The use of human serum in supporting the in vitro and in vivo proliferation of human conjunctival epithelial cells

The growing problem of antimicrobial resistance highlights the need for alternative strategies to combat infections. From this perspective, there is a considerable interest in natural molecules obtained from different sources, which are shown to be active against microorganisms, either alone or in association with conventional drugs. In this paper, peptides with the same sequence of fragments, found in human serum, derived from physiological proteins, were evaluated for their antifungal activity. A 13-residue peptide, representing the 597–609 fragment within the albumin C-terminus, was proved to exert a fungicidal activity in vitro against pathogenic yeasts and a therapeutic effect in vivo in the experimental model of candidal infection in Galleria mellonella. Studies by confocal microscopy and transmission and scanning electron microscopy demonstrated that the peptide penetrates and accumulates in Candida albicans cells, causing gross morphological alterations in cellular structure. These findings add albumin to the group of proteins, which already includes hemoglobin and antibodies, that could give rise to cryptic antimicrobial fragments, and could suggest their role in anti-infective homeostasis. The study of bioactive fragments from serum proteins could open interesting perspectives for the development of new antimicrobial molecules derived by natural sources.

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Three-dimensional Growth of Renal Epithelial Cells in Vitro: A Tool in Toxicity Testing

Alternatives to Laboratory Animals ◽

10.1177/026119299302100212 ◽

1993 ◽

Vol 21 (2) ◽

pp. 191-195 ◽

Cited By ~ 1

Author(s):

Knut-Jan Andersen ◽

Erik Ilsø Christensen ◽

Hogne Vik

Keyword(s):

Epithelial Cells ◽

Three Dimensional ◽

Toxicity Testing ◽

Renal Tissue ◽

Epithelial Cell Line ◽

Multicellular Spheroids ◽

Renal Epithelial Cell ◽

Renal Epithelial Cells

The tissue culture of multicellular spheroids from the renal epithelial cell line LLC-PK1 (proximal tubule) is described. This represents a biological system of intermediate complexity between renal tissue in vivo and simple monolayer cultures. The multicellular structures, which show many similarities to kidney tubules in vivo, including a vectorial water transport, should prove useful for studying the potential nephrotoxicity of drugs and chemicals in vitro. In addition, the propagation of renal epithelial cells as multicellular spheroids in serum-free culture may provide information on the release of specific biological parameters, which may be suppressed or masked in serum-supplemented media.

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68Ga, 44Sc and 177Lu-labeled AAZTA5-PSMA-617: synthesis, radiolabeling, stability and cell binding compared to DOTA-PSMA-617 analogues

EJNMMI Radiopharmacy and Chemistry ◽

10.1186/s41181-020-00107-8 ◽

2020 ◽

Vol 5 (1) ◽

Author(s):

Jean-Philippe Sinnes ◽

Ulrike Bauder-Wüst ◽

Martin Schäfer ◽

Euy Sung Moon ◽

Klaus Kopka ◽

...

Keyword(s):

