Insulin signalling regulates Pink1 mRNA localization via modulation of AMPK activity to support PINK1 function in neurons.

Mitochondrial quality control failure is frequently observed in neurodegenerative diseases. The detection of damaged mitochondria by stabilization of PTEN-induced kinase 1 (PINK1) requires transport of Pink1 messenger RNA (mRNA) by tethering it to the mitochondrial surface. Here, we report that inhibition of AMP-activated protein kinase (AMPK) by activation of the insulin signalling cascade prevents Pink1 mRNA binding to mitochondria. Mechanistically, AMPK phosphorylates the RNA anchor complex subunit SYNJ2BP within its PDZ domain, a phosphorylation site that is necessary for its interaction with the RNA-binding protein SYNJ2. Notably, loss of mitochondrial Pink1 mRNA association upon insulin addition is required for PINK1 protein activation and its function as a ubiquitin kinase in the mitophagy pathway, thus placing PINK1 function under metabolic control. Induction of insulin resistance in vitro by the key genetic Alzheimer risk factor apolipoprotein E4 retains Pink1 mRNA at the mitochondria and prevents proper PINK1 activity, especially in neurites. Our results thus identify a metabolic switch controlling Pink1 mRNA localization and PINK1 activity via insulin and AMPK signalling in neurons and propose a mechanistic connection between insulin resistance and mitochondrial dysfunction.

Live-Cell Imaging of RNA Transport in Axons of Cultured Primary Neurons.

The use of fluorescent proteins has revolutionized the study of protein localization and transport. However, the visualization of other molecules and specifically RNA during live-cell imaging remains challenging. In this chapter, we provide guidance to the available methods, their advantages and drawbacks as well as provide a detailed protocol for the detection of RNA transport using the MS2/PP7-split-Venus system for background-free RNA imaging.

Neuronal mitochondria transport Pink1 mRNA via synaptojanin 2 to support local mitophagy.

PTEN-induced kinase 1 (PINK1) is a short-lived protein required for the removal of damaged mitochondria through Parkin translocation and mitophagy. Because the short half-life of PINK1 limits its ability to be trafficked into neurites, local translation is required for this mitophagy pathway to be active far from the soma. The Pink1 transcript is associated and cotransported with neuronal mitochondria. In concert with translation, the mitochondrial outer membrane proteins synaptojanin 2 binding protein (SYNJ2BP) and synaptojanin 2 (SYNJ2) are required for tethering Pink1 mRNA to mitochondria via an RNA-binding domain in SYNJ2. This neuron-specific adaptation for the local translation of PINK1 provides distal mitochondria with a continuous supply of PINK1 for the activation of mitophagy.

Mitochondrial Phenotypes in Parkinson's Diseases-A Focus on Human iPSC-Derived Dopaminergic Neurons.

Established disease models have helped unravel the mechanistic underpinnings of pathological phenotypes in Parkinson's disease (PD), the second most common neurodegenerative disorder. However, these discoveries have been limited to relatively simple cellular systems and animal models, which typically manifest with incomplete or imperfect recapitulation of disease phenotypes. The advent of induced pluripotent stem cells (iPSCs) has provided a powerful scientific tool for investigating the underlying molecular mechanisms of both familial and sporadic PD within disease-relevant cell types and patient-specific genetic backgrounds. Overwhelming evidence supports mitochondrial dysfunction as a central feature in PD pathophysiology, and iPSC-based neuronal models have expanded our understanding of mitochondrial dynamics in the development and progression of this devastating disorder. The present review provides a comprehensive assessment of mitochondrial phenotypes reported in iPSC-derived neurons generated from PD patients' somatic cells, with an emphasis on the role of mitochondrial respiration, morphology, and trafficking, as well as mitophagy and calcium handling in health and disease. Furthermore, we summarize the distinguishing characteristics of vulnerable midbrain dopaminergic neurons in PD and report the unique advantages and challenges of iPSC disease modeling at present, and for future mechanistic and therapeutic applications.

