Cathelicidins are peptide components of the innate immune system of mammals. Apart from exerting a direct antibiotic activity, they can also trigger specific defense responses in the host. Their roles in various pathophysiological conditions have been studied, but there is a lack of published information on their expression and activities in the context of mastitis. The aims of this study were to investigate the expression of the bovine cathelicidins BMAP-27, BMAP-28, Bac5, and indolicidin in healthy and infected mammary tissue and in lipopolysaccharide (LPS)-treated cells, to determine their activities against bacteria isolated from bovine mastitis, and to examine their potentials to trigger defense responses in bovine mammary cells. The genes were found to be upregulated in LPS-stimulated neutrophils, but not in infected quarters or epithelial cells. All peptides showed a variably broad spectrum of activity against 28 bacterial isolates from bovine mastitis (MIC values, 0.5 to 32 M), some of which were antibiotic resistant. The activity of each peptide was significantly enhanced when it was pairwise tested with the other peptides, reaching the synergy threshold when indolicidin was present. The bactericidal activity was sensitive to milk components; BMAP-27 and -28 were highly effective in mastitic bovine milk and inhibited in milk from healthy cows. Both peptides were also active in whey and in blood serum and triggered the expression of tumor necrosis factor alpha (TNF-␣) in bovine mammary epithelial cells. Our results indicate multiple roles for the bovine cathelicidins in mastitis, with complementary and mutually enhanced antimicrobial activities against causative pathogens and the capacity to activate host cells.
Designing
small molecules able to break down G4 structures in mRNA
(RG4s) offers an interesting approach to cancer therapy. Here, we
have studied cationic porphyrins (CPs) bearing an alkyl chain up to
12 carbons, as they bind to RG4s while generating reactive oxygen
species upon photoirradiation. Fluorescence-activated cell sorting
(FACS) and confocal microscopy showed that the designed alkyl CPs
strongly penetrate cell membranes, binding to KRAS and NRAS mRNAs under low-abundance cell conditions.
In Panc-1 cells, alkyl CPs at nanomolar concentrations promote a dramatic
downregulation of KRAS and NRAS expression,
but only if photoactivated. Alkyl CPs also reduce the metabolic activity
of pancreatic cancer cells and the growth of a Panc-1 xenograft in
SCID mice. Propidium iodide/annexin assays and caspase 3, caspase
7, and PARP-1 analyses show that these compounds activate apoptosis.
All these data demonstrate that the designed alkyl CPs are efficient
photosensitizers for the photodynamic therapy of ras-driven cancers.
OriginalAntimicrobial and host cell-directed activities of Gly/Ser-rich peptides from salmonid cathelicidins This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
M A N U S C R I P T A C C E P T E D ACCEPTED MANUSCRIPT
M A N U S C R I P T A C C E P T E D ACCEPTED MANUSCRIPT
3Highlights 38• Salmonid cathelicidin-derived peptides show medium-sensitive antimicrobial activity 39• They are not cytotoxic to fish cells and promote proliferation of trout fibroblasts 40• Peptide STF(1-37) potentiates phagocytosis and respiratory burst in trout leukocytes 41• STF(1-37) and β-glucan act synergistically to immunostimulate trout leukocytes 42Cathelicidins, a major family of vertebrate antimicrobial peptides (AMPs), have a recognized role 44 in the first line of defense against infections. They have been identified in several salmonid species, 45 where the putative mature peptides are unusually long and rich in serine and glycine residues, often 46 arranged in short multiple repeats (RLGGGS/RPGGGS) intercalated by hydrophobic motifs. 47Fragments of 24 to 40 residues, spanning specific motifs and conserved sequences in grayling or 48 brown, rainbow and brook trout, were chemically synthesized and examined for antimicrobial 49 activity against relevant Gram-positive and Gram-negative salmonid pathogens, as well as 50 laboratory reference strains. They were not active in complete medium, but showed varying potency 51 and activity spectra in diluted media. Bacterial membrane permeabilization also occurred only 52 under these conditions and was indicated by rapid propidium iodide uptake in peptide-treated 53 bacteria. However, circular dichroism analyses indicated that they did not significantly adopt 54 ordered conformations in membrane-like environments. The peptides were not hemolytic or 55 cytotoxic to trout cells, including freshly purified head kidney leukocytes (HKL) and the 56 fibroblastic RTG-2 cell line. Notably, when exposed to them, HKL showed increased metabolic 57 activity, while a growth-promoting effect was observed on RTG-2 cells, suggesting a functional 58 interaction of salmonid cathelicidins with host cells similar to that shown by mammalian ones. The 59 three most active peptides produced a dose-dependent increase in phagocytic uptake by HKL 60 simultaneously stimulated with bacterial particles. The peptide STF(1-37), selected for further 61 analyses, also enhanced phagocytic uptake in the presence of autologous serum, and increased 62 intracellular killing of live E. coli. Furthermore, when tested on HKL in combination with the 63 immunostimulant β-glucan, it synergistically potentiated both phagocytic up...
This research was conducted to evaluate the potential use of saturated monoglyceride (MG)-based gels in the protection of probiotics upon in vitro digestion.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.