the use of holographic microscopy, and we demonstrate its capability for cell death assessment. Morphological analysis of two representative cell lines (L929 and DU145) was performed in the culture flasks without any prior cell detachment. The two cell lines were treated with the anti-tumour agent etoposide for one to three days. Measurements by holographic microscopy showed significant differences in average cell number, confluence, volume and area when comparing etoposide-treated with untreated cells. The cell volume of the treated cell lines was initially increased at early time-points. By time, cells decreased in volume, especially when treated with high doses of etoposide. In conclusion, we have shown that holographic microscopy allows label-free and completely non-invasive morphological measurements of cell growth, viability and death. Future applications could include real-time monitoring of these holographic microscopy parameters in cells in response to clinically relevant compounds.
|
|
| 8. |
|
|
| 9. |
|
|
| 10. |
- Hadzic, Radinka, et al.
(författare)
-
alpha1-Antitrypsin inhibits Moraxella catarrhalis MID protein-induced tonsillar B cell proliferation and IL-6 release.
- 2006
-
Ingår i: Immunology Letters. - : Elsevier BV. - 0165-2478 .- 1879-0542. ; 102:2, s. 141-147
-
Tidskriftsartikel (refereegranskat)abstract
- alpha 1-Antitrypsin (AAT) is a major circulating and tissues inhibitor of serine proteinases implicated in the regulation of inflammation and host defence. There is now increasing evidence that AAT may also exhibit anti-inflammatory activities independent of its protease inhibitor function. This study was undertaken to investigate the effects of native (inhibitory) and polymerized (non-inhibitory) forms of AAT on MID (Moraxella IgD binding protein)-induced human tonsillar B cell activation in vitro. We found that 0.5 mu g/ml MID induces B cell proliferation and stimulates IL-6 release (p < 0.001) relative to non-stimulated controls. Both native and polymerized AAT (0.5 mg/ml) inhibited MID-stimulated B cell proliferation in a similar manner (by 70%, p < 0.001), whereas MID-induced IL-6 release was more strongly suppressed by polymefized (9.9-fold, p < 0.001) as compared to native AAT (2.8-fold, p < 0.01). Electrophoretic analysis of cell culture media did not indicate any interaction between AAT and MID, and flow cytometry data showed no competition for the same receptor. The effects of AATs were observed whether added together with MID or 2 h after MID-addition to cell cultures. Thus, our data demonstrate that AAT inhibits MID-induced B cell activation in vitro that is unrelated to its protease inhibitory activity and is not dependent on MID binding to the cell surface.
|
|
