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Tlio following flight-director displays were compared in tho laboratory on a simple aircraft simulator under controlled conditions of noiso and ambiorit illumination: (a) Streaming Lights which presented errors in the flight of the ‘ aircraft ’ by tho direction and rate of apparent movement of streaming lights; (b) ’ Barber's Poles ’ which presented errors by the direction and rate of translational movement of a white helical strip along a black, rotating polo; (c) Flashing Lights attached to the simulator, or to a helmet worn by tho subject, which presented errors in each dimension by the position and rate of flash of a single flashing light; (d) l.L.S. Meter which was intended to represent, a Zero-Reader display and presented errors by the position of two pointers mounted at right angles to each other. All these displays were designed to present navigational information to a pilot while he was scanning the outside world, and all except the last presented tho information in peripheral vision. In continuous tracking, the time off target with Flashing Lights or tho I.L.S. Meter was about a quarter of the time off target with either tho Streaming Lights or tho Barber's Poles. In correcting sudden errors, Flashing Lights on tho Helmet gave quicker responses than any other display which was investigated. This was presumed to bo the result of the high attention-getting value and the immediate directional indication of the signals. The weakness of Flashing Lights on tho Helmet, which also applied to the Barber's Poles and Streaming Lights, was in presenting information on the size of errors. The I.L.S. Meter was the best display in this respect, although it did not always attract the man's attention as soon as it indicated an error. The combination of Flashing Lights on tho Helmet and tho I.L.S. Meter produced tho quickest corrections recorded during the experiments. Reaction time to signals presented on a central display increased about 40 per cent when attention had to be paid to any of the flight-director displays. Tho size of tho increase was about the same whether simulated control of tho aircraft was carried out or not while performing the central task. This suggests that it was tho need to attend to the additional channel of information, rather than simultaneous demands for action, which interfered with the central task. Performance with Flashing Lights on the Helmet and Streaming Lights showed only a small and not statistically significant adverse effect from occasional rotation of the head and eyes of 70°. Sideways movements of tho head altered the angle subtended at the subject's eye by the Barber's Poles mounted horizontally fore and aft to display information on altitude. This changed the apparent rate of movement of the display and the apparent display-to-cantrol ratio, and thus caused the subject to miss small errors occasionally, or make control movements of the wrong size. In addition, with tho Barber's Poles the display-control directional relationships changed as attention was directed from one end of the azimuth display to the other. This could occur in on aircraft when the pilot rotated his head and eyes, and might be dangerous. [ABSTRACT FROM PUBLISHER]

Oligodendrocytes (OLs) of the central nervous system require iron for proteolipid biosynthesis during the myelination process. Although most heme is found complexed to hemoglobin in red blood cells, surprisingly, we found that Slc48a1, encoding the heme transporter Hrg1, is expressed at higher levels in OLs than any other cell type in rodent and humans. We confirmed in situ that Hrg1 is expressed in OLs but not their precursors (OPCs) and found that Hrg1 proteins in CNS white matter co-localized within myelin sheaths. In older Hrg1 null mutant mice we observed reduced expression of myelin associated glycoprotein (Mag) and ultrastructural myelin defects reminiscent of Mag-null animals, suggesting myelin adhesion deficiency. Further, we confirmed reduced myelin iron levels in Hrg1 null animals in vivo, and show that OLs in vitro can directly import both the fluorescent heme analogue ZnMP and heme itself, which rescued iron deficiency induced inhibition of OL differentiation in a heme-oxidase-dependent manner. Together these findings indicate OL Hrg1 encodes a functional heme transporter required for myelin integrity., (© 2024 The Author(s). GLIA published by Wiley Periodicals LLC.)

