Differential expression of cyclophilin isoforms during keratinocyte differentiation

Cyclophilin A, the major intracellular binding protein for the immunosuppressive drug cyclosporin A (CsA), was studied in human keratinocytes during differentiation both in vivo and in vitro. Analysis of cyclophilin by gel-filtration radiobinding-assay with tritiated CsA showed one specific radioactive peak at 17 kDa. By this technique, the levels of cyclophilin (mean 55.23 +/- 8.43 pmol/mg protein) did not significantly differ during keratinocyte differentiation. When the protein extracts from calcium-induced differentiating keratinocytes and normal human skin were analysed by PAGE radiobinding-assay, two specific radioactive CsA-binding peaks were detected. The major peak (RF 0.13) was expressed in all samples (mean 47.32 +/- 17.53 pmol/mg protein) whereas the minor peak (RF 0.23) was dramatically decreased about 6-fold in abnormally differentiated skin (psoriasis) as well as in non-differentiated keratinocytes. At least six [3H]CsA-binding isoforms with pI values ranging from 5.58 to 7.75 were detected by isoelectrofocusing autoradio-blotting-assay in normal human skin; three of them immunoreacted with antibodies to cyclophilin. These results demonstrated the presence of several cyclophilin isoforms in human epidermal cells and an expression which correlated with the differentiation of human keratinocytes both in vivo and in vitro.

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Reconstructed Human Epidermis In Vitro: An Alternative to Animal Testing

Alternatives to Laboratory Animals ◽

10.1177/026119299502300113 ◽

1995 ◽

Vol 23 (1) ◽

pp. 97-110 ◽

Cited By ~ 1

Author(s):

Maria Ponec

Keyword(s):

Human Skin ◽

Three Dimensional ◽

Structural Features ◽

Animal Testing ◽

Normal Human Skin ◽

Normal Human ◽

Biochemical Mechanisms ◽

Human Skin Models

Various three-dimensional human skin models, in which the epidermis exhibits in vivo-like morphological and functional characteristics, have recently been developed. Such models are currently being used to study the development and physiology of the skin, the processes involved in wound healing, and the reactivity of skin to environmental and chemical insults. Since these models reproduce to a large extent the barrier function properties of normal human skin, they can be used for screening potential skin irritants. These substances can be applied topically and their irritant potential can be evaluated using various endpoints, such as the induction of tissue damage or the release of various pro-inflammatory mediators. Studies with human skin equivalents can therefore contribute to our knowledge of the basic biochemical mechanisms underlying irritant reactions, and can be used to understand the structural features of molecules which may be responsible for eliciting an irritant reaction. In addition”, the generation of epidermal equivalents populated with melanocytes, as well as keratinocytes, makes it possible to study the regulation of melanogenesis, melanocyte–keratinocyte interactions, and how these are affected by UV irradiation. Such a model can also be used for testing the phototoxic or photoprotective potentials of various compounds and sunscreens.

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The Effect of Practolol, In Vitro Generated Metabolites of Practolol and Chemical Analogues on the Rate of Growth of Human Skin Fibroblasts

Alternatives to Laboratory Animals ◽

10.1177/026119298401200205 ◽

1984 ◽

Vol 12 (2) ◽

pp. 89-97

Author(s):

Graham R. Elliott ◽

H.E. Amos ◽

James W. Bridges

Keyword(s):

