Acrylamide (AA) is one of the most common toxins in foods. Its
effect on bone microstructure has not been investigated. The aim
of our study was to analyze the impact of acute exposure to AA on
femoral bone microstructure in mice. Adult animals were treated
perorally with 2 doses of AA (E1 group, 1 mg/kg b.w.) in a 24-h
period and with 3 doses of AA (E2 group, 1 mg/kg b.w.) in a 48-h
period. Mice exposed to AA had smaller sizes of primary osteon's
vascular canals. Secondary osteons were significantly smaller in
mice from E2 group; however their increased number (from 38 %
to 77 %) was identified in both E1 and E2 groups. In these groups,
a higher number of resorption lacunae (from 100 % to 122 %) was
also found. The values for bone volume, trabecular number were
increased and that for trabecular separation was decreased in mice
administered AA. Significantly higher value of bone surface was
observed in mice from E1 group whereas trabecular thickness was
increased in E2 group. The effect of AA on microstructure of
compact and trabecular bone tissues is different. In our study, one
dose of AA was used and acute effects of AA were investigated.
Therefore, further studies are needed to study mechanisms by
which AA acts on bone.
Early mouse neural stem cells (NSCs) first appear in embryonic day E5.5 and express pluripotency markers Oct4, Sox2, Nanog and early neural marker Sox1. Early NSCs are a good model for understanding the role of various pathways that control initial neural commitment. However, a protocol for differentiation of mouse embryonic stem cells (ESCs) into early NSCs by adherent monolayer culture has not yet been established. Hence, in this study, we identified the combination of growth factors and small molecules that differentiated mouse ESCs into early NSCs and supported their proliferation. Leukaemia inhibitory factor (LIF) was the first factor to be tested and it was found that ESCs can differentiate into early neurogenic lineage in the presence of LIF. However, we found that the induction is weaker in the presence of LIF as compared to cells differentiated in its absence. GSK-3 inhibitor, along with BMP and TGF-β pathway inhibitor (dual SMAD inhibition), are commonly used to sequentially direct ESCs towards NSCs. However, when we used this combination, mouse ESCs failed to differentiate into early NSCs. We observed that by adding Wnt inhibitor to the combination of GSK-3 inhibitor, BMP inhibitor, TGF-β inhibitor and LIF, it was possible to differentiate ESCs into early NSCs. qRT-PCR analysis of early NSCs illustrated that they expressed key pluripotency genes Oct4 and Nanog, albeit at levels lower than non-differentiated ESCs, along with early neural markers Sox1 and Pax6.
Taurine, a sulphur - containing amino acid, has been termed
a functional nutrient. Its synthetic form is a common ingredient
in supplements and energy drinks. There is no information
concerning taurine impact on bone microstructure after
prolonged supplemental use. Also, differences in bone
parameters of mice following taurine exposure are unknown. In
this study, a detailed microstructure of compact and trabecular
bone tissues of mice subchronically exposed to taurine was
determined. Animals (n=12) were segregated into three groups:
E1 group – mice received 20 mg/kg b.w. of taurine per day
during 8 weeks; E2 group – mice were fed by taurine at a dose
of 40 mg/kg b.w. for 8 weeks and a control (C) group. Decreased
density of secondary osteons, increased sizes of primary osteon's
vascular canals (P<0.05) were observed in taurine – treated
animals. Cortical bone thickness, trabecular thickness were
decreased (P<0.05) in E1 group, and relative volume of
trabecular bone was lower (P<0.05) in E2 group as compared to
C group. According to our results, prolonged taurine exposure at
the doses used in this study can negatively affect both compact
and trabecular bone tissues microstructure.
The study examined the morphological and long-term behavioral impacts of neonatal hypoxic-ischemic brain injury in a mouse model. We investigated the modification of different behavioral domains, such as spontaneous climbing, which represents fine motor skills. We also focused on sex-dependent differences during hypoxic-ischemic encephalopathy. The Rice-Vannucci model of hypoxia-ischemia was used, adjusted and adapted to 7-day-old C57BL/6NTac mice. The effects of induced hypoxia and ischemia were also studied separately. At postnatal day 60, mice underwent behavioral testing using the LABORAS apparatus. The perfusion for histological evaluation was performed one day after the behavioral analyses. In groups with separately induced hypoxia or ischemia, the observed alterations in behavior were not accompanied by morphological changes in the cortex or hippocampal formation. Female mice naturally climbed significantly more and hypoxic females reared less than hypoxic males (p<0.05). Male mice postnatally exposed to hypoxiaischemia exhibited significantly lower vertical activity and higher horizontal activity (p<0.05). Mild hypoxic damage may not be morphologically detectable but may induce substantial behavioral changes in adult mice. There were significant differences between horizontal and vertical activity in reaction to hypoxiaischemia. Our study indicates that the importance of behavioral testing is irreplaceable and may be reflected in neonatal medicine.
Optical mapping is a fluorescence-based physiological method to image spreading of action potential in excitable tissues, such as the heart and central nervous system. Because of the requirements for high speed imaging in low light conditions, highly sensitive high-speed cameras together with an optical system with maximum photon efficiency are required. While the optimization of these two components is relatively straightforward, the choice of the perfect light source is less simple; depending on the other (usually fixed) components, various parameters may acquire different weight in decisionmaking process. Here we describe the rationale for building an optical mapping setup and consider the relative advantages and disadvantages of three different commonly available light sources: mercury vapor lamp (HBO), xenon lamp (XBO), and light emitting diode (LED). Using the same optical system (fluorescence macroscope) and high-speed camera (Ultima L), we have tested each of the sources for its ability to provide bright and even illumination of the field of view and measured its temporal fluctuations in intensity. Then we used each in the actual optical mapping experiment using isolated, perfused adult mouse heart or chick embryonic heart to determine the actual signal to noise ratio at various acquisition rates. While the LED sources have undergone significant improvements in the recent past, the other alternatives may still surpass them in some parameters, so LEDs may not be the automatic number one choice for every application., Veronika Olejnickova, David Sedmera., and Obsahuje bibliografii