Journal: Physiological Research

Article Title: The Severity of Muscle Performance Deterioration in Sarcopenia Correlates With Circulating Muscle Tissue-Specific miRNAs

doi: 10.33549/physiolres.934778

Figure Lengend Snippet: List of microRNA primers (Qiagen GmbH, Germany)

Article Snippet: The PCR reaction was performed in a StepOnePlus Real-Time PCR System (Life Technologies, USA). lists the PCR primers used in this study. table ft1 table-wrap mode="anchored" t5 caption a7 microRNA LNA PCR primer ID microRNA LNA PCR primer ID cel-miR-39-3p YP00203952 hsa-miR-133a-3p YP00204788 hsa-miR-1-3p YP00204344 hsa-miR-133b YP00206058 hsa-miR-206 YP00206073 hsa-miR-208b-3p YP00204636 hsa-miR-29a-3p YP00204698 hsa-miR-499-5p YP00205935 hsa-miR-29b-3p YP00204679 Open in a separate window List of microRNA primers (Qiagen GmbH, Germany) Statistical analysis Statistical evaluation was performed by Graphpad using one way ANOVA test and Chi test.

Techniques:

Journal: Neuropathology and Applied Neurobiology

Article Title: Chronic activation of anti‐oxidant pathways and iron accumulation in epileptogenic malformations

doi: 10.1111/nan.12596

Figure Lengend Snippet: Higher expression of miR155 and HO‐1 in TSC and FCD IIb. ( A ) Expression of miR155 was predominately found in neurones of the grey matter (GM) with low expression in glia of the white matter (WM) (arrowheads). ( B, C ) In contrast, miR155 expression was predominantly found in GFAP‐positive cells with glial morphology and giant/balloon cells in FCD IIb and TSC as compared to control, whereas expression in dysmorphic neurones did not differ from control. ( D ) Expression of total miR155 in TSC tissue was higher than in control. ( E, F ) Moreover, HO‐1 RNA expression was higher in surgically resected tuber tissue from TSC patients compared to autoptic control, while HO‐1 protein expression was high only in a subset of patients. ( G, H ) miR155 expression was not different between autopsy control tissue and FCD IIb, while HO‐1 RNA was higher. Mann–Whitney U test. Data are expressed relative to expression observed in controls. Error bars represent range; ** P < 0.01. n = 8 (Autopsy control, FCD IIb), n = 10 (TSC). Scale bar 100 µm in A (representative for A‐C), arrows = dysmorphic neurones, arrowheads = glia, asterisk = balloon/giant cells. [Colour figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: For transfection, cultures were transfected with either mimic negative control (Figure ), miR155 mimic (Applied Biosystems, Carlsbad, CA, USA) or antisense miR155 LNA oligonucleotide (miR155 antagomiR, Ribotask ApS, Odense, Denmark) (Table ).

Techniques: Expressing, Control, RNA Expression, MANN-WHITNEY

Journal: Neuropathology and Applied Neurobiology

Article Title: Chronic activation of anti‐oxidant pathways and iron accumulation in epileptogenic malformations

doi: 10.1111/nan.12596

Figure Lengend Snippet: Higher expression of 4‐HNE, HO‐1 and miR155 in Tsc1 GFAP−/− mice precedes the development of seizures. ( A, B ) Expression of 4‐HNE in the hippocampus (HC) and cortex (Cx) of 2‐week‐old control mice was low while it was detectable in 2‐week‐old Tsc1 GFAP−/− mice before seizure onset, however, only in the hippocampus mainly perivascular and in GFAP and occasionally NeuN expressing cells, but not Iba‐1‐positive cells ( B 1 –B 3 ). ( C, D ) 4‐HNE expression was higher in the hippocampus of 2‐month‐old control mice after seizure onset compared to 2‐week‐old control mice before seizure onset. Two‐month‐old Tsc1 GFAP−/− mice with recurrent seizures displayed high 4‐HNE expression in the hippocampus and cortex with similar perivascular expression and co‐localization with GFAP‐positive cells as in 2‐week‐old mice ( D 1 –D 3 ). ( E, G ) HO‐1 was moderately expressed in the hippocampus and cortex of 2‐week‐old and 2‐month‐old control mice. ( F ) In contrast, 2‐week‐old Tsc1 GFAP−/− mice showed very high expression in sparsely distributed GFAP‐positive cells mainly in the hippocampus before seizure onset (arrowheads, F 1 ). Iba‐1‐positive cells were in close proximity to the HO‐1 expressing cells, but did not show co‐localization ( F 2,3 ). ( H ) In 2‐month‐old Tsc1 GFAP−/− mice with recurrent seizures the number of cells displaying strong HO‐1 expression was higher than in animals before seizure onset and could also be found in the cortex. ( I–J ) Quantification of 4‐HNE OD revealed higher 4‐HNE reactivity in 2‐week‐old animals in the HC and in HC and Cx in 2‐month‐old mice. ( K ) Additionally, the count of cells with strong HO‐1 expression increased in the HC and Cx in 2‐month‐old mice after seizure development ( F 1 , H 1 ). ( L–O ) RNA quantification of miR155 and HO‐1 in the hippocampus and cortex revealed higher expression in Tsc1 GFAP−/− mice already before seizure onset compared to control, which was even higher in mice after the development of recurrent seizures. Scale bars: 500 µm (hippocampus) and 100 µm (cortex) in A, 50µm in B 1 (representative of B 1 –B 3 , D 1 –D 3 ) and 20 µm in insert in F 1 (representative of F 1 –F 3 , H 1 –H 3 ). Mann–Whitney U test. Data are expressed relative to expression observed in WT for the respective age group and presented as individual data points as well as in box plots. Error bars represent range; * P < 0.05, ** P < 0.01 (L‐O ## P < 0.01 2‐week‐old vs. 2‐month‐old Tsc1 GFAP−/− mice). n = 5 animals per group. [Colour figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: For transfection, cultures were transfected with either mimic negative control (Figure ), miR155 mimic (Applied Biosystems, Carlsbad, CA, USA) or antisense miR155 LNA oligonucleotide (miR155 antagomiR, Ribotask ApS, Odense, Denmark) (Table ).

