Sequence Effect on the Activity of DNAzyme with Covalently Attached Hemin and Their Potential Bioanalytical Application

In this work we investigated the effect of a DNA oligonucleotide sequence on the activity of a DNAzyme with covalently attached hemin. For this purpose, we synthesized seven DNA-hemin conjugates. All DNA-hemin conjugates as well as DNA/hemin complexes were characterized using circular dichroism, determination of melting temperatures and pKa of hemin. We observed that hemin conjugation in most cases led to the formation of parallel G-quadruplexes in the presence of potassium and increased thermal stability of all studied systems. Although the activity of DNA-hemin conjugates depended on the sequence used, the highest activity was observed for the DNA-hemin conjugate based on a human telomeric sequence. We used this DNAzyme for development of “sandwich” assay for detection of DNA sequence. For this assay, we used electric chip which could conduct electricity after silver deposition catalyzed by DNAzyme. This method was proved to be selective towards DNA oligonucleotides with mismatches and could be used for the detection of the target. To prove the versatility of our DNAzyme probe we also performed experiments with streptavidin-coated microplates. Our research proved that DNAzyme with covalently attached hemin can be used successfully in the development of heterogeneous assays.

The Effects of Intensive Neurorehabilitation on Sequence Effect in Parkinson's Disease Patients With and Without Freezing of Gait

Background: The sequence effect (SE), defined as a reduction in amplitude of repetitive movements, is a common clinical feature of Parkinson's disease (PD) and is supposed to be a major contributor to freezing of gait (FOG). During walking, SE manifests as a step-by-step reduction in step length when approaching a turning point or gait destination, resulting in the so-called destination sequence effect (dSE). Previous studies explored the therapeutic effects of several strategies on SE, but none of them evaluated the role of an intensive rehabilitative program.Objectives: Here we aim to study the effects of a 4-week rehabilitative program on dSE in patients with PD with and without FOG.Methods: Forty-three patients (30 males, 70.6 ± 7.5 years old) with idiopathic PD were enrolled. The subjects were divided into two groups: patients with (PD + FOG, n = 23) and without FOG (PD – FOG, n = 20). All patients underwent a standardized 4-week intensive rehabilitation in-hospital program. At hospital admission (T0) and discharge (T1), all subjects were evaluated with an inertial gait analysis for dSE recording.Results: At T0, the dSE was more negative in the PD + FOG group (−0.80 ± 0.6) when compared to the PD – FOG group (−0.39 ± 0.3) (p = 0.007), even when controlling for several clinical and demographic features. At T1, the dSE was reduced in the overall study population (p = 0.001), with a more pronounced improvement in the PD + FOG group (T0: −0.80 ± 0.6; T1: −0.23 ± 0.4) when compared to the PD – FOG group (T0: −0.39 ± 0.3; T1: −0.22 ± 0.5) (p = 0.012). At T1, we described in the overall study population an improvement in speed, cadence, stride duration, and stride length (p = 0.001 for all variables).Conclusions: dSE is a core feature of PD gait dysfunction, specifically in patients with FOG. A 4-week intensive rehabilitative program improved dSE in PD patients, exerting a more notable beneficial effect in the PD + FOG group.

Conflict adaptation and related neuronal processing in Parkinson’s disease

Non-motor symptoms like cognitive impairment are a huge burden for patients with Parkinson’s disease. We examined conflict adaptation by using the congruency sequence effect as an index of adaptation in 17 patients with Parkinson’s disease and 18 healthy controls with an Eriksen flanker task using functional magnet resonance imaging to reveal possible differences in executive function performance. We observed overall increased response times in patients with Parkinson’s disease compared to healthy controls. A flanker interference effect and congruency sequence effect occurred in both groups. A significant interaction of current and previous trial type was revealed, but no effect of response sequence concerning left or right motor responses. Therefore, top-down conflict monitoring processes are likely the main contributors leading to the congruency sequence effect in our paradigm. In both groups incongruent flanker events elicited activation in the middle temporal gyrus, inferior parietal cortex, dorsolateral prefrontal cortex and the insula in contrast to congruent flanker events. A psychophysiological interactions analysis revealed increased functional connectivity of inferior parietal cortex as a seed to the left prefrontal thalamus during incongruent vs. congruent and neutral stimuli in patients with Parkinson’s disease that may reflect compensatory facilitating action selection processes. We conclude that patients with Parkinson’s disease exhibit conflict adaptation comparable to healthy controls when investigated while receiving their usual medication.