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<title><string language="fre"><![CDATA[Dr Paul Williams - Calcium imaging of Cry5B action on the intestine of Ascaris suum]]></string></title>
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<string language="fre"><![CDATA[Crystal (Cry)
proteins from Bacillus thuringiensis are
used as effective insecticides. These pore forming toxins act within the midgut
of insects killing them. Cry5B, has been found to target and kill intestinal
parasitic nematodes including Ascaris
suum. Cry5B forms pores in the nematode intestines killing the parasite in
less than a day. To track the time course of the action of Cry5B on the
intestine beginning from application, we incubated Ascaris suum intestines in Fluo-3AM for calcium imaging. We exposed
isolated intestine flaps to either 100 µg/ml or 10 µg/ml activated Cry5B and
followed the cytosolic  calcium over a
6-hour period. We observed a Cry5B concentration-dependent increase and
subsequent fall in calcium that occurred after 1 hour from Cry5B application.
The shortest time that we observed an increase in calcium after application of
Cry5B was 10 minutes. After the initial increase in calcium, we observed small
fluctuations and oscillations in the calcium signal. We are treating samples
with 100 mM galactose to see if there is inhibition of the effects of Cry5B and
histology to look for the microscopic structure changes. By 6 hours intestinal
cells were severely damaged, with their columnar cells vacuolated and swollen.
The activated Cry5B toxin can act rapidly to produce serious nematode intestine
damage. With PCR and qPCR techniques we have identified the presence of BRE-5
and other Cry5B target orthologues showing intestine specific expression
present in Ascaris suum. The complex
calcium signal requires further study.
 Financial Support: RJM is supported by NIH, the National Institute of
Allergy and Infectious Diseases grants R01AI047194-17, R21AI092185-01A1
and the E. A. Benbrook Foundation for Pathology and
Parasitology.]]></string></description>
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<date><dateTime>2021-11-29</dateTime></date>
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