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Intl. J. of Molecular Zoology, 2012, Vol.2, No.1, 1
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accordance with the findings of (Lock et al., 1999)
who demonstrated that tTG was distributed in both
rodent and primate tissues identically with the
extracellular reticulin-endomysial-jejunal staining
pattern of sera from Coeliac patients. Ivarsson (2005)
suggest that CD may be prevented by improving early
feeding. Since that time more infants were still
breastfed when gluten was introduced in smaller
amounts and this was followed by a sharp decline of
the previous incidence level of CD in children. These
findings open the way to possible prevention
strategies, by introducing the adequate quantity of
gluten at the optimal time, during the period of
breastfeeding. In our study we compare and clarify the
role of early bovine colostrums or (ongoing)
breastfeeding with gluten introduction in the
prevention of the development of CD. Breast feeding
in our study never prevented CD crisis when gluten
was introduced till weaning in group 1 suggesting the
passage of gluten peptide to the lamina propria that
cause inflammation in the gut and intestinal mucosa
due to increased level of tTG in this group as
confirmed by scanning electron microscope. Another
possible explanation that the introduction of gliadin
with different doses concomitant with interferon-γ
abrogated the incidence level of Coeliac disease that
may interfere with milk composition with respect to
the duration of breastfeeding in our experiment. In
addition, in rodents, two growth factors play an
important role in the process of regeneration and
proliferation of enterocytes: epidermal growth factor
(EGF) and transforming growth factor (TGF)-α.
However, TGF-α has not been found in rat maternal
milk (Dvorak et al., 1994). In our investigation, the
scanning electron micrographs in group 1 showed
deformed villi in some areas while others exhibited
sloughing in their tips due to the increased level of
tTG. Thus, it is likely that the increased introduction to
gluten in small intestine could affect the synthesis of
EGF and TGF-α that may enhance the deficit of
maternal milk to protect against coeliac crisis.
The content of immunoglobulins in colostrum and
milk is highly dependent on the animal species (Butler
et al., 2005). Thus, for many species the proportion of
IgA increases between colostrum and milk (Mix et al.,
2006). For animals like rats, mice, dogs and ungulates,
uptake of colostrum of adequate quality and sufficient
quantity is important for the offspring to boost the
systemic immune function in the short term. Immune
factors from bovine colostrum supplementation are
not digested and absorbed, but remain intact and
active in the intestinal tract. As such, colostrum
participates in gutassociated lymphoid tissue (GALT)
activity, and therefore plays a role in both immune
health and gastrointestinal function (Mix et al., 2006).
Notably, recent studies have suggested the occurrence
of an innate immune response to gluten in Coeliac
patients (Meresse et al., 2004; Maiuri et al., 2005).
Furthermore, the requirement for calcium activation of
the tTG enzyme has been disputed. Our finding that
calcium is not essential is in agreement with recent
reports (Haroon et al., 1999). In our investigation,
pups fed bovine colostrums were significantly
improved than suckling pups when assessed by their
body weight. The mean body weight measured on
group 2 samples were the highest by the end of
weaning (32.41±1.75) and significantly differ from
those observed on the other days) among the other
groups of pups. In summary, our data for the first time
suggest that the uptake of bovine colostrum could
minimize the severity of GI lesions in coeliac disease
concomitant with autism.
3. Materials and Methods
3.1 The valproic acid rat model of autism and
Coeliac disease
Female Wistar rats with controlled fertility cycle were
mated overnight and the morning when spermatozoa
were found was designated as the first day of gestation.
Females were on a standard diet and received a single
intraperitoneal injection of 600 mg/kg sodium
valproate on day 12.5 after conception. Control
females were kept on normal standard diet and
injected with physiological saline at the same time.
Sodium valproate (Sigma) was dissolved in saline at a
concentration of 250 mg/mL. Administration of this
dose to rats during embryogenesis has been shown to
result in a maximum level of total VPA (900 μg/mL)
in maternal plasma in less than 1 h, with a mean
plasma elimination half-life of 2.3 h (Binkerd et al.,
1988). Dams were housed individually and were
allowed to raise their own litters. 1
st
and 2
nd
experimental groups were treated with interferon-γ
(1 000 U per animal, administered intraperitoneally)
after birth. Gliadin (0.5 and 3 mg) was intragastrically
administered to the pups of the 1
st
and 2
nd
groups on
days 0 and 3, and a 30 mg challenge dose was given
on day 20 (24 hours before the termination of the
experiment) (Štěpánková et al., 2003).