Human Skin ◽

Psoriasis Patient ◽

Skin Fibroblasts ◽

Human Skin Fibroblasts ◽

Cell Type ◽

Normal Human Skin ◽

Rate Of Growth ◽

Structural Analogues ◽

Normal Human

The rate of growth of normal human skin fibroblasts was inhibited in a dose related, reversible, fashion by practolol (N-4-(2-hydroxy)-3 (1-methyl)-aminopropoxyphenylacetamine) (ID50 1.35 ± 0.14 x 10-3M), propranolol (1-(isopropylamino)-3(1-naphthyl-oxy)-2-propranolol) (ID50 0.145 ± 0.02 x 10-3M) and paracetamol (N-(4-hydroxyphenyl) acetamide) (ID50 0.85 ± 0.2 x 10-3M). Skin fibroblasts isolated from a psoriasis patient were more sensitive towards practolol (ID50 0.48 ± 0.14 x 10-3M) and propranolol (ID50 0.032 ± 0.002 x 10-3M), but less sensitive towards paracetamol (ID50 1.3 ± 0.07 x 10-3M). In vitro generated metabolites of practolol, using normal or Arochlor 1254-pretreated hamster liver preparations, and structural analogues of practolol had no effect upon the growth of either cell type.

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Differential effects of 5-aminolaevulinic acid photodynamic therapy and psoralen + ultraviolet A therapy on p53 phosphorylation in normal human skin in vivo

British Journal of Dermatology ◽

10.1111/j.1365-2133.2005.06922.x ◽

2005 ◽

Vol 153 (5) ◽

pp. 1001-1010 ◽

Cited By ~ 22

Author(s):

L.E. Finlan ◽

N.M. Kernohan ◽

G. Thomson ◽

P.E. Beattie ◽

T.R. Hupp ◽

...

Keyword(s):

Photodynamic Therapy ◽

Human Skin ◽

Ultraviolet A ◽

Normal Human Skin ◽

Differential Effects ◽

Aminolaevulinic Acid ◽

Normal Human

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AUTOCRINE-ACTING EARLY SECRETED MITOGENIC ACTIVITY: PRODUCTION AND RESPONSIVENESS IN CULTURES OF NORMAL HUMAN KERATINOCYTES AS A FUNCTION OF IN VIVO AND IN VITRO AGE

Cell Biology International ◽

10.1006/cbir.2000.0624 ◽

2001 ◽

Vol 25 (3) ◽

pp. 197-204 ◽

Cited By ~ 2

Author(s):

M Tenchini

Keyword(s):

Human Keratinocytes ◽

Mitogenic Activity ◽

Normal Human Keratinocytes ◽

Normal Human

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The Effect of In Vivo Interferon-Gamma on the Distribution of LFA-1 and ICAM-1 in Normal Human Skin

Journal of Investigative Dermatology ◽

10.1111/1523-1747.ep12284016 ◽

1989 ◽

Vol 93 (4) ◽

pp. 439-442 ◽

Cited By ~ 86

Author(s):

Jonathan N.W.N. Barker ◽

Michael H Allen ◽

Donald M MacDonald

Keyword(s):

Human Skin ◽

Interferon Gamma ◽

Normal Human Skin ◽

Normal Human

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Galanin receptor expression in cultured human keratinocytes and in normal human skin

Journal of the Peripheral Nervous System ◽

10.1111/j.1085-9489.2006.00081.x ◽

2006 ◽

Vol 11 (2) ◽

pp. 156-164 ◽

Cited By ~ 19

Author(s):

Attila Dallos ◽

Maria Kiss ◽

Hilda Polyanka ◽

Attila Dobozy ◽

Lajos Kemeny ◽

...

Keyword(s):

Human Skin ◽

Human Keratinocytes ◽

Receptor Expression ◽

Galanin Receptor ◽

Normal Human Skin ◽

Normal Human

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Identification of a trypanocidal factor againstTrypanosoma equiperdumin normal human serum

Parasitology ◽

10.1017/s0031182000061485 ◽

1989 ◽

Vol 98 (3) ◽

pp. 401-407 ◽

Cited By ~ 11

Author(s):

G. Verducci ◽

S. Perito ◽

R. Rossi ◽

E. Mannarino ◽

F. Bistoni ◽

...