Techniques: Expressing, Control, MANN-WHITNEY

Journal: Neuropathology and Applied Neurobiology

Article Title: Chronic activation of anti‐oxidant pathways and iron accumulation in epileptogenic malformations

doi: 10.1111/nan.12596

Figure Lengend Snippet: In vitro , human foetal astrocytes displayed different response to acute vs. chronic OS and chronic HO‐1 expression can induce genes involved in iron regulation. ( A ) Acute OS rapidly induced gene expression of Nrf‐2 targets HO‐1 and xCT, while NF‐κB signalling genes TLR‐4 and TAB‐2 were downregulated. Chronic OS increased expression of HO‐1 and xCT. However, the HO‐1 increase was lower than after acute OS. Moreover, in contrast to acute OS TLR‐4, TAB‐2 and MYD88 expression was increased. ( B ) Exposure to acute OS induced rapid expression of γH2A.X in the nucleus of human foetal astrocytes compared to control (B 2 ). Quantification of cells displaying >10 γH2A.X foci could not be detected in control cells, while acute OS induced expression in approximately 55% of cells (B 3 ). ( C, D, H, I ) Transfection of foetal astrocytes with miR155 mimic induced expression of HO‐1 and xCT even in the absence of OS. This effect was preserved in cells derived from TSC patients and could be reversed by the inhibitor of miR155. ( E ) Additionally, Bach‐1 expression was lower. These effects were independent of the presence of acute (3 h) or chronic (72 h) OS. ( F, G ) Transfection of foetal astrocytes with miR155 mimic for 24 h followed by acute OS had no effect on expression of FTH‐1 and FPN‐1. In contrast, prolonged exposure to the miR155 mimic for 72 h coupled to chronic OS induced FTH‐1 and FPN‐1. ( J ) Moreover, foetal astrocytes exposed to miR155 mimic for 72 h displayed increased susceptibility to high H 2 O 2 concentrations compared to control and miR155 inhibitor transfected cells as measured using the MTT assay. ( K ) Foetal astrocytes stimulated with H 2 O 2 for different time points displayed rapid expression of HO‐1 protein which peaked at 6 h and decreased again after 24 h. In parallel, ferritin expression increased with a delay and remained high even after 24 h. Scale bar is 10 µm in B. Mann–Whitney U test in A, J. Kruskal–Wallis test followed by Dunn’s in C–I, K. Data are expressed relative to expression observed in control groups and the mean value as well as the individual data points are shown. Error bars represent SEM; * P < 0.05, ** P < 0.01, n = 3 independent cultures in duplicates ( n = 379 cells (control) and 442 cells (acute OS) in B 3 ; three single cultures for H, I, K; three independent cultures in quadruplicates for J) per experiment. [Colour figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: For transfection, cultures were transfected with either mimic negative control (Figure ), miR155 mimic (Applied Biosystems, Carlsbad, CA, USA) or antisense miR155 LNA oligonucleotide (miR155 antagomiR, Ribotask ApS, Odense, Denmark) (Table ).

Techniques: In Vitro, Expressing, Gene Expression, Control, Transfection, Derivative Assay, MTT Assay, MANN-WHITNEY

Journal: Current Research in Pharmacology and Drug Discovery

Article Title: Scaling approaches for the prediction of human clearance of LNA-i-mir-221: A retrospective validation

doi: 10.1016/j.crphar.2024.100197

Figure Lengend Snippet: Prediction corrected visual predictive check vs. time after last dose for the final model for LNA-i-miR-221. Prediction intervals for each percentile are estimated across all simulated data and displayed as colored areas (pink for the 50th percentile, blue for the 10th and 90th percentiles). Prediction intervals are computed with a level of 90%. Empirical percentiles: percentiles of the observed data, calculated each unique value of time (bins, bin intervals are defined by vertical violet lines). Outliers are highlighted with red dots and areas. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: LNA-i-miR-221 concentrations were determined in human plasma and urines using validated mass spectrometry analytical methods( ) at Aptuit (with a Lower Limit Of Quantification (LLOQ of 50 ng/mL).

Techniques:

Journal: Current Research in Pharmacology and Drug Discovery

Article Title: Scaling approaches for the prediction of human clearance of LNA-i-mir-221: A retrospective validation

doi: 10.1016/j.crphar.2024.100197

Figure Lengend Snippet: Correlation between predicted (3 compartments – lin CL model) and measured individual concentrations of LNA-i-miR-221 in humans.

Article Snippet: LNA-i-miR-221 concentrations were determined in human plasma and urines using validated mass spectrometry analytical methods( ) at Aptuit (with a Lower Limit Of Quantification (LLOQ of 50 ng/mL).

Techniques: