TY  - JOUR
A1  - Shityakov, Sergey
A1  - Salvador, Ellaine
A1  - Pastorin, Giorgia
A1  - Förster, Carola
T1  - Blood-brain barrier transport studies, aggregation, and molecular dynamics simulation of multiwalled carbon nanotube functionalized with fluorescein isothiocyanate
JF  - International Journal of Nanomedicine
N2  - In this study, the ability of a multiwalled carbon nanotube functionalized with fluorescein isothiocyanate (MWCNT-FITC) was assessed as a prospective central nervous system-targeting drug delivery system to permeate the blood-brain barrier. The results indicated that the MWCNT-FITC conjugate is able to penetrate microvascular cerebral endothelial monolayers; its concentrations in the Transwell® system were fully equilibrated after 48 hours. Cell viability test, together with phase-contrast and fluorescence microscopies, did not detect any signs of MWCNT-FITC toxicity on the cerebral endothelial cells. These microscopic techniques also revealed presumably the intracellular localization of fluorescent MWCNT-FITCs apart from their massive nonfluorescent accumulation on the cellular surface due to nanotube lipophilic properties. In addition, the 1,000 ps molecular dynamics simulation in vacuo discovered the phenomenon of carbon nanotube aggregation driven by van der Waals forces via MWCN-TFITC rapid dissociation as an intermediate phase.
KW  - endothelial cells
KW  - cytotoxicity
KW  - blood-brain barrier
KW  - fluorescein isothiocyanate
KW  - aggregation
KW  - molecular dynamics
KW  - fluorescence microscopy
KW  - Transwell® system
KW  - multiwalled carbon nanotube
KW  - mice
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-149233
VL  - 10
ER  - 
TY  - JOUR
A1  - León-Calvijo, María A.
A1  - Leal-Castro, Aura L.
A1  - Almanzar-Reina, Giovanni A.
A1  - Rosas-Pérez, Jaiver E.
A1  - García-Castañeda, Javier E.
A1  - Rivera-Monroy, Zuly J.
T1  - Antibacterial activity of synthetic peptides derived from lactoferricin against Escherichia coli ATCC 25922 and Enterococcus faecalis ATCC 29212
JF  - BioMed Research International
N2  - Peptides derived from human and bovine lactoferricin were designed, synthesized, purified, and characterized using RP-HPLC and MALDI-TOF-MS. Specific changes in the sequences were designed as (i) the incorporation of unnatural amino acids in the sequence, the (ii) reduction or (iii) elongation of the peptide chain length, and (iv) synthesis of molecules with different number of branches containing the same sequence. For each peptide, the antibacterial activity against Escherichia coli ATCC 25922 and Enterococcus faecalis ATCC 29212 was evaluated. Our results showed that Peptides I.2 (RWQWRWQWR) and I.4 ((RRWQWR)\(_{4}\)K\(_{2}\)Ahx\(_{2}\)C\(_{2}\)) exhibit bigger or similar activity against E. coli (MIC 4-33 μM) and E. faecalis (MIC 10-33 μM) when they were compared with lactoferricin protein (LF) and some of its derivate peptides as II.1 (FKCRRWQWRMKKLGA) and IV.1 (FKCRRWQWRMKKLGAPSITCVRRAE). It should be pointed out that Peptides I.2 and I.4, containing the RWQWR motif, are short and easy to synthesize; our results demonstrate that it is possible to design and obtain synthetic peptides that exhibit enhanced antibacterial activity using a methodology that is fast and low-cost and that allows obtaining products with a high degree of purity and high yield.
KW  - bovine lactoferricin
KW  - antimicrobial activity
KW  - infection
KW  - spectrum
KW  - mice
KW  - cells
KW  - inhibit
KW  - derivatives
KW  - loop region
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144591
IS  - 453826
ER  - 
TY  - JOUR
A1  - Sagiv, Jitka Y.
A1  - Michaeli, Janna
A1  - Assi, Simaan
A1  - Mishalian, Inbal
A1  - Kisos, Hen
A1  - Levy, Liran
A1  - Damti, Pazzit
A1  - Lumbroso, Delphine
A1  - Polyansky, Lola
A1  - Sionov, Ronit V.