Kill one or kill the many: interplay between mitophagy and apoptosis.

Mitochondria are key players of cellular metabolism, Ca2+ homeostasis, and apoptosis. The functionality of mitochondria is tightly regulated, and dysfunctional mitochondria are removed via mitophagy, a specialized form of autophagy that is compromised in hereditary forms of Parkinson's disease. Through mitophagy, cells are able to cope with mitochondrial stress until the damage becomes too great, which leads to the activation of pro-apoptotic BCL-2 family proteins located on the outer mitochondrial membrane. Active pro-apoptotic BCL-2 proteins facilitate the release of cytochrome c from the mitochondrial intermembrane space (IMS) into the cytosol, committing the cell to apoptosis by activating a cascade of cysteinyl-aspartate specific proteases (caspases). We are only beginning to understand how the choice between mitophagy and the activation of caspases is determined on the mitochondrial surface. Intriguingly in neurons, caspase activation also plays a non-apoptotic role in synaptic plasticity. Here we review the current knowledge on the interplay between mitophagy and caspase activation with a special focus on the central nervous system.

The outcome of proximal fifth metatarsal fractures: redefining treatment strategies.

BACKGROUND: To optimize the treatment strategy and reduce treatment costs of proximal fifth metatarsal fractures, clinical and patient-reported outcome, and its determinants were addressed. METHODS: A retrospective adult cohort study including 152 proximal fifth metatarsal fractures: 121 nonoperatively and 31 operatively treated. In the operative group, 21 were zone 1 and 10 zone 2 fractures. Median follow-up was 37.5 (IQR 20.8-52.3) months with a minimal follow-up of 6 months. Twenty-three demographic, fracture, and treatment characteristics were assessed as well as the healthcare costs. Outcome was assessed using the patient files, anterior-posterior and oblique X-rays, foot function index (FFI), visual analog score (VAS), and SF-36 questionnaires. RESULTS: The median FFI, physical SF-36, and VAS scores did not significantly differ between nonoperatively and operatively treated patients. The FFI and physical SF-36 were predominantly affected by a history of mobility impairment and pre-existent cardiovascular diseases, whereas mental SF-36 correlated significantly with higher ASA-score. Overall complication rate was 5.9% (4.1 vs. 12.9%; p = 0.065, nonoperative vs. operative, respectively). Nonunion was recorded in only one (nonoperatively) treated patient. The total healthcare costs for operative treatment were 4.2 times higher compared to nonoperative treatment (€1960 vs. €463 per patient, respectively). CONCLUSION: Overall, the clinical and patient-reported outcome was good. The foot function and quality of life were mainly affected by comorbidity, rather than fracture and treatment-related variables. Although nonoperatively treated patients indicated decreased mental quality of life, our study indicates that proximal fifth metatarsal fractures can safely be treated nonoperatively without the risk of nonunion, with fewer complications and lower healthcare costs. LEVEL OF EVIDENCE: 3.

Persistent STAT5 activation in myeloid neoplasms recruits p53 into gene regulation.

STAT (Signal Transducer and Activator of Transcription) transcription factors are constitutively activated in most hematopoietic cancers. We previously identified a target gene, LPP/miR-28 (LIM domain containing preferred translocation partner in lipoma), induced by constitutive activation of STAT5, but not by transient cytokine-activated STAT5. miR-28 exerts negative effects on thrombopoietin receptor signaling and platelet formation. Here, we demonstrate that, in transformed hematopoietic cells, STAT5 and p53 must be synergistically bound to chromatin for induction of LPP/miR-28 transcription. Genome-wide association studies show that both STAT5 and p53 are co-localized on the chromatin at 463 genomic positions in proximal promoters. Chromatin binding of p53 is dependent on persistent STAT5 activation at these proximal promoters. The transcriptional activity of selected promoters bound by STAT5 and p53 was significantly changed upon STAT5 or p53 inhibition. Abnormal expression of several STAT5-p53 target genes (LEP, ATP5J, GTF2A2, VEGFC, NPY1R and NPY5R) is frequently detected in platelets of myeloproliferative neoplasm (MPN) patients, but not in platelets from healthy controls. In conclusion, persistently active STAT5 can recruit normal p53, like in the case of MPN cells, but also p53 mutants, such as p53 M133K in human erythroleukemia cells, leading to pathologic gene expression that differs from canonical STAT5 or p53 transcriptional programs.

