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APRIL 24
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– 26
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, 2014 | OLOMOUC | THE CZECH REPUBLIC
ABS TRAC T BOOK
Studies have shown that there is variation in the agreement
between operators viewing the same tissue [1] suggesting that
a complimentary technique for verification could improve the
robustness of the evaluation, and improve patient care. FTIR
chemical imaging allows the spatial distribution of chemistry to
be rapidly imaged at a high (diffraction limited) spatial resoluti-
on where a pixel represents an area of 5.5 × 5.5 µm
2
of tissue. At
each pixel there is a full infrared spectrum providing a chemical
fingerprint which studies have shown contains the diagnostic
potential to discriminate between different cell types, and even
the benign or malignant state of prostatic epithelial cells [2-4].
We report a label-free (
i.e.
no chemical de-waxing, or staining)
method of imaging large pieces of prostate tissue (typically
1 cm × 2 cm) in tens of minutes yielding images containing
millions of spectra. Spectra are then automatically classified as
one of seven cell-types in prostate tissue in a matter of seconds.
This can be extended to classify the tissue as cancerous or
non-cancerous and even predict the grade of the cancer [5].
Similar methodology has been used to assess breast cancer
sample in tissue micro arrays [6].
References
1.
Allsbrook WC, et al. Interobserver reproducibility of Gleason grading of prostatic car-
cinoma: Urologic pathologists. Human Pathology, 2001; 32(1): 74–80.
2.
Fernandez DC, et al. Infrared spectroscopic imaging for histopathologic recogniti-
on. Nature Biotechnology, 2005; 23(4): 469–474.
3.
Baker MJ, et al. FTIR-based spectroscopic analysis in the identification of clinically
aggressive prostate cancer. British Journal of Cancer, 2008; 99(11): 1859–1866.
4.
Gazi E, et al. A correlation of FTIR spectra derived from prostate cancer biopsies with
Gleason grade and tumour stage. European Urology, 2006; 50(4): 750–761.
5.
Bassan P, et al. Automated high-throughput assessment of prostate biopsy tissue using
infrared spectroscopic chemical imaging, Proceedings of SPIE Accepted.
6.
Bassan P, et al. The potential use of glass substrates for FTIR chemical imaging of ti-
ssue microarrays (TMA), Anal. Chem. 2014; 86(3): 1648–1653.
The new era of glycohistochemistry: how human
lectins translate the sugar code
Gabius H. J.
Chair of Physiological Chemistry, Faculty of Veterinary
Medicine, Ludwig-Maximilians-University Munich,
Germany
Histochemists are familiar with the occurrence of glyco-
sylation and the presence of cellular glycoconjugates. In fact,
glycan structures detected by laboratory tools (plant/inverte-
brate lectins, monoclonal antibodies) can be markers for cell
typing or an activation status, their profile serving as molecular
fingerprints. The growing awareness of the unsurpassed coding
capacity of oligosaccharides and the role of human lectins
to translate this information into cellular effects give us an
entirely fresh view on the significance of glycosylation (1, 2).
Following an introduction to the concept of the sugar code,
case studies on physiological orchestration of glycosylation/
lectin expression in tumor-suppressor-dependent tumor cell
growth regulation and in autoimmunity will guide to under-
stand the biofunctionality of glycan-lectin recognition (for
review, please see 3). A brief survey of the toolbox, applied
together with Prof. K. Smetana Jr. (Charles University), is then
intended to inspire joint research projects.
References
1.
H.-J. Gabius (ed) The Sugar Code. Fundamentals of glycosciences. Wiley-VCH, 2009.
2.
H.-J. Gabius et al., From lectin structure to functional glycomics: the sugar code.
Trends Biochem. Sci. 2011; 36: 298–313.
3.
K. Smetana Jr. et al., Context-dependent multifunctionality of galectin-1: a challenge
for defining the lectin as therapeutic target. Exp. Opin. Ther. Targ. 2013; 17: 379–392.
The emerging significance of the tumor
microenvironment: examples on importance of
cell contacts and effector secretion
Kodet O.
1, 2
, Dvořánková B.
1
, Szabo P.
1
, Gabius H.-J.
3
,
Krejčí E.
1
, Strnad H.
4
, Kolář M.
4
, Lacina L.
5
, GrimM.
1
,
Dvořák P.
6
, Smetana K. Jr.
1
1
Charles University, 1
st
Faculty of Medicine, Institute
of Anatomy, Prague, Czech Republic
2
Charles University, 1
st
Faculty of Medicine, Department
of Dermatovenereology, Prague, Czech Republic
3
Ludwig-Maximilians University, Faculty of Veterinary Medi-
cine, Institute of Physiological Chemistry, Munich, Germany
4
Institute of Molecular Genetics, Academy of Science of
the Czech Republic vvi, Prague, Czech Republic
5
Institute of Medical Biology, A*STAR, Singapore
6
Institute of Medical Biology, Faculty of Medicine,
Masaryk University, Brno, Czech Republic
The increasing incidence of malignant melanoma andmorta-
lity prompt intense efforts to study tumor biology. Themalignant
cells of melanomas derive from normal melanocytes, mela-
nocytes themselves developing from neural crest-originating
precursors. These stem cells with capacity to generate new
melanocytes remain in the bulge region of hair follicles up to
the adult age. We here direct attention to interactions between
tumor cells and cells in their microenvironment. Of interest,
the early embryonic microenvironment is able to reverse the
aggressive behavior of malignant cells isolated from advanced
melanomas, cancer cells migrating and docking at sites typical
for neural crest cells. Conditionedmedia fromcultures of human
embryonic stem cells and cancer-associated/normal fibroblasts
are able to influence the phenotype of cells prepared from
advanced tumors. Further cell types occurring in themelanoma
niche are keratinocytes. The epidermis overlaying the nodular
melanoma has a pseudohyperplastic character with aberrant
expression of keratins but with a reduced proliferation activity.
Malignant melanocytes or neural crest stem cells significantly
influence the differentiation pattern of cocultured keratinocytes
and partially reduce their proliferation activity. Potent effector
proteins such as IL-8, CXCL-1, FGF-2, and VEGFA appear to play
a role in the keratinocyte-melanoma cells crosstalk. Such an in-
tercellular interplay in cancer may have a rather general character.
In respective work, we previously observed that melanoma-asso-
ciated fibroblasts are bioactive on breast cancer cells. Moreover,