Alterations to cerebral white matter tracts have been associated with cognitive decline in aging and Alzheimer's disease (AD). In particular, the fornix has been implicated as especially vulnerable given that it represents the primary outflow tract of the hippocampus. Despite this, little work has focused on the fornix using a potential early marker of white matter degeneration-myelin water fraction (MWF; an in vivo marker of myelin content). Therefore, we sought to (1) clarify associations between MWF in the fornix and memory functioning, and (2) examine whether fornix MWF relates to memory performance above and beyond hippocampal volume and conventional imaging measures of white matter that may not be as specific to alterations in myelin content. Forty nondemented older adults (mean age = 72.9 years) underwent an MRI exam and neuropsychological assessment. Multicomponent driven equilibrium single pulse observation of T1 and T2 (mcDESPOT) was used to quantify fornix MWF and diffusion tensor imaging (DTI) was used to measure fornix fractional anisotropy (FA). Adjusting for age, sex, education, and vascular risk factors, linear regression models revealed that, lower fornix MWF was significantly associated with poorer memory functioning (β = 0.405, p = .007) across our sample of older adults. Notably, fornix MWF remained a significant predictor of memory functioning (β = 0.380, p = .015) even after adjusting for fornix DTI FA and hippocampal volume (in addition to the above covariates). Given the observed associations between myelin and memory in older adults without dementia, MWF may be a useful early marker of dementia risk., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

The mediobasal hypothalamus (MBH; arcuate nucleus of the hypothalamus [ARH] and median eminence [ME]) is a key nutrient sensing site for the production of the complex homeostatic feedback responses required for the maintenance of energy balance. Here, we show that refeeding after an overnight fast rapidly triggers proliferation and differentiation of oligodendrocyte progenitors, leading to the production of new oligodendrocytes in the ME specifically. During this nutritional paradigm, ME perineuronal nets (PNNs), emerging regulators of ARH metabolic functions, are rapidly remodeled, and this process requires myelin regulatory factor (Myrf) in oligodendrocyte progenitors. In genetically obese ob/ob mice, nutritional regulations of ME oligodendrocyte differentiation and PNN remodeling are blunted, and enzymatic digestion of local PNN increases food intake and weight gain. We conclude that MBH PNNs are required for the maintenance of energy balance in lean mice and are remodeled in the adult ME by the nutritional control of oligodendrocyte differentiation., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Although cerebral blood flow (CBF) alterations are associated with Alzheimer's disease (AD), CBF patterns across prodromal stages of AD remain unclear. Therefore, we investigated patterns of regional CBF in 162 Alzheimer's Disease Neuroimaging Initiative participants characterized as cognitively unimpaired (CU; n  = 80), objectively-defined subtle cognitive decline (Obj-SCD; n  = 31), or mild cognitive impairment (MCI; n  = 51). Arterial spin labeling MRI quantified regional CBF in a priori regions of interest: hippocampus, inferior temporal gyrus, inferior parietal lobe, medial orbitofrontal cortex, and rostral middle frontal gyrus. Obj-SCD participants had increased hippocampal and inferior parietal CBF relative to CU and MCI participants and increased inferior temporal CBF relative to MCI participants. CU and MCI groups did not differ in hippocampal or inferior parietal CBF, but CU participants had increased inferior temporal CBF relative to MCI participants. There were no CBF group differences in the two frontal regions. Thus, we found an inverted-U pattern of CBF signal across prodromal AD stages in regions susceptible to early AD pathology. Hippocampal and inferior parietal hyperperfusion in Obj-SCD may reflect early neurovascular dysregulation, whereby higher CBF is needed to maintain cognitive functioning relative to MCI participants, yet is also reflective of early cognitive inefficiencies that distinguish Obj-SCD from CU participants.