Thin conductive diamond films as beam intensity monitors for soft x-ray beamlines.

Quantitative analysis of X-ray absorption and dichroism data requires knowledge of the beamline photon flux during the measurements. We show that thin conductive (B-doped) diamond thin films can be an alternative to the widely used gold meshes for monitoring the beam intensity of soft X-ray beamlines in situ. Limited by the carbon extended x-ray absorption fine structure oscillations, the diamond films become applicable beginning from about 600 eV photon energy, where the important transition metal edges and the rare-earth edges are found. The 100 nm and 250 nm thick free-standing diamond films were grown and tested against standard gold meshes in real-life dichroism experiments performed at beamline ID08 of the European Synchrotron Radiation Facility, Grenoble, France. Quantitative agreement was found between the two experimental data sets. The films feature an extremely high transmission of about 90% and, at the same time, yield a sufficiently strong and clean reference signal. Furthermore, the thin films do not affect the shape of the transmitted beam. X-rays passing mesh-type monitors are subject to diffraction effects, which widen the beam and become particularly disturbing for small beamsizes in the micrometer range.

Long-term physical functioning and its association with somatic comorbidity and comorbid depression in patients with established rheumatoid arthritis: a longitudinal study.

OBJECTIVE: To describe long-term physical functioning and its association with somatic comorbidity and comorbid depression in patients with established rheumatoid arthritis (RA). METHODS: Longitudinal data over a period of 11 years were collected from 882 patients with RA at study inclusion. Patient-reported outcomes were collected in 1997, 1998, 1999, 2002, and 2008. Physical functioning was measured with the Health Assessment Questionnaire and the physical component summary score of the Short Form 36 health survey. Somatic comorbidity was measured by a questionnaire including 12 chronic diseases. Comorbid depression was measured with the Center for Epidemiologic Studies Depression Scale. We distinguished 4 groups of patients based on comorbidity at baseline. RESULTS: Seventy-two percent of the patients at baseline were women. The mean ± SD age was 59.3 ± 14.8 years and the median disease duration was 5.0 years (interquartile range 2.0-14.0 years). For the total group of patients with RA, physical functioning improved over time. Patients with somatic comorbidity, comorbid depression, or both demonstrated worse physical functioning than patients without comorbidity at all data collection points. Both groups with comorbid depression had the lowest scores. Only patients with both somatic comorbidity and comorbid depression showed significantly less improvement in physical functioning over time. CONCLUSION: Both somatic comorbidity and comorbid depression were negatively associated with physical functioning during an 11-year followup period. Furthermore, their combination seems to be especially detrimental to physical functioning over time. These results emphasize the need to take somatic comorbidity and comorbid depression into account in the screening and treatment of patients with RA.

Piezoelectric actuated micro-resonators based on the growth of diamond on aluminum nitride thin films.

Unimorph heterostructures based on piezoelectric aluminum nitride (AlN) and diamond thin films are highly desirable for applications in micro- and nanoelectromechanical systems. In this paper, we present a new approach to combine thin conductive boron-doped as well as insulating nanocrystalline diamond (NCD) with sputtered AlN films without the need for any buffer layers between AlN and NCD or polishing steps. The zeta potentials of differently treated nanodiamond (ND) particles in aqueous colloids are adjusted to the zeta potential of AlN in water. Thereby, the nucleation density for the initial growth of diamond on AlN can be varied from very low (10(8) cm(-2)), in the case of hydrogen-treated ND seeding particles, to very high values of 10(11) cm(-2) for oxidized ND particles. Our approach yielding high nucleation densities allows the growth of very thin NCD films on AlN with thicknesses as low as 40 nm for applications such as microelectromechanical beam resonators. Fabricated piezo-actuated micro-resonators exhibit enhanced mechanical properties due to the incorporation of boron-doped NCD films. Highly boron-doped NCD thin films which replace the metal top electrode offer Young's moduli of more than 1000 GPa.

