Miracle-dz/UniFAR-FTUs huggingface dataset:
Serum miR‐206 and other muscle‐specific microRNAs as non‐invasive biomarkers for Duchenne muscular dystrophy

Creatine kinase has been utilized as a diagnostic marker for Duchenne muscular dystrophy (DMD), but it correlates less well with the DMD pathological progression. In this study, we hypothesized that muscle‐specific microRNAs (miR‐1, ‐133, and ‐206) in serum may be useful for monitoring the DMD pathological progression, and explored the possibility of these miRNAs as potential non‐invasive biomarkers for the disease. By using real‐time quantitative reverse transcription–polymerase chain reaction in a randomized and controlled trial, we detected that miR‐1, ‐133, and ‐206 were significantly over‐expressed in the serum of 39 children with DMD (up to 3.20 ± 1.20, 2−ΔΔCt): almost 2‐ to 4‐fold enriched in comparison to samples from the healthy controls (less than 1.15 ± 0.34, 2−ΔΔCt). To determine whether these miRNAs were related to the clinical features of children with DMD, we analyzed the associations compared to creatine kinase. There were very good inverse correlations between the levels of these miRNAs, especially miR‐206, and functional performances: high levels corresponded to low muscle strength, muscle function, and quality of life. Moreover, by receiver operating characteristic curves analyses, we revealed that these miRNAs, especially miR‐206, were able to discriminate DMD from controls. Thus, miR‐206 and other muscle‐specific miRNAs in serum are useful for monitoring the DMD pathological progression, and hence as potential non‐invasive biomarkers for the disease.
Häufigkeit von Doppeluntersuchungen in der B-Bild-sonographischen Abteilung einer radiologischen Universitätsklinik

PURPOSE What is the percentage of repeat examinations in the ultrasonographic section of a radiological university department? METHODS All 4116 patients, who were first examined in the sonographic section of the Radiologic Department of the University of Cologne from 5/97 to 5/98, were asked by the medical staff, whether, how often and by whom ultrasonographic, CT or MR examinations of the same organ system had been carried out during the last 4 weeks without evidence of clinical changes. The data were documented using a structured questionnaire. RESULTS Sonographic repeated examinations documented in 443 (10.8%) of the 4116 consultations. The proportion of in- to outpatients with repeat examinations was 26.0% to 74.0%. Contrarily, the repeated ultrasonographies were done by practitioners in 62.3% and by the university staff in 33.2% (radiology 48.2%). With regard to the 4116 consultations, the abdomen (81.1%) was the organ system most often examined repeatedly. Computed tomographies were already done or planned in 305 of 4116 patients (7.4%) and MR-tomographies were already done or planned in 57 patients (1.4%). CONCLUSION The rate of repeated ultrasonographic examinations in a radiological university hospital was 19.6%.
Bidirectional interactions between NOX2‐type NADPH oxidase and the F‐actin cytoskeleton in neuronal growth cones

NADPH oxidases are important for neuronal function but detailed subcellular localization studies have not been performed. Here, we provide the first evidence for the presence of functional NADPH oxidase 2 (NOX2)‐type complex in neuronal growth cones and its bidirectional relationship with the actin cytoskeleton. NADPH oxidase inhibition resulted in reduced F‐actin content, retrograde F‐actin flow, and neurite outgrowth. Stimulation of NADPH oxidase via protein kinase C activation increased levels of hydrogen peroxide in the growth cone periphery. The main enzymatic NADPH oxidase subunit NOX2/gp91phox localized to the growth cone plasma membrane and showed little overlap with the regulatory subunit p40phox. p40phox itself exhibited colocalization with filopodial actin bundles. Differential subcellular fractionation revealed preferential association of NOX2/gp91phox and p40phox with the membrane and the cytoskeletal fraction, respectively. When neurite growth was evoked with beads coated with the cell adhesion molecule apCAM, we observed a significant increase in colocalization of p40phox with NOX2/gp91phox at apCAM adhesion sites. Together, these findings suggest a bidirectional functional relationship between NADPH oxidase activity and the actin cytoskeleton in neuronal growth cones, which contributes to the control of neurite outgrowth.
Neuroactive steroid levels in plasma and cerebrospinal fluid of male multiple sclerosis patients