Keyword(s):

Human Serum ◽

Gel Filtration ◽

Density Lipoprotein ◽

Natural Antibodies ◽

Normal Human Serum ◽

Mouse Serum ◽

Trypanocidal Activity ◽

Normal Human

SUMMARYNormal human serum (HS) contains trypanolytic activity and agglutinins toTrypanosoma equiperdum, while such activities are not found in sera from a range of animals susceptible to infection. HS given toT. equiperdum-infected mice caused a rapid decrease in the number of circulating trypanosomes and protection from lethal infection. Trypanolytic activity of human serum was found to be associated, after DEAE chromatography and Sephadex G-200 gel filtration, with the fraction containing 19S antibodies. Immunofluorescence assays confirmed a binding of human IgM and C1qcomplement component onto the surface ofT. equiperdum. Anti-T. equiperdumactivity of HS was specifically directed toT. equiperdumsurface components and not to some mouse serum components adsorbed on parasites during the growth in the host, because HS adsorbedin vivoin CD-1 mice retained full protective and agglutinating properties. Trypanocidal activity appears in human serum about the 7th month after birth and persists until late in life. On the contrary, human purified high-density lipoprotein had no significantin vitroorin vivotrypanocidal activity. In conclusion, strong natural anti-T. equiperdumactivity in human serum was mainly mediated by natural antibodies of the IgM class. The presence of natural IgM active againstT. equiperdumin HS could represent one of the natural mechanisms of resistance of refractory hosts against trypanosome infections. This phenomenon provides further evidence that host specificity of trypanosomes may be partly conditioned by the presence of natural antibodies.

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Synthesis and In Vitro Anticancer Activity of 6-Ferrocenylpyrimidin-4(3H)-one Derivatives

Current Organic Synthesis ◽

10.2174/1570179415666181113143516 ◽

2019 ◽

Vol 16 (1) ◽

pp. 160-164 ◽

Cited By ~ 6

Author(s):

Stanislav A. Grabovskiy ◽

Rinat S. Muhammadiev ◽

Lenar R. Valiullin ◽

Ivan S. Raginov ◽

Natalie N. Kabal'nova

Keyword(s):

Anticancer Activity ◽

Anticancer Agents ◽

Practical Interest ◽

Breast Adenocarcinoma ◽

Human Skin Fibroblast ◽

Normal Human Skin ◽

Normal Human ◽

Mcf 7

Aim and Objective: Some ferrocenyl derivatives are active in vitro and in vivo against cancer. Generally, ferrocenyl derivatives for cancer research have three key components: a ferrocene moiety, a conjugated linker that lowers the oxidation potential and some derivative (peptide, nucleobase and others) that can interact with biomolecules. Since the pyrimidine fragment can easily pass through the membrane into the cells and become involved in metabolism; it appears to be promising. Furthermore, this fragment is an electron-acceptor group, so a spacer can be excluded. Therefore, the synthesis of 6-ferrocenylpyrimidin-4(3H)-one derivatives and the study of their anticancer activity have scientific and practical interest. </P><P> Methods: The syntheses of 6-ferrocenylpyrimidin-4(3H)-one derivatives were performed by the condensation of ethyl 3-ferrocenyl-3-oxopropionate with thiourea or acetamidine or guanidine. The cytotoxicity of four 6- ferrocenylpyrimidin-4(3H)-one derivatives was evaluated by using the MTT assay in vitro against Human breast adenocarcinoma MCF-7 and normal human skin fibroblast HSF cells. The tested derivatives induced a concentration-dependent cytotoxic response in cell lines. </P><P> Results: A study of the cytotoxic activity of 6-ferrocenylpyrimidin-4(3H)-one derivatives by the MTT test has found that all compounds have a dose-dependent toxic effect on the lines of breast cancer cells (MCF-7) and normal human fibroblast cells (HSF). The most pronounced cytotoxic effect is exhibited by 2-methyl-6-ferrocenylpyrimidin- 4(3H)-one (MCF-7, IC50 17 ± 1 µM). Conclusion: The experimental results confirm the importance of investigation and design of ferrocenylpyrimidin- 4(3H)-one derivatives as anticancer agents. Compounds where the pyrimidine derivatives are directly linked to the ferrocene unit rather than via a spacer group also may be of interest for antiproliferative drug design.

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