Human Serum ◽

Room Temperature ◽

Solid Phase ◽

Negative Influence ◽

Binding Motif ◽

Lncap Cells ◽

Cell Binding ◽

Binding Characteristics

Abstract Background The AAZTA chelator and in particular its bifunctional derivative AAZTA5 was recently investigated to demonstrate unique capabilities to complex diagnostic and therapeutic trivalent radiometals under mild conditions. This study presents a comparison of 68Ga, 44Sc and 177Lu-labeled AAZTA5-PSMA-617 with DOTA-PSMA-617 analogues. We evaluated the radiolabeling characteristics, in vitro stability of the radiolabeled compounds and evaluated their binding affinity and internalization behavior on LNCaP tumor cells in direct comparison to the radiolabeled DOTA-conjugated PSMA-617 analogs. Results AAZTA5 was synthesized in a five-step synthesis and coupled to the PSMA-617 backbone on solid phase. Radiochemical evaluation of AAZTA5-PSMA-617 with 68Ga, 44Sc and 177Lu achieved quantitative radiolabeling of > 99% after less than 5 min at room temperature. Stabilities against human serum, PBS buffer and EDTA and DTPA solutions were analyzed. While there was a small degradation of the 68Ga complex over 2 h in human serum, PBS and EDTA/DTPA, the 44Sc and 177Lu complexes were stable at 2 h and remained stable over 8 h and 1 day. For all three compounds, i.e. [natGa]Ga-AAZTA5-PSMA-617, [natSc]Sc-AAZTA5-PSMA-617 and [natLu]Lu-AAZTA5-PSMA-617, in vitro studies on PSMA-positive LNCaP cells were performed in direct comparison to radiolabeled DOTA-PSMA-617 yielding the corresponding inhibition constants (Ki). Ki values were in the range of 8–31 nM values which correspond with those of [natGa]Ga-DOTA-PSMA-617, [natSc]Sc-DOTA-PSMA-617 and [natLu]Lu-DOTA-PSMA-617, i.e. 5–7 nM, respectively. Internalization studies demonstrated cellular membrane to internalization ratios for the radiolabeled 68Ga, 44Sc and 177Lu-AAZTA5-PSMA-617 tracers (13–20%IA/106 cells) in the same range as the ones of the three radiolabeled DOTA-PSMA-617 tracers (17–20%IA/106 cells) in the same assay. Conclusions The AAZTA5-PSMA-617 structure proved fast and quantitative radiolabeling with all three radiometal complexes at room temperature, excellent stability with 44Sc, very high stability with 177Lu and medium stability with 68Ga in human serum, PBS and EDTA/DTPA solutions. All three AAZTA5-PSMA-617 tracers showed binding affinities and internalization ratios in LNCaP cells comparable with that of radiolabeled DOTA-PSMA-617 analogues. Therefore, the exchange of the chelator DOTA with AAZTA5 within the PSMA-617 binding motif has no negative influence on in vitro LNCaP cell binding characteristics. In combination with the faster and milder radiolabeling features, AAZTA5-PSMA-617 thus demonstrates promising potential for in vivo application for theranostics of prostate cancer.

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A role for fibroblast growth factor type-1 in nephrogenic repair. Autocrine expression in rat kidney proximal tubule epithelial cells in vitro and in the regenerating epithelium following nephrotoxic damage by S-(1,1,2,2-tetrafluoroethyl)-L-cysteine in vivo

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)50234-4 ◽

1993 ◽

Vol 268 (16) ◽

pp. 11542-11547

Author(s):

G. Zhang ◽

T. Ichimura ◽

J.A. Maier ◽

T. Maciag ◽

J.L. Stevens

Keyword(s):

Growth Factor ◽

Epithelial Cells ◽

Proximal Tubule ◽

Fibroblast Growth Factor ◽

Rat Kidney ◽

Fibroblast Growth ◽

Factor Type

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ErbB-2 Induces the Cyclin D1 Gene in Prostate Epithelial Cells In vitro and In vivo

Cancer Research ◽

10.1158/0008-5472.can-06-1898 ◽

2007 ◽

Vol 67 (9) ◽

pp. 4364-4372 ◽

Cited By ~ 27

Author(s):

Mathew Casimiro ◽

Olga Rodriguez ◽

Llana Pootrakul ◽

Maral Aventian ◽

Nadia Lushina ◽

...

Keyword(s):

Epithelial Cells ◽

Cyclin D1 ◽

Prostate Epithelial Cells

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In vitro and in vivo pathologic effects of Vibrio parahaemolyticus on human epithelial cells

Canadian Journal of Microbiology ◽

10.1139/m84-056 ◽

1984 ◽

Vol 30 (3) ◽

pp. 381-388 ◽

Cited By ~ 6

Author(s):

B. R. Merrell ◽

R. I. Walker ◽

S. W. Joseph

Keyword(s):

Epithelial Cells ◽

Epithelial Tissue ◽

Ileal Mucosa ◽

Cytotoxic Factor ◽

Culture Cells ◽

Vibrio Parahemolyticus ◽

Buccal Epithelial Cells ◽

Culture Supernate

The initial interaction and adherence of Vibrio parahemolyticus to epithelial tissue culture cells, human buccal epithelial cells, and the ileal mucosa of mice were studied. Using scanning electron microscopy, adherent bacteria were observed only on degenerating human embryonic intestinal, HeLa, and buccal cells; healthy normal cells were devoid of bacteria. Sheared V. parahaemolyticus, i.e., lacking flagella, did not adhere to either normal or degenerating tissue cells. Neither ultraviolet-inactivated organisms nor cell-free culture supernate affected the epithelial cells. Similar findings were observed on the mucosa of the ileum in mice inoculated with V. parahaemolyticus. It appears that V. parahaemolyticus possesses a cytotoxic factor which alters epithelial cells. This factor appears to be closely associated with viable organisms and may be a functional element in the adherence process of flagellated V. parahaemolyticus to mammalian epithelial cells.

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