Neuroactive steroid family includes molecules synthesized in peripheral glands (i.e., hormonal steroids) and directly in the nervous system (i.e., neurosteroids) which are key regulators of the nervous function. As already reported in clinical and experimental studies, neurodegenerative diseases affect the levels of neuroactive steroids. However, a careful analysis comparing the levels of these molecules in cerebrospinal fluid (CSF) and in plasma of multiple sclerosis (MS) patients is still missing. To this aim, the levels of neuroactive steroids were evaluated by liquid chromatography‐tandem mass spectrometry in CSF and plasma of male adults affected by Relapsing‐Remitting MS and compared with those collected in control patients. An increase in pregnenolone and isopregnanolone levels associated with a decrease in progesterone metabolites, dihydroprogesterone, and tetrahydroprogesterone was observed in CSF of MS patients. Moreover, an increase of 5α‐androstane‐3α,17β‐diol and of 17β‐estradiol levels associated with a decrease of dihydrotestosterone also occurred. In plasma, an increase in pregnenolone, progesterone, and dihydrotestosterone and a decrease in dihydroprogesterone and tetrahydroprogesterone levels were reported. This study shows for the first time that the levels of several neuroactive steroids, and particularly those of progesterone and testosterone metabolites, are deeply affected in CSF of relapsing‐remitting MS male patients.
Inhibition of PaCaMKII‐E isoform in the dorsal unpaired median neurosecretory cells of cockroach reduces nicotine‐ and clothianidin‐induced currents

Cellular responses to Ca2+ require intermediary proteins such as calcium/calmodulin‐dependent protein kinase II (CaMKII), which transduces the signal into downstream effects. We recently demonstrated that the cockroach genome encodes five different CaMKII isoforms, and only PaCaMKII‐E isoform is specifically expressed in the dorsal unpaired median neurosecretory cells. In the present study, using antisense oligonucleotides, we demonstrated that PaCaMKII‐E isoform inhibition reduced nicotine‐induced currents through α‐bungarotoxin‐sensitive and ‐insensitive nicotinic acetylcholine receptor subtypes. Specifically, PaCaMKII‐E isoform is sufficient to repress nicotinic current amplitudes as a result of its depression by antisense oligonucleotides. Similar results were found using the neonicotinoid insecticide clothianidin, which acted as a full agonist of dorsal unpaired median neuron nicotinic acetylcholine receptors. Clothianidin current amplitudes are strongly reduced under bath application of PaCaMKII‐E antisense oligonucleotides but no significant results are found with α‐bungarotoxin co‐applied, demonstrating that CaMKII‐E isoform affects nicotine currents through α‐bungarotoxin‐sensitive and ‐insensitive receptor subtypes whereas clothianidin currents are reduced via α‐bungarotoxin‐insensitive receptors. In addition, we found that intracellular calcium increase induced by nicotine and clothianidin were reduced by PaCaMKII‐E antisense oligonucleotides, demonstrating that intracellular calcium increase induced by nicotine and clothianidin are affected by PaCaMKII‐E inhibition.
Neurotherapeutic effects of novel HO‐1 inhibitors in vitro and in a transgenic mouse model of Alzheimer's disease

Heme oxygenase‐1 (HO‐1) encoded by the HMOX1 gene is a 32‐kDa stress protein that catabolizes heme to biliverdin, free iron, and carbon monoxide (CO). Glial HO‐1 is over‐expressed in the CNS of subjects with Alzheimer's disease (AD), Parkinson's disease (PD), and multiple sclerosis (MS). The HMOX1 gene is exquisitely sensitive to oxidative stress and is induced in brain and other tissues in various models of disease and trauma. Induction of the glial HMOX1 gene may lead to pathological brain iron deposition, intracellular oxidative damage, and bioenergetic failure in AD and other human CNS disorders such as PD and MS. Therefore, targeted suppression of glial HO‐1 hyperactivity may prove to be a rational and effective therapeutic intervention in AD and related neurodegenerative disorders. In this study, we report the effects of QC‐47, QC‐56, and OB‐28, novel azole‐based competitive and reversible inhibitors of HO‐1, on oxidative damage to whole‐cell and mitochondrial compartments in rat astrocytes transfected with the HMOX1 gene. We also report the effect of OB‐28 on the behavior and neuropathology of APPswe/PS1∆E9 mice. OB‐28 was found to reduce oxidative damage to whole‐cell and mitochondrial compartments in rat astrocytes transfected with the HMOX1 gene. Moreover, OB‐28 was found to significantly counter behavioral deficits and neuropathological alterations in APPswe/PS1∆E9 mice. Attenuation of AD‐associated behavioral deficits and neuropathological changes suggests that HO‐1 may be a promising target for neuroprotective intervention in AD and other neurodegenerative diseases.
Cleaning up after ICH: the role of Nrf2 in modulating microglia function and hematoma clearance