Clinical evaluation of the Sorin Xtra(R) Autotransfusion System.

The performance of the Sorin Xtra® Autotransfusion System (ATS) was studied in 62 patients undergoing coronary artery bypass grafting. Blood was collected intraoperatively and washed using three different wash sets in 4 groups. Both collected and washed blood were analysed for hemoglobin levels and hematocrit, concentrations of proteins, albumin, heparin and plasma free hemoglobin (PFH) were determined, erythrocytes, platelets and leukocytes were counted. Hematocrit measurements of the Xtra® were compared with laboratory measurements to study the accuracy of the Xtra® hematocrit sensor. In addition, the red blood cell recovery rate and elimination rates were calculated to evaluate the clinical performance of the Xtra®. The Xtra® ATS produced a volume of concentrated red blood cells with an average hematocrit from 58% to 63%, depending on the size of the bowl and the chosen default program. In all bowl sizes and programs, the Xtra® Hct-out measurement underestimated the CELL-DYN measurement by approximately 15%. The calculated recovery rates for red blood cells (RBC) in the 4 groups ranged from 86.7% to 91.6%. Elimination rates were calculated in each group for proteins (96.8-99.2%), albumin (96.4-98.7%), plasma free hemoglobin (83.6-91.2%), heparin (98.8-99.9%), platelets (82.4-94.3%) and white blood cells (28.6-42.3%). The Xtra® ATS can be appealing for its performance by producing high hematocrit levels in the washed RBC volume, while keeping RBC recovery rate at the same high level (≈ 90%) as in its predecessor, the Electa® Autotransfusion System.

Sub-cycle switch-on of ultrastrong light-matter interaction.

Controlling the way light interacts with material excitations is at the heart of cavity quantum electrodynamics (QED). In the strong-coupling regime, quantum emitters in a microresonator absorb and spontaneously re-emit a photon many times before dissipation becomes effective, giving rise to mixed light-matter eigenmodes. Recent experiments in semiconductor microcavities reached a new limit of ultrastrong coupling, where photon exchange occurs on timescales comparable to the oscillation period of light. In this limit, ultrafast modulation of the coupling strength has been suggested to lead to unconventional QED phenomena. Although sophisticated light-matter coupling has been achieved in all three spatial dimensions, control in the fourth dimension, time, is little developed. Here we use a quantum-well waveguide structure to optically tune light-matter interaction from weak to ultrastrong and turn on maximum coupling within less than one cycle of light. In this regime, a class of extremely non-adiabatic phenomena becomes observable. In particular, we directly monitor how a coherent photon population converts to cavity polaritons during abrupt switching. This system forms a promising laboratory in which to study novel sub-cycle QED effects and represents an efficient room-temperature switching device operating at unprecedented speed.

Oral mucosal versus cutaneous sensory testing: a review of the literature.

The innervation of skin and oral mucosa plays a major physiological role in exteroception. It also has a clinical interest as illustrated by sensory changes after neurosurgical procedures. These sensory changes often rely only on the patients' subjective reports, although objective assessments are possible. This review compares the neurophysiological features of the trigeminal sensory pathways with those of cutaneous sensory innervation. In this review, three receptor groups will be discussed: mechanoreceptors, thermoreceptors and nociceptors. Differences between receptors in the glabrous skin, the hairy skin and the oral mucosa will be highlighted. Sensory testing devices have been developed to quantify psychophysiological parameters such as the threshold level for receptor activation upon mechanical stimulation, but such devices have been merely developed to determine the threshold of skin receptors (tactile, thermal). Later on, some have been adapted to suit the particularities of the oral environment. This review attempts to compare the available literature on test devices for oral versus cutaneous tactile function. It summarizes what is common or rather particular to the devices used to study either cutaneous or oral receptors.

