Purpose: The primary aim of this study was to identify specific miRNAs with aberrant expression in hippocampal tissue of mesial temporal lobe epilepsy (mTLE) patients with hippocampal sclerosis (HS). Method: We analyzed miRNA isolated from paraffin embedded tissue slices of hippocampal complex from 22 mTLE +HS patients (14 left-sided, 8 right-sided) and 9 post mortem controls. To characterize the miRNAs highly prone to be involved in the pathological changes in the mTLE+HS brain tissue we performed the expression analysis combining the next generation sequencing (NGS) and novel specific technique for miRNA quantification miQPCR. Results: By both methodical approaches we confirmed significant upregulation of miR-129-1-3p, 129-2-3p, 142-3p, 191-5p, 193b-3p, 195-5p, 374b-5p, 451a, and 490-3p in mTLE+HS patients. The levels of miRNA-184 were significantly higher in control subjects. Differential expression levels in mTLE+ HS patients were identified also in the case of 10 miRNA species never considered to play a role in epilepsy before. Conclusion: Analytical approach combining NGS with miQPCR used for the first time for analysis of human epileptic foci led to reliable identification of miRNA with altered expression in mTLE +HS patients.
Throughout the twenty-first century, the view on inclusion bodies (IBs) has shifted from undesired by-products towards a targeted production strategy for recombinant proteins. Inclusion bodies can easily be separated from the crude extract after cell lysis and contain the product in high purity. However, additional solubilization and refolding steps are required in the processing of IBs to recover the native protein. These unit operations remain a highly empirical field of research in which processes are developed on a case-by-case basis using elaborate screening strategies. It has been shown that a reduction in denaturant concentration during protein solubilization can increase the subsequent refolding yield due to the preservation of correctly folded protein structures. Therefore, many novel solubilization techniques have been developed in the pursuit of mild solubilization conditions that avoid total protein denaturation. In this respect, ionic liquids have been investigated as promising agents, being able to solubilize amyloid-like aggregates and stabilize correctly folded protein structures at the same time. This review briefly summarizes the state-of-the-art of mild solubilization of IBs and highlights some challenges that prevent these novel techniques from being yet adopted in industry. We suggest mechanistic models based on the thermodynamics of protein unfolding with the aid of molecular dynamics simulations as a possible approach to solve these challenges in the future.
Objective In the present study, we aimed to investigate depth electroencephalographic (EEG) recordings in a large cohort of patients with drug‐resistant epilepsy and to focus on interictal very high‐frequency oscillations (VHFOs) between 500Hz and 2kHz. We hypothesized that interictal VHFOs are more specific biomarkers for epileptogenic zone compared to traditional HFOs. Methods Forty patients with focal epilepsy who underwent presurgical stereo‐EEG (SEEG) were included in the study. SEEG data were recorded with a sampling rate of 25kHz, and a 30‐minute resting period was analyzed for each patient. Ten patients met selected criteria for analyses of correlations with surgical outcome: detection of interictal ripples (Rs), fast ripples (FRs), and VHFOs; resective surgery; and at least 1 year of postoperative follow‐up. Using power envelope computation and visual inspection of power distribution matrixes, electrode contacts with HFOs and VHFOs were detected and analyzed. Results Interictal very fast ripples (VFRs; 500–1,000Hz) were detected in 23 of 40 patients and ultrafast ripples (UFRs; 1,000–2,000Hz) in almost half of investigated subjects (n = 19). VFRs and UFRs were observed only in patients with temporal lobe epilepsy and were recorded exclusively from mesiotemporal structures. The UFRs were more spatially restricted in the brain than lower‐frequency HFOs. When compared to R oscillations, significantly better outcomes were observed in patients with a higher percentage of removed contacts containing FRs, VFRs, and UFRs. Interpretation Interictal VHFOs are relatively frequent abnormal phenomena in patients with epilepsy, and appear to be more specific biomarkers for epileptogenic zone when compared to traditional HFOs. Ann Neurol 2017;82:299–310
Meduloblastom a multiformni glioblastom představuji zcela odlisne typy mozkových tumorů v parametrech věku pacientů a lokalizace
výskytu. Sekundarni glioblastom představuje nejcastějsi radioterapii indukovaný solidni tumor. Prognoza těchto sekundarnich malignit
je navzdory agresivni multimodalni terapii stale velmi nepřizniva. Autorka prezentuje kazuistiku sestnactileteho chlapce, u ktereho vznikl
sekundarni glioblastom zadni jamy lebni za 6 let po kombinovane lecbě pro meduloblastom v teže lokalizaci. Uvadi klinicke projevy,
patologicke a cytogeneticke nalezy a lecebne možnosti.
Likvorea se u děti vyskytuje převažně u těžkých
kraniocerebralnich poraněni při silnem narazu do překažky nebo
po padu těžkeho předmětu. Tyto urazy jsou casto doprovazeny
mnohocetnými frakturami lbi a oblicejoveho skeletu. U dětských
pacientů je zaznamenan meně castý výskyt likvorey než u
dospělých, což zdůvodňujeme vyssi pružnosti lebky a
nedokoncenou pneumatizaci vedlejsich nosnich dutin u malých
děti. Podle nasich zkusenosti při konzervativni lecbě likvorea
zpravidla vymizi do 1 týdne od urazu. Přesto likvorea
představuje zavažný pourazový stav, který v důsledku poruseni
bariery mezi centralnim nervovým systemem a zevnim prostředim
pacienta ohrožuje intrakranialni infekci.
In protein refolding processes the scarce availability of online measurements hampers effective process monitoring. In this work we developed a mechanistic soft-sensor for protein refolding based on online intrinsic fluorescence measurements of tryptophan and tyrosine. In validation experiments using two model proteins, lactate dehydrogenase (LDH) and galactose oxidase, the soft-sensor showed accurate estimates for the prediction of the total sum of folding products (NRMSE <6.1%) by calculating the changing rate of the average emission wavelength. For refolding of the enzyme LDH it was possible to obtain separate predictions of native protein and insoluble aggregates. The soft-sensor design was further extended by a model-based observer approach using particle filtering to incorporate kinetic formulations as well as physical constraints. The novel approach enabled the analysis of kinetic mechanisms during rapid reaction dynamics and can therefore be seen as an enabler to achieve a better understanding of kinetic refolding mechanisms.