A1  - Ariel, Amiram
A1  - Hovav, Avi-Hai
A1  - Henke, Erik
A1  - Fridlender, Zvi G.
A1  - Granot, Zvi
T1  - Phenotypic diversity and plasticity in circulating neutrophil subpopulations in cancer
JF  - Cell Reports
N2  - Controversy surrounds neutrophil function in cancer because neutrophils were shown to provide both pro-and antitumor functions. We identified a heterogeneous subset of low-density neutrophils (LDNs) that appear transiently in self-resolving inflammation but accumulate continuously with cancer progression. LDNs display impaired neutrophil function and immunosuppressive properties, characteristics that are in stark contrast to those of mature, high-density neutrophils (HDNs). LDNs consist of both immature myeloid-derived suppressor cells (MDSCs) and mature cells that are derived from HDNs in a TGF-beta-dependent mechanism. Our findings identify three distinct populations of circulating neutrophils and challenge the concept that mature neutrophils have limited plasticity. Furthermore, our findings provide a mechanistic explanation to mitigate the controversy surrounding neutrophil function in cancer.
KW  - tumorigenic properties
KW  - innate immunity
KW  - adenocarcinoma
KW  - inhibition
KW  - model
KW  - suppressor cells
KW  - tumor
KW  - TGF-beta
KW  - mice
KW  - lung
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144102
VL  - 10
IS  - 4
ER  - 
TY  - JOUR
A1  - Konrad, Franziska M.
A1  - Bury, Annette
A1  - Schick, Martin A.
A1  - Ngamsri, Kristian-Christos
A1  - Reutershan, Jörg
T1  - The Unrecognized Effects of Phosphodiesterase 4 on Epithelial Cells in Pulmonary Inflammation
JF  - PLoS ONE
N2  - Acute pulmonary inflammation is characterized by migration of polymorphonuclear neutrophils (PMNs) into the different compartments of the lung, passing an endothelial and epithelial barrier. Recent studies showed evidence that phosphodiesterase (PDE) 4-inhibitors stabilized endothelial cells. PDE4B and PDE4D subtypes play a pivotal role in inflammation, whereas blocking PDE4D is suspected to cause gastrointestinal side effects. We thought to investigate the particular role of the PDE4-inhibitors roflumilast and rolipram on lung epithelium. Acute pulmonary inflammation was induced by inhalation of LPS. PDE4-inhibitors were administered i.p. or nebulized after inflammation. The impact of PDE4-inhibitors on PMN migration was evaluated in vivo and in vitro. Microvascular permeability, cytokine levels, and PDE4B and PDE4D expression were analyzed. In vivo, both PDE4-inhibitors decreased transendothelial and transepithelial migration even when administered after inflammation, whereas roflumilast showed a superior effect compared to rolipram on the epithelium. Both inhibitors decreased TNF\(\alpha\), IL6, and CXCL2/3. CXCL1, the strong PMN chemoattractant secreted by the epithelium, was significantly more reduced by roflumilast. In vitro assays with human epithelium also emphasized the pivotal role of roflumilast on the epithelium. Additionally, LPS-induced stress fibers, an essential requirement for a direct migration of PMNs into the alveolar space, were predominantly reduced by roflumilast. Expression of PDE4B and PDE4D were both increased in the lungs by LPS, PDE4-inhibitors decreased mainly PDE4B. The topical administration of PDE4-inhibitors was also effective in curbing down PMN migration, further highlighting the clinical potential of these compounds. In pulmonary epithelial cells, both subtypes were found coexistent around the nucleus and the cytoplasm. In these epithelial cells, LPS increased PDE4B and, to a lesser extend, PDE4D, whereas the effect of the inhibitors was prominent on the PDE4B subtype. In conclusion, we determined the pivotal role of the PDE4-inhibitor roflumilast on lung epithelium and emphasized its main effect on PDE4B in hyperinflammation.
KW  - acute lung injury
KW  - PDE4-inhibitor roflumilast
KW  - GRO alpha
KW  - expression
KW  - 4D
KW  - respiratory distress syndrome
KW  - mice
KW  - infiltration
KW  - rolipram
KW  - disease
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-143203
VL  - 10
IS  - 4
ER  - 
TY  - JOUR
A1  - Kleikers, Pamela W. M.
