Publication: Comparative histochemical and
biochemical studies on acid f3-galactosidase
activity in the experimentally injured rabbit cornea
and tear fluid using the sensitive substrate
f3-galactoside-4-trifluoromethylumbelliferyl (HFC)
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Date
1999
Authors
Cejkova, J. ; ZvBrova, J. ; Andonova, Z. ; Ardan, T.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
Comparative histochemical and biochemical
studies on acid B-galactosidase activity in the rabbit eye
after various experimental injuries were performed using
the same sensitive fluorogenic substrate B-galactoside-4-
trifluoromethylumbelliferyl (HFC). The aim of the study
was to examine whether the severity of corneal damage
corresponds with the level of the enzyme activity in the
tear fluid. As until recently the substrate B-galactoside-4-
HFC had not been used for the histochemical detection
of acid B-galactosidase in the cornea, results obtained
with this substrate in a fluorescent method were
compared in parallel cryostat sections with results
obtained using the substrate 5-bromo-4-chloro-3-indoxyl
B-galactoside in the indigogenic method (previously
shown to be very sensitive for the detection of acid Bgalactosidase
activity in the cornea). Both methods
revealed similar localization and changes in enzyme
activity; using B-galactoside-4-HFC an acceptable
cellular localization was achieved. For the measurement
of acid B-galactosidase activity in the tear fluid a
semiquantitative biochemical method was elaborated
using filter paper punches with the substrate (Bgalactoside-
4-HFC) soaked with tears and incubated at
37 "C. The time of the first appearance of a greenishyellow
fluorescence (enzyme positivity) was recorded by
UV lamp and compared with the appearance of
fluorescence in calibrated punches containing known
acid B-galactosidase activities. The results show that Bgalactoside-
4-HFC is useful for the biochemical
assessment of acid B-galactosidase activity in the tear
fluid. Comparing histochemical and biochemical results,
it can be concluded that increased enzymatic activity in
tears parallels the severity of corneal damage. Further
Offprint requests to: Prof. Jitka Cejkovh, MD, PhD, DSc, Head,
Department of Eye Histochemistry, Institute of Experimental Medicine,
Videhsak. 1083, 14220, Prague 4, Czech Republic
studies are necessary to evaluate whether the detection
of acid B-galactosidase activity in tears might be useful
for diagnostic purposes in humans.
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