Ana Katušić

The effect of vibration therapy on spasticity and motor function in children with cerebral palsy: A randomized controlled trial

As the motor system relies heavily on deep sensory stimulation, recent studies have investigated the effect of vibration stimuli. Although research suggests a positive influence of vibration on motor performance in individuals with neurological disorders, there are very limited numbers of studies in children with cerebral palsy (CP). The objective of the present study was to evaluate the effects of sound wave vibration therapy on spasticity and motor function in children with CP. In this 3-month trial, 89 children with spastic CP were randomized to either continue their physiotherapy treatment (PT) or to receive vibration therapy twice a week in addition to their PT program. The randomization was stratified according to the Gross Motor Function Classification System (GMFCS) level to ensure similar functional ability. Children were assessed at baseline and after the 12-week intervention period. The outcomes measured were spasticity level as assessed by Modified Modified Ashworth Scale (MMAS) and gross motor function as assessed by Gross Motor Function Measurement (GMFM-88). Subgroup analysis was performed for the GMFCS. Significant differences between groups were detected for changes in spasticity level and gross motor function after the three months intervention. In conclusion, vibration therapy may decrease spasticity and improve motor performance in children with CP. The results of the present trial serve as valuable input for evidence-based treatments in paediatric neurorehabilitation.

Visual stimulations' critical period in infants with perinatal brain damage.

BACKGROUND Children with perinatal brain damage have a high prevalence of visual impairment. Stimulation of vision at a critical period can encourage brain plasticity and the recovery of impaired function. OBJECTIVE The aim of our study was to investigate when is the critical period for visual stimulation in children with perinatal brain damage. METHODS We compared 35 children within the first eight months of life (median age = 4 months) to 35 children aged between eight and thirty months (median age = 15 months), all with perinatal brain damage. All the children were attending an early intervention program at Mali dom, Zagreb, a rehabilitation centre for children with visual impairment. We compared the results from baseline and follow-up assessments of visual functions (grating acuity and contrast sensitivity). We also compared differences in change scores between the two groups. RESULTS Our results have shown that children who commence a visual stimulation program within the first eight months of life had more improvement in both visual functions. This improvement is statistically significant in visual acuity (p = 0.048). CONCLUSIONS These results indicate the importance of starting a visual stimulation program within the first eight months after birth.

PO-362 Influence of epigenetic agents on mouse teratoma development in vitro

Introduction Testicular Germ Cell Tumours (TGCT) are the most frequent malignancies in young male population and believed to be initiated by epimutations, i.e. aberrant epigenetics. Among various, teratoma is the most differentiated TGCT type encompassing all three germ layer derived tissues. Mouse teratoma is a well-established in vitro model obtained by cultivating mouse embryo. It represents an ideal system to investigate the effect of most prominent epigenetic drugs and agents. Material and methods After isolation, embryos were treated for two hours with 5-azacytidine, Trichostatin A, Valproate, esiNanog, esiOct3/4 and esiTrrap, respectively. The embryos/teratomas were measured at day 0 and for the consequent 7 days of culturing in MEM enriched with rat serum. For analysis of proliferative and apoptotic activity, immunohistochemistry on paraffin embedded embryos/teratomas was performed using anti-Ki-67 and anti-Caspase-3, respectively. Signal intensity was measured by morphometric analysis. For gene expression analyses, embryos/teratomas were pooled into experimental groups from which specific gene related RNA quantity was analysed by both qPCR and ddPCR. Results and discussions Epigenetic modulators reduced significantly embryo/teratoma growth. Most prominent decrease was detected in 5-azacytidine and esiOct3/4 treated embryos/teratomas. Furthermore, 5-azacytidine almost completely disrupted tissue architecture and cellularity. Proliferative activity was not decreased by any epigenetic modulator. EsiNanog, esiTrrap and surprisingly 5-azacytidine, in fact, showed a slight increase in proliferation. Still, 5-azacytidine induced an increase in apoptotic activity as well. Even stronger incitement of apoptosis was found in Valproate treated embryos/teratomas, while other modulators had no effect on apoptotic activity at all. Expression of analysed stemness and differentiation genes panel was significantly disrupted by 5-azacytidine, Valproate and esiOct ¾. Other modulators induced slighter decrease or no change in gene expression. Conclusion This research presents a strong adverse influence of epigenetic modulators on experimental germ cell tumour development. It seems that this effect is consequent to induced change in stemness and differentiation genes expression.

Impact of 5-azacytidine on rat decidual cell proliferation

The DNA demethylating agent 5‐azacytidine (5‐azaC) has a teratogenic influence during rat development influencing both the embryo and the placenta. Our aim was to investigate its impact on early decidual cell proliferation before the formation of placenta. Thus, female Fischer rats received 5‐azaC (5 mg/kg, i.p.) on the 2nd, 5th or 8th day of gestation and the decidual tissues were harvested on gestation day 9. They were then analysed immunohistochemically for expression of cell proliferation marker proliferating cell nuclear antigen (PCNA) in decidual cells and for global DNA methylation using the coupled restriction enzyme digestion, random amplification and pyrosequencing assays. We found that 5‐azaC administered on the 5th and 8th (but not on 2nd) day of gestation led to increased PCNA expression in decidual cells compared with untreated controls. No significant changes in DNA methylation were detected, with either method, in any of the treated rat groups compared with untreated controls. Thus, we conclude that 5‐azaC can stimulate decidual cell proliferation without simultaneously changing global DNA methylation level in treated cells.