A1  - Hooijmans, Carlijn
A1  - Göb, Eva
A1  - Langhauser, Friederike
A1  - Rewell, Sarah S. J.
A1  - Radermacher, Kim
A1  - Ritskes-Hoitinga, Merel
A1  - Howells, David W.
A1  - Kleinschnitz, Christoph
A1  - Schmidt, Harald H. H. W.
T1  - A combined pre-clinical meta-analysis and randomized confirmatory trial approach to improve data validity for therapeutic target validation
JF  - Scientific Reports
N2  - Biomedical research suffers from a dramatically poor translational success. For example, in ischemic stroke, a condition with a high medical need, over a thousand experimental drug targets were unsuccessful. Here, we adopt methods from clinical research for a late-stage pre-clinical meta-analysis (MA) and randomized confirmatory trial (pRCT) approach. A profound body of literature suggests NOX\(_{2}\) to be a major therapeutic target in stroke. Systematic review and MA of all available NOX\(_{2}\)\(^{-/y}\) studies revealed a positive publication bias and lack of statistical power to detect a relevant reduction in infarct size. A fully powered multi-center pRCT rejects NOX\(_{2}\) as a target to improve neurofunctional outcomes or achieve a translationally relevant infarct size reduction. Thus stringent statistical thresholds, reporting negative data and a MA-pRCT approach can ensure biomedical data validity and overcome risks of bias.
KW  - focal cerebral ischemia
KW  - darbepoetin alpha
KW  - mice
KW  - translational stroke research
KW  - colony-stimulating factor
KW  - NADPH oxidase inhibitors
KW  - chronic kidney disease
KW  - diabetes mellitus
KW  - oxidative stress
KW  - search filter
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-151401
VL  - 5
IS  - 13428
ER  - 
TY  - JOUR
A1  - Singh, Amit K.
A1  - Kingston, Joseph J.
A1  - Gupta, Shishir K.
A1  - Batra, Harsh V.
T1  - Recombinant Bivalent Fusion Protein rVE Induces CD4+ and CD8+ T-Cell Mediated Memory Immune Response for Protection Against Yersinia enterocolitica Infection
JF  - Frontiers in Microbiology
N2  - Studies investigating the correlates of immune protection against Yersinia infection have established that both humoral and cell mediated immune responses are required for the comprehensive protection. In our previous study, we established that the bivalent fusion protein (rVE) comprising immunologically active regions of Y pestis LcrV (100-270 aa) and YopE (50-213 aa) proteins conferred complete passive and active protection against lethal Y enterocolitica 8081 challenge. In the present study, cohort of BALB/c mice immunized with rVE or its component proteins rV, rE were assessed for cell mediated immune responses and memory immune protection against Y enterocolitica 8081 rVE immunization resulted in extensive proliferation of both CD4 and CD8 T cell subsets; significantly high antibody titer with balanced IgG1: IgG2a/IgG2b isotypes (1:1 ratio) and up regulation of both Th1 (INF-\(\alpha\), IFN-\(\gamma\), IL 2, and IL 12) and Th2 (IL 4) cytokines. On the other hand, rV immunization resulted in Th2 biased IgG response (11:1 ratio) and proliferation of CD4+ T-cell; rE group of mice exhibited considerably lower serum antibody titer with predominant Th1 response (1:3 ratio) and CD8+ T-cell proliferation. Comprehensive protection with superior survival (100%) was observed among rVE immunized mice when compared to the significantly lower survival rates among rE (37.5%) and rV (25%) groups when IP challenged with Y enterocolitica 8081 after 120 days of immunization. Findings in this and our earlier studies define the bivalent fusion protein rVE as a potent candidate vaccine molecule with the capability to concurrently stimulate humoral and cell mediated immune responses and a proof of concept for developing efficient subunit vaccines against Gram negative facultative intracellular bacterial pathogens.
KW  - I-tasser
KW  - Yersinia enterocolitica
KW  - memory immune responses
KW  - cytokine profiling
KW  - CD8+T cells
KW  - CD4+T cells
KW  - recombinant protein rVE
KW  - resistance
KW  - pneumonic plague
KW  - pestis infection
KW  - nonhuman-primates
KW  - III secretion
KW  - V-antigen
KW  - mice
KW  - vaccine
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-136114
VL  - 6
IS  - 1407
ER  -