34 research outputs found
Effect of steroids on transcription and secretion of Gal-1 by the human trophoblast cell line in vitro
Galectin-1 (Gal-1) is a lectin with recently documented pro-invasive function in trophoblasts in vitro, whose regulation is currently insufficiently known. The potential involvement of steroid hormones, synthetic glucocorticoid dexamethasone (DEX), the sex steroid progesterone (PRG) and mifepristone (RU486) in the regulation of Gal-1 in the trophoblast-derived cell line HTR-8/SVneo was investigated. Gal-1 mRNA levels were assessed by real-time PCR. The effect on secretion of Gal-1 into the culture media was followed using the SELDI-TOF protein chip array. We present evidence that DEX and RU486 significantly reduced Gal-1 in the HTR-8/SVneo cell line at the mRNA level. In addition, trophoblast-derived HTR-8/SVneo cells were shown to secrete detectable Gal-1 protein, which was only slightly increased by PRG. The potential clinical relevance of these findings remains to be determined
Effect of steroids on transcription and secretion of Gal-1 by the human trophoblast cell line in vitro
Galectin-1 (Gal-1) is a lectin with recently documented pro-invasive function
in trophoblasts in vitro, whose regulation is currently insufficiently known.
The potential involvement of steroid hormones, synthetic glucocorticoid
dexamethasone (DEX), the sex steroid progesterone (PRG) and mifepristone
(RU486) in the regulation of Gal-1 in the trophoblast-derived cell line
HTR-8/SVneo was investigated. Gal-1 mRNA levels were assessed by real-time
PCR. The effect on secretion of Gal-1 into the culture media was followed
using the SELDI-TOF protein chip array. We present evidence that DEX and
RU486 significantly reduced Gal-1 in the HTR-8/SVneo cell line at the mRNA
level. In addition, trophoblast-derived HTR-8/SVneo cells were shown to
secrete detectable Gal-1 protein, which was only slightly increased by PRG.
The potential clinical relevance of these findings remains to be determined.
[Projekat Ministarstva nauke Republike Srbije, br. 173004
Može li salivarni karcioembrionalni antigen biti koristan za dijagnozu Sjƶgrenovog sindroma?
Didehydroroflamycoin pentaene macrolide family from Streptomyces durmitorensis MS405(T): production optimization and antimicrobial activity
AimsThe aim of this study was to improve production of pentaene 32,33-didehydroroflamycoin (DDHR) in Streptomyces durmitorensis MS405 strain to obtain quantities sufficient for in depth analysis of antimicrobial properties. Methods and ResultsThrough classical medium optimization conditions for stable growth, DDHR production within 7days of incubation was established. Yields of 215mgl(-1) were achieved in shake flask experiments in complex medium with mannitol as the primary carbon source. DDHR had poor antibacterial activity with minimal inhibitory concentrations (MIC) of 400gml(-1) for Staphylococcus aureus and Bacillus subtilis, while MIC of 70gml(-1) was determined for Candida albicans. Using flow cytometry and fluorescent microscopy, it was demonstrated that DDHR induced membrane damage in C.albicans followed by cell death. Combination studies with known antifungal nystatin showed that DDHR is a promising agent for the development of novel antimycotic treatments potentially less toxic for human cells. ConclusionsPentaene didehydroroflamycoin has no antibacterial activity but can be further developed for the application in antifungal therapy. Significance and Impact of the StudyThis study is the first report on the stable and production in high yields of a novel pentaene family that acts on Candida cell membranes and can be used in combination with known antifungals. Polyenes are still antifungal antibiotics of choice, and therefore, isolation and production of new lead structures are highly significant
Supplementary data for article: StankoviÄ, N.; Å eneroviÄ, L.; Bojic-Trbojevic, Z.; Vuckovic, I.; Vicovac, L.; Vasiljevic, B.; NikodinoviÄ-RuniÄ, J. Didehydroroflamycoin Pentaene Macrolide Family from Streptomyces Durmitorensis MS405(T): Production Optimization and Antimicrobial Activity. Journal of Applied Microbiology 2013, 115 (6), 1297ā1306. https://doi.org/10.1111/jam.12326
Supporting information for: [https://doi.org/10.1111/jam.12326]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1438]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/3486
Supplementary data for article: StankoviÄ, N.; Å eneroviÄ, L.; Bojic-Trbojevic, Z.; Vuckovic, I.; Vicovac, L.; Vasiljevic, B.; NikodinoviÄ-RuniÄ, J. Didehydroroflamycoin Pentaene Macrolide Family from Streptomyces Durmitorensis MS405(T): Production Optimization and Antimicrobial Activity. Journal of Applied Microbiology 2013, 115 (6), 1297ā1306. https://doi.org/10.1111/jam.12326
Supporting information for: [https://doi.org/10.1111/jam.12326]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1438]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/3486
Reticulocyte response in paired blood samples of Babesia canis infected young and adult dogs
Acute B. canis infection can lead to an acute phase reaction (APR) in dogs. The parasite invades red blood cells causing
anemia through immune-mediated hemolysis and possible erythropoietic suppression. A regenerative response of the
erythroid lineage during the babesiosis has not been described in extension. This research examines hematologic parameters
focusing on the absolute reticulocyte count and apolipoprotein A I (ApoA I) level on the day of admission and 14 days after
treatment with imidocarb-dipropionate in young (n=11) and adult (n=11) dogs naturally infected with B. canis. Metabolic
and inflammatory processes were characterized by analyzing protein and lipid profiles, as well as ApoA I at specified time
points. Automated analyzers were used to determine complete blood count and biochemical parameters, while ApoA I was
assessed using radioimmunoassay. The reticulocyte count was determined using a manual method by means of supravital
staining. Both young and adult dogs with acute B. canis infection showed non-regenerative anemia without difference.