As a consequence of intracerebral hemorrhage (ICH), blood components enter brain parenchyma causing progressive damage to the surrounding brain. Unless hematoma is cleared, the reservoirs of blood continue to inflict injury to neurovascular structures and blunt the brain repair processes. Microglia/macrophages (MMΦ) represent the primary phagocytic system that mediates the cleanup of hematoma. Thus, the efficacy of phagocytic function by MMΦ is an essential step in limiting ICH‐mediated damage. Using primary microglia to model red blood cell (main component of hematoma) clearance, we studied the role of transcription factor nuclear factor‐erythroid 2 p45‐related factor 2 (Nrf2), a master‐regulator of antioxidative defense, in the hematoma clearance process. We showed that in cultured microglia, activators of Nrf2 (i) induce antioxidative defense components, (ii) reduce peroxide formation, (iii) up‐regulate phagocytosis‐mediating scavenger receptor CD36, and (iv) enhance red blood cells (RBC) phagocytosis. Through inhibiting Nrf2 or CD36 in microglia, by DNA decoy or neutralizing antibody, we documented the important role of Nrf2 and CD36 in RBC phagocytosis. Using autologous blood injection ICH model to measure hematoma resolution, we showed that Nrf2 activator, sulforaphane, injected to animals after the onset of ICH, induced CD36 expression in ICH‐affected brain and improved hematoma clearance in rats and wild‐type mice, but expectedly not in Nrf2 knockout (KO) mice. Normal hematoma clearance was impaired in Nrf2‐KO mice. Our experiments suggest that Nrf2 in microglia play an important role in augmenting the antioxidative capacity, phagocytosis, and hematoma clearance after ICH.
Individualized Music

Abstract Purpose: The present study examined the effects of providing cognitively intact nursing home residents with music of their own choosing on immediate satisfaction and global quality of life. Method and Design: The design was a pre- and post-test intervention study. Nursing home residents were presented with music of their own choosing and asked to respond to questions concerning their previous music experience, their enjoyment of the music presentation, and their overall quality of life. Results: Residents reported extensive past music experience and had a diverse array of music p
Susceptibility to excitotoxicity in aged hippocampal cultures and neuroprotection by non‐steroidal anti‐inflammatory drugs: role of mitochondrial calcium

Brain damage after insult and cognitive decline are related to excitotoxicity and strongly influenced by aging, yet mechanisms of aging‐dependent susceptibility to excitotoxicity are poorly known. Several non‐steroidal anti‐inflammatory drugs (NSAIDs) may prevent excitotoxicity and cognitive decline in the elderly by an unknown mechanism. Interestingly, after several weeks in vitro, hippocampal neurons display important hallmarks of neuronal aging in vivo. Accordingly, rat hippocampal neurons cultured for several weeks were used to investigate mechanisms of aging‐related susceptibility to excitotoxicity and neuroprotection by NSAIDs. We found that NMDA increased cytosolic Ca2+ concentration in young, mature and aged neurons but only promoted apoptosis in aged neurons. Resting Ca2+ levels and responses to NMDA increased with time in culture which correlated with changes in expression of NMDA receptor subunits. In addition, NMDA promoted mitochondrial Ca2+ uptake only in aged cultures. Consistently, specific inhibition of mitochondrial Ca2+ uptake decreased apoptosis. Finally, we found that a series of NSAIDs depolarized mitochondria and inhibited mitochondrial Ca2+ overload, thus preventing NMDA‐induced apoptosis in aged cultures. We conclude that mitochondrial Ca2+ uptake is critical for age‐related susceptibility to excitotoxicity and neuroprotection by NSAIDs.
Molecular mechanisms of non‐transferrin‐bound and transferring‐bound iron uptake in primary hippocampal neurons

The molecular mechanisms of iron trafficking in neurons have not been elucidated. In this study, we characterized the expression and localization of ferrous iron transporters Zip8, Zip14 and divalent metal transporter 1 (DMT1), and ferrireductases Steap2 and stromal cell‐derived receptor 2 in primary rat hippocampal neurons. Steap2 and Zip8 partially co‐localize, indicating these two proteins may function in Fe3+ reduction prior to Fe2+ permeation. Zip8, DMT1, and Steap2 co‐localize with the transferrin receptor/transferrin complex, suggesting they may be involved in transferrin receptor/transferrin‐mediated iron assimilation. In brain interstitial fluid, transferring‐bound iron (TBI) and non‐transferrin‐bound iron (NTBI) exist as potential iron sources. Primary hippocampal neurons exhibit significant iron uptake from TBI (Transferrin‐59Fe3+) and NTBI, whether presented as 59Fe2+‐citrate or 59Fe3+‐citrate; reductase‐independent 59Fe2+ uptake was the most efficient uptake pathway of the three. Kinetic analysis of Zn2+ inhibition of Fe2+ uptake indicated that DMT1 plays only a minor role in the uptake of NTBI. In contrast, localization and knockdown data indicate that Zip8 makes a major contribution. Data suggest also that cell accumulation of 59Fe from TBI relies at least in part on an endocytosis‐independent pathway. These data suggest that Zip8 and Steap2 play a major role in iron accumulation from NTBI and TBI by hippocampal neurons. Analysis of the expression and localization of known iron uptake transporters demonstrated that Zip8 makes a major contribution to iron accumulation in primary cultures of rat embryonic hippocampal neurons. These cells exhibit uptake pathways for ferrous and ferric iron (non‐transferrin‐bound iron, NTBI in figure) and for transferrin‐bound iron; the ferrireductases Steap2 and SDR2 support the uptake of ferric iron substrates. Zip8 and Steap2 are strongly expressed in the plasma membrane of both soma and processes, implying a crucial role in iron accumulation from NTBI and transferrin‐bound iron (TBI) by hippocampal neurons.
Noise‐induced cochlear F‐actin depolymerization is mediated via ROCK2/p‐ERM signaling

Our previous work has suggested that traumatic noise activates Rho‐GTPase pathways in cochlear outer hair cells (OHCs), resulting in cell death and noise‐induced hearing loss (NIHL). In this study, we investigated Rho effectors, Rho‐associated kinases (ROCKs), and the targets of ROCKs, the ezrin‐radixin‐moesin (ERM) proteins, in the regulation of the cochlear actin cytoskeleton using adult CBA/J mice under conditions of noise‐induced temporary threshold shift (TTS) and permanent threshold shift (PTS) hearing loss, which result in changes to the F/G‐actin ratio. The levels of cochlear ROCK2 and p‐ERM decreased 1 h after either TTS‐ or PTS‐noise exposure. In contrast, ROCK2 and p‐ERM in OHCs decreased only after PTS‐, not after TTS‐noise exposure. Treatment with lysophosphatidic acid, an activator of the Rho pathway, resulted in significant reversal of the F/G‐actin ratio changes caused by noise exposure and attenuated OHC death and NIHL. Conversely, the down‐regulation of ROCK2 by pretreatment with ROCK2 siRNA reduced the expression of ROCK2 and p‐ERM in OHCs, exacerbated TTS to PTS, and worsened OHC loss. Additionally, pretreatment with siRNA against radixin, an ERM protein, aggravated TTS to PTS. Our results indicate that a ROCK2‐mediated ERM‐phosphorylation signaling cascade modulates noise‐induced hair cell loss and NIHL by targeting the cytoskeleton.
Functional characterization of mutants in the transmembrane domains of the rat P2X7 receptor that regulate pore conductivity and agonist sensitivity

In the sustained presence of agonist, the opening of P2X7R channel is followed by pore dilatation, which causes an increase in its permeability to larger organic cations, accompanied by receptor sensitization. To explore the molecular mechanisms by which the conductivity and sensitivity are increased, we analyzed the electrophysiological properties and YO‐PRO‐1 uptake of selected alanine mutants in the first and second transmembrane domains of the rat P2X7R. Substitution of residues Y40, F43, G338, and D352 with alanine reduced membrane trafficking, and the D352A was practically non‐functional. The Y40A and F43A mutants that were expressed in the membrane lacked pore dilation ability. Moreover, the Y40A and Y40F displayed desensitization, whereas the Y40W partially recovered receptor function. The G338A/S mutations favored the open state of the channel and displayed instantaneous permeability to larger organic cations. The G338P was non‐functional. The L341A and G345A displayed normal trafficking, current amplitude, and sensitization, but both mutations resulted in a decreased pore formation and dye uptake. These results showed that the increase in P2X7R conductivity and sensitivity is critically dependent on residues Y40 and F43 in the TM1 domain and that the region located at the intersection of TM2 helices controls the rate of large pore opening. We investigated the mechanism of the proapoptotic receptor P2X7R's large pore opening and its sensitization. We found that aromatic residues in the upper part of the first transmembrane domain (TM1) are critical for both the P2X7R channel pore opening and receptor sensitization, and residues located at or below the intersection of the second transmembrane domains (TM2) control the rate of pore opening. These findings identify new residues involved in pore formation of P2X7R.
Functional morphology of digestion in the stomachless, piscivorous needlefishes Tylosurus gavialoides and Strongylura leiura ferox (Teleostei: Beloniformes)