Small intestinal morphology and disaccharidase activities in early-weaned piglets fed a diet containing spray-dried porcine plasma.

The purpose of this study was to test whether dietary spray-dried porcine plasma (SDPP) in early-weaned piglets prevents small intestinal villus atrophy by trophic or protective activity. Fifty-four weaned, 18-day-old piglets were used to determine the effect of dietary SDPP on small intestinal villus length, crypt depth, enterocyt mitotic activity and brush border enzyme activities during the first week after weaning. The piglets were offered a diet containing either 8% SDPP or 8% casein. At 2 and 7 days after weaning, piglets were anaesthetized to provide samples of the small intestinal wall and killed immediately afterwards. There were no differences in daily gain and daily feed intake between the two dietary treatments. At day 2 after weaning, all piglets showed a marked reduction in villus height when compared with baseline values. In all piglets, small intestinal enterocyte mitotic activity had decreased by day 2 and was increased again on day 7. There were no significant effects of dietary SDPP on small intestinal villus length, crypt depth and enterocyt mitotic activity. This indicates that SDPP has no trophic effect on the small intestinal mucosa and that it does not protect against the damaging effect on the small intestinal villi that is associated with the process of weaning. There was no effect of SDPP on lactase-, sucrase- or maltase-specific activities that are a measure of the digestive function of the small intestine. It can be concluded that SDPP versus casein has no effect on small intestinal morphology and disaccharidase activities in early weaned piglets kept under low infection pressure.

Trigeminal somatosensory evoked potentials in humans.

The recording of somatosensory evoked potentials (SEPs) is a non-invasive routine clinical testing procedure in neurology. For trigeminal nerve stimulation, however, SEPs have not received a widespread clinical attention. A variety of protocols and procedures have been used to record trigeminal SEPs (TSEPs). Differences encountered include the stimulation mode, site and frequency, the recording electrode position and data acquisition parameters. This has resulted in a diversity of recorded TSEP signals, making comparisons almost impossible. The general picture shows a number of short latency waves (within 3 ms) of peripheral origin, followed by at least two longer latency waves (12-15 ms and 19-22 ms). Furthermore, potential waves with a very long latency (> 100 ms) follow when the response is produced by painful stimulation. The origin of the long and very long latency waves is still a matter of debate. In order to allow reliable data interpretation and comparisons between the outcome of different studies, a standardized protocol should be applied for TSEP recordings. By providing an overview, this paper aims to mark a step forward in the harmonization of TSEP protocols with respect to the neural processes of interest. Further studies should also encounter the potential application of other neuroimaging techniques, such as functional magnetic resonance imaging or positron emission tomography, preferably in combination with TSEP recordings.

Sensations and trigeminal somatosensory-evoked potentials elicited by electrical stimulation of endosseous oral implants in humans.

The perception of bipolar electrical stimuli through implants was studied. The stimuli were delivered to permucosal oral endosseous implants in 15 individuals, who then reported tapping to beating sensations. In 10 out of the 15, these stimuli evoked clearly distinguishable potentials in the averaged electroencephalograms. The most prominent scalp potential was a positive wave with a latency between 18 and 25 ms, often preceded by a negative wave with a latency around 12-17 ms. In contrast, when a motor response was elicited by stimulation of the lip, a shorter latency wave around 8-11 ms was found additionally, indicating that the former-mentioned waves represent a true sensory response and not an artefact of myogenous origin. Furthermore, topical anaesthesia of the gingiva surrounding the implants in six individuals had little effect on the sensory responses. This evidence excluded peri-implant mucosal innervation as the origin of the perception and of the somatosensory-evoked waves elicited by the electrical stimulation of the oral implants. To the best of our knowledge, for the first time a sensation (osseoperception) has been elicited by electrical stimulation of endosseous oral implants and correlated with simultaneously recorded trigeminal somatosensory-evoked potentials (TSEPs).

