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Yeast
cell
The
Delayed Luminescence (DL) of temperature sensitive Saccharomyces
Cerevisiae CDC 28-1 was measured in two different conditions: yeast
that has been maintained at the restrictive temperature of 38.5 °C
for some hours (and for this should be at the same step of the
division cycle ) and samples that remained always at 23 °C. In both
cases DL shows a typical hyperbolic trend but the DL parameters
appears to be different according the previous history of the
culture. The connection between the DL parameters and some
parameters of the culture (thermal treatment, glucose contents of
extracellular liquid, density and age of the culture) has been
evinced.
Ionizing
radiation
In
the framework of the interaction between the ionizing radiation and
the biological systems, the analysis of the ultraweak luminescence
could give information about the answer of the living systems to the
radiation. In particular the analysis of the ultraweak luminescence,
that gives very sensitive indication of the functional state of the
biological system, could be particularly useful to study the effect
of the ionizing radiation low doses. The measurement of photoinduced
luminescence has been carried on biological samples that has been
irradiated by gamma rays, X rays and UV, at doses where it has been
possible to measure a change of the biological behaviour of the
samples. In collaboration with other researchers from Modena
University (I) it has been made the measurement of delayed
luminescence from yeast cell irradiated by soft X-rays. The total
remitted radiation shows for this experiment a monotonic decrease vs
the X-rays dose. The above results have been obtained for relatively
high doses and have shown that also in the case of damage induced by
ionizing radiation it is possible to discriminate the effects by
means of the ultraweak luminescence analysis. Besides, once this
correlation at high doses has been experimentally consolidated, one
should try to make the experiments at low doses where the
peculiarity of the proposed analysis is more suitable; in fact, at
low doses, the biological interested mechanisms from the ionizing
radiation seem to be attributed to a different cellular answer and
for this reason the ultraweak luminescence should be sensitive
enough, since it reflects the behaviour of the whole system, that is
strictly connected to the behaviour of the cellular mechanism.
Acetabularia
In
order to study the connection between stress condition and DL
behaviour, several experiments are performed on Acetabularia
Acetabulum (AA); in particular we are studying:
a)
The dependence of the emission spectrum on the
temperature.
These
spectra show a very weak dependence on the temperature; it appears
that, on decreasing the temperature the spectrum moves towards
higher frequencies while a measurable change of the form is not
noticed.
b)
The dependence of the dynamics on the wavelength and on the
intensity of the source.
There
is a small but clear change in the slope of the curves dependent on
the excitation wavelength. On the other hand there is a strong
dependence of the slope on the intensity of illumination: in fact on
increasing the intensity of the illumination source the slope of the
curves increases up to a saturation value and then the slope begins
to decrease while characteristic oscillations appears. Another
aspect of the problems connected to the interpretation of the
dependence of the dynamics on the intensity of the source is the
fact that the same AA sample illuminated with light of the same
frequency and with the same times of illumination but with different
intensities present decay curves that often cross each other. This
fact is incompatible with a simple description of the phenomenon
that involves the population and the following depletion of
electronic levels. |
c)
The dependence of the dynamics on the temperature.
As
it concerns the dependence of the dynamics on the temperature the
existence of two regimes is evident: in the first regime, which is
the range of temperature within which the AA could survive, it is
possible to bring the AA to the initial conditions again , even if
not immediately but following a kind of cycle; instead in the second
regime, characterised by excessively high temperatures, the DL
emission from the AA decreases in a irreversible way and after some
time the sample dies. This decreasing of the DL emission probably is
connected to the damages that the extreme temperatures causes on the
functional structure of the AA. In fact even if, after a rapid
freezing in liquid air, an AA sample is incubated at its normal
temperature, the DL emission practically disappears as in the
samples submitted to high temperatures.
d)
The dependence of the dynamics on some specific ions.
It
seems that the presence of Ca++ seems to produce a strong decrement
both of the total number of excited levels and of the decay
probability. In fact if it is present only Na+ and Ca++ the total
number of excited levels go down very speedily (reaching 10% of the
initial value in about 1 hour) and the AA appear strongly damaged.
The presence of others cations K+ and Mg++ seems to reduce the
effect of Ca++. In fact if they are presents together the AA lives a
long time and its DL is quite constant. Moreover if only they are
present there is no big effect on the behaviour of DL at least in
the first hours. This fact is confirmed also if only K+ is present
for the first 4 hours. On the contrary the presence of Mg++ produce
strong changes of DL. Moreover it is shown clearly that the
decreasing of the absolute value of the membrane potential causes a
quenching of the DL with a diminution of the total number of excited
centres and an increase of the decay probability.
e)
DL and cellular organization.
A
microscopic procedure was used to register massive intracellular
movements in of Acetabularia cells. Results of "simultaneous"
measurements of chloroplast streaming and delayed luminescence of
Acetabularia cells showed that when the chloroplasts movement was
inhibited by rapid freezing in liquid nitrogen or incubation in
chloroform drastic changes in DL occur too. It is known that
chloroplast movement is connected to the underling cytoskeleton. In
addition the link between DL and cellular organization appeared also
evident in an other study where the effect of cations in the
extracellular medium on the parameters of DL was compared to
morphological changes in Acetabularia cells. Calcium ion influx has
indeed been shown to cause rearrangement of the underlying
cytoskeleton in Acetabularia. Results suggest the idea that delayed
luminescence is related to the integrity of the dynamic chloroplast
organization, even if both the relation between chloroplast
streaming and DL has not been yet unambiguously determined and the
source of such a relation is still not clear. Further studies could
give an explanation of such a connection, thus DL could be used as
experimental technique to investigate and register the integrity of
the massive intracellular movement.

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