Belonidae are unusual in that they are carnivorous but lack a stomach and have a straight, short gut. To develop a functional morphological model for this unusual system the gut contents and alimentary tract morphology of Tylosurus gavialoides and Strongylura leiura ferox were investigated. The posterior orientation of the majority of the pharyngeal teeth supports the swallowing of whole large prey, but not their mastication. Mucogenic cells are abundant in the mucosa lining, particularly the esophagus, and their secretions are likely to protect the gut lining from damage while lubricating passage of the prey. Esophagus, anterior intestine, posterior intestine, and rectum all have highly reticulate mucosae. The anterior three gut sections are distensible to accommodate the passage of prey. However, following ingestion large prey are passed to the highly distensible posterior intestine where they rest head first against the ileorectal valve. Alimentary pH ranges from neutral to weakly acidic. Fish prey is digested head first with the head being largely digested while the remainder of the body is still intact. The nondistensibility of the rectum and the small aperture provided by the ileorectal valve suggest the products of intestinal digestion are either small particulates or fluids that pass into rectum where they are absorbed. J. Morphol. 2009. © 2009 Wiley‐Liss, Inc.
Drebrin depletion alters neurotransmitter receptor levels in protein complexes, dendritic spine morphogenesis and memory‐related synaptic plasticity in the mouse hippocampus

Drebrin an actin‐bundling key regulator of dendritic spine genesis and morphology, has been recently proposed as a regulator of hippocampal glutamatergic activity which is critical for memory formation and maintenance. Here, we examined the effects of genetic deletion of drebrin on dendritic spine and on the level of complexes containing major brain receptors. To this end, homozygous and heterozygous drebrin knockout mice generated in our laboratory and related wild‐type control animals were studied. Level of protein complexes containing dopamine receptor D1/dopamine receptor D2, 5‐hydroxytryptamine receptor 1A (5‐HT1AR), and 5‐hydroxytryptamine receptor 7 (5‐HT7R) were significantly reduced in hippocampus of drebrin knockout mice whereas no significant changes were detected for GluR1, 2, and 3 and NR1 as examined by native gel‐based immunoblotting. Drebrin depletion also altered dendritic spine formation, morphology, and reduced levels of dopamine receptor D1 in dendritic spines as evaluated using immunohistochemistry/confocal microscopy. Electrophysiological studies further showed significant reduction in memory‐related hippocampal synaptic plasticity upon drebrin depletion. These findings provide unprecedented experimental support for a role of drebrin in the regulation of memory‐related synaptic plasticity and neurotransmitter receptor signaling, offer relevant information regarding the interpretation of previous studies and help in the design of future studies on dendritic spines. We examined effect of genetic deletion of drebrin, which an actin‐bundling key regulator of dendritic spine genesis and morphology, on dendritic spine density, maturity, level of complexes containing major brain receptors and also, in synaptic plasticity. These findings support for a role of drebrin in the regulation of memory‐related synaptic plasticity and neurotransmitter receptors signaling in dendritic spines.
N‐acetyl‐l‐tryptophan, but not N‐acetyl‐d‐tryptophan, rescues neuronal cell death in models of amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by progressive motor neuron loss. Evidence suggests that mitochondrial dysfunction, apoptosis, oxidative stress, inflammation, glutamate excitotoxicity, and proteasomal dysfunction are all responsible for ALS pathogenesis. N‐acetyl‐tryptophan has been identified as an inhibitor of mitochondrial cytochrome c release and therefore is a potential neuroprotective agent. By quantifying cell death, we demonstrate that N‐acetyl‐l‐tryptophan (L‐NAT) and N‐acetyl‐DL‐tryptophan are neuroprotective in NSC‐34 motor neuron‐like cells and/or primary motor neurons, while their isomer N‐acetyl‐d‐tryptophan has no protective effect. These findings are consistent with energy minimization and molecular modeling analysis, confirming that L‐NAT generates the most stable complex with the neurokinin‐1 receptor (NK‐1R). L‐NAT inhibits the secretion of Substance P and IL‐1β (Enzyme‐Linked Immunosorbent Assay and/or dot blots) and mitochondrial dysfunction by effectively inhibiting the release of cytochrome c/Smac/AIF from mitochondria into the cytoplasm and activation of apoptotic pathways, including the activation of caspase‐1, ‐9, and ‐3, as well as proteasomal dysfunction through restoring chymotrypsin‐like, trypsin‐like, and caspase‐like proteasome activity. These data provide insight into the molecular mechanisms by which L‐NAT offers neuroprotection in models of ALS and suggest its potential as a novel therapeutic strategy for ALS.