Fourteen days after successful treatment with imidocarb-dipropionate, the anemia was corrected and a high reticulocyte
count was observed (p<0.05). This indicates that the erythroid regenerative response was efficient in young and adult dogs,
although vital signs, leukocyte count and triglyceride concentration suggest a more intense APR in young dogs. A decrease
in ApoA I in both groups 14 days after treatment (p<0.01) confirmed that this lipoprotein acts as a positive acute-phase
protein in acute B. canis infection in dogs, but further studies are needed to connect its role in erythroid lineage regeneration
Oleuropein Attenuates Oxidative Stress in Human Trophoblast Cells
Olive-derived bioactive compound oleuropein was evaluated against damage induced by hydrogen peroxide in human trophoblast cells in vitro, by examining the changes in several markers implicated in oxidative stress interactions in the placenta. Trophoblast HTR-8/SVneo cells were preincubated with OLE at 10 and 100 ĀµM and exposed to H2O2, as a model of oxidative stress. Protein and lipid peroxidation, as well as antioxidant enzymesā activity, were determined spectrophotometrically, and DNA damage was evaluated by comet assay. iNOS protein expression was assessed by Western blot, while the mRNA expression of pro- and anti-apoptotic genes BAX and BCL2 and transcription factor NFE2L2, as well as cytokines IL-6 and TNF Ī± were determined by qPCR. Oleuropein demonstrated cytoprotective effects against H2O2 in trophoblast cells by significantly improving the antioxidant status and preventing protein and lipid damage, as well as reducing the iNOS levels. OLE reduced the mRNA expression of IL-6 and TNF Ī±, however, it did not influence the expression of NFE2L2 or the BAX/BCL2 ratio after H2O2 exposure. Oleuropein per se did not lead to any adverse effects in HTR-8/SVneo cells under the described conditions, confirming its safety in vitro. In conclusion, it significantly attenuated oxidative damage and restored antioxidant functioning, confirming its protective role in trophoblast. Ā© 2023 by the authors
Association of acute Babesia canis infection and serum lipid, lipoprotein, and apoprotein concentrations in dogs
Background: Babesia canis infection induces a marked acute phase response (APR) that might be associated with alteration in lipid and lipoprotein metabolism and disease prognosis. Hypothesis: Dogs with B. canis-induced APR develop dyslipidemia with altered lipoprotein concentration and morphology. Animals: Twenty-nine client-owned dogs with acute B. canis infection and 10 clinically healthy control dogs. Methods: Observational cross-sectional study. Serum amyloid A (SAA) was measured using ELISA. Cholesterol, phospholipids, and triglycerides were determined biochemically. Lipoproteins were separated using agarose gel electrophoresis. Lipoprotein diameter was assessed by polyacrylamide gradient gel electrophoresis; correlation with ApoA-1 (radioimmunoassay) and SAA was determined. Results: Dogs with B. canis infection had a marked APR (median SAA, 168.3 Ī¼g/mL; range, 98.1-716.2 Ī¼g/mL) compared with controls (3.2 Ī¼g/mL, 2.0-4.2 Ī¼g/mL) (P lt .001). Dogs with B. canis infection had significantly lower median cholesterol (4.79 mmol/L, 1.89-7.64 mmol/L versus 6.15 mmol/L, 4.2-7.4 mmol/L) (P =.02), phospholipid (4.64 mmol/L, 2.6-6.6 mmol/L versus 5.72 mmol/L, 4.68-7.0 mmol/L) (P =.02), and Ī±-lipoproteins (77.5%, 27.7%-93.5% versus 89.2%, 75.1%-93.5%) (P =.04), and higher ApoA-1 (1.36 U, 0.8-2.56 U versus 0.95 U, 0.73-1.54 U) concentrations (P =.02). Serum amyloid A correlated with high-density lipoproteins (HDLs) diameter (rho =.43; P =.03) and ApoA-1 (rho =.63, P lt .001). Conclusions and Clinical Importance: Major changes associated with B. canis-induced APR in dogs are related to concentration, composition, and morphology of HDL particles pointing to an altered reverse cholesterol transport. Parallel ApoA-1 and SAA concentration increase is a unique still unexplained pathophysiological finding
Galectin-1 Is Part of Human Trophoblast Invasion Machinery - A Functional Study In Vitro
Interactions of glycoconjugates with endogenous galectins, have been long proposed to participate in several reproductive processes including implantation. In human placenta gal-1, gal-3, gal-8, and gal-13 proteins are known to be present. Each of them has been proposed to play multiple functions, but so far no clear picture has emerged. We hypothesized that gal-1 participates in trophoblast invasion, and conducted Matrigel invasion assay using isolated cytotrophoblast from first trimester placenta and HTR-8/SVneo cell line to test it.<0.001) by Ox-gal-1 at 1 Āµg/ml. Both sets of results confirmed involvement of gal-1 in trophoblast invasion. Galectin profile of isolated cytotrophoblast and HTR-8/SVneo cells was established using RT-PCR and real-time PCR and found to consist of gal-1, gal-3 and gal-8 for both cell types. Only gal-1 was located at the trophoblast cell membrane, as determined by FACS analysis, which is consistent with the results of the functional tests.These findings qualify gal-1 as a member of human trophoblast cell invasion machinery