Diagnosis of ulnar neuropathy: a new approach.

Conventional electrodiagnosis used to detect an ulnar neuropathy at the elbow depends on accurate determination of ulnar nerve length across this segment. We present a new approach, using the difference in latency of the compound nerve action potentials (CNAPs) of the ulnar and median nerves elicited by stimulation at the wrist and recorded 10 cm above the elbow. Sixty normal controls were examined in order to determine the normal upper limit (1.4 ms) of the difference in CNAP latency of the ulnar and the median nerves (Dlat index). Values obtained in 10 patients with ulnar nerve lesions are discussed. This test was shown to be both sensitive and specific, was independent of ulnar nerve length, and was easy to perform.

Value of somatosensory and motor evoked potentials in predicting arm recovery after a stroke.

OBJECTIVES: Prediction of motor recovery in the arm in patients with stroke is generally based on clinical examination. However, neurophysiological measures may also have a predictive value. The aims of this study were to assess the role of somatosensory (SSEPs) and motor (MEPs) evoked potentials in the prediction of arm motor recovery and to determine whether these measures added further predictive information to that gained from clinical examination. METHODS: Sixty four patients who had had a stroke and presented with obvious motor deficit of the arm were examined in terms of three clinical variables (motor performance, muscle tone, and overall disability) and for SSEPs and MEPs. Clinical and neurophysiological examinations were done at entry to the study (2 to 5 weeks poststroke), and at about 2 months after stroke. Further clinical follow up was conducted at 6 and 12 months after stroke. RESULTS: Neurophysiological measures made in the acute phase were of little use alone in predicting motor recovery of the arm at 2, 6, and 12 months after stroke. At 2 months, the absence of SSEPs and MEPs indicated a very poor outcome. Conversely, if the responses were preserved, a great variation in motor outcome was found. Multiple regression analysis showed that the addition of SSEPs and MEPs to the clinical examination increased the possibility of predicting arm recovery in the long term. In the acute phase, the combination of the motor score and SSEPs were best able to predict outcome. The long term outcome based on variables taken at 2 months, was best predicted through incorporating the three clinical measures and MEPs. CONCLUSIONS: Neurophysiological measures alone are of limited value in predicting long term outcome. However, predictive accuracy is substantially improved through the combined use of both of these measures and clinical variables.

[Vectorial competence of Glossina tachinoides Westwood and Glossina palpalis gambiensis Vanderplank infected by Trypanosoma brucei brucei EATRO 1125]. / Compétence vectorielle des Glossina tachinoides Westwood et Glossina palpalis gambiensis Vanderplank infectées simultanément par Trypanosoma brucei brucei EATRO 1125.

The vectorial competence (VC) of teneral (less than 32 h) Glossina tachinoides Westwood and G. palpalis gambiensis Vanderplank, fed simultaneously on a guinea-pig infected with Trypanosoma brucei brucei EATRO 1125, was assessed. Statistical analysis of the experimental results revealed that female G. tachinoides had a significantly higher midgut infection rate than males. Such a sex-related difference was not observed in G. p. gambiensis. Male G. p. gambiensis had higher midgut infection rates than male G. tachinoides. The metacyclic index did not differ between both subspecies, although G. p. gambiensis showed relatively more metacyclic infections than G. tachinoides. A global VC of 0.0242 and 0.0483 was found for G. tachinoides and G. p. gambiensis, respectively. VC did not differ significantly either between sexes or between the two species. However, G. tachinoides more rapidly infected the feeding host than G. p. gambiensis. In all infected flies, the procyclic index value was superior to the metacylic index value, suggesting that the infection is established by an ascending origin. Both large and slender parts of the salivary glands were constantly infected. Longitudinally dividing trypomastigotes of unequal length have been observed in the alimentary canal of the flies.