Although the cerebral cortex is organized into six excitatory neuronal layers, it is unclear whether glial cells show distinct layering. In the present study, we developed a high-content pipeline, the large-area spatial transcriptomic (LaST) map, which can quantify single-cell gene expression in situ. Screening 46 candidate genes for astrocyte diversity across the mouse cortex, we identified superficial, mid and deep astrocyte identities in gradient layer patterns that were distinct from those of neurons. Astrocyte layer features, established in the early postnatal cortex, mostly persisted in adult mouse and human cortex. Single-cell RNA sequencing and spatial reconstruction analysis further confirmed the presence of astrocyte layers in the adult cortex. Satb2 and Reeler mutations that shifted neuronal post-mitotic development were sufficient to alter glial layering, indicating an instructive role for neuronal cues. Finally, astrocyte layer patterns diverged between mouse cortical regions. These findings indicate that excitatory neurons and astrocytes are organized into distinct lineage-associated laminae.

Recent interest in astrocyte activation states has raised the fundamental question of how these cells, normally essential for synapse and neuronal maintenance, become pathogenic. Here, we show that activation of the unfolded protein response (UPR), specifically phosphorylated protein kinase R-like endoplasmic reticulum (ER) kinase (PERK-P) signaling-a pathway that is widely dysregulated in neurodegenerative diseases-generates a distinct reactivity state in astrocytes that alters the astrocytic secretome, leading to loss of synaptogenic function in vitro. Further, we establish that the same PERK-P-dependent astrocyte reactivity state is harmful to neurons in vivo in mice with prion neurodegeneration. Critically, targeting this signaling exclusively in astrocytes during prion disease is alone sufficient to prevent neuronal loss and significantly prolongs survival. Thus, the astrocyte reactivity state resulting from UPR over-activation is a distinct pathogenic mechanism that can by itself be effectively targeted for neuroprotection., Competing Interests: Declaration of Interests The authors declare no competing interests. O.J.F. is now an employee of AstraZeneca., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Multiple sclerosis (MS) is a neuroinflammatory disease with a relapsing-remitting disease course at early stages, distinct lesion characteristics in cortical grey versus subcortical white matter and neurodegeneration at chronic stages. Here we used single-nucleus RNA sequencing to assess changes in expression in multiple cell lineages in MS lesions and validated the results using multiplex in situ hybridization. We found selective vulnerability and loss of excitatory CUX2-expressing projection neurons in upper-cortical layers underlying meningeal inflammation; such MS neuron populations exhibited upregulation of stress pathway genes and long non-coding RNAs. Signatures of stressed oligodendrocytes, reactive astrocytes and activated microglia mapped most strongly to the rim of MS plaques. Notably, single-nucleus RNA sequencing identified phagocytosing microglia and/or macrophages by their ingestion and perinuclear import of myelin transcripts, confirmed by functional mouse and human culture assays. Our findings indicate lineage- and region-specific transcriptomic changes associated with selective cortical neuron damage and glial activation contributing to progression of MS lesions.

Ageing causes a decline in tissue regeneration owing to a loss of function of adult stem cell and progenitor cell populations 1 . One example is the deterioration of the regenerative capacity of the widespread and abundant population of central nervous system (CNS) multipotent stem cells known as oligodendrocyte progenitor cells (OPCs) 2 . A relatively overlooked potential source of this loss of function is the stem cell 'niche'-a set of cell-extrinsic cues that include chemical and mechanical signals 3,4 . Here we show that the OPC microenvironment stiffens with age, and that this mechanical change is sufficient to cause age-related loss of function of OPCs. Using biological and synthetic scaffolds to mimic the stiffness of young brains, we find that isolated aged OPCs cultured on these scaffolds are molecularly and functionally rejuvenated. When we disrupt mechanical signalling, the proliferation and differentiation rates of OPCs are increased. We identify the mechanoresponsive ion channel PIEZO1 as a key mediator of OPC mechanical signalling. Inhibiting PIEZO1 overrides mechanical signals in vivo and allows OPCs to maintain activity in the ageing CNS. We also show that PIEZO1 is important in regulating cell number during CNS development. Thus we show that tissue stiffness is a crucial regulator of ageing in OPCs, and provide insights into how the function of adult stem and progenitor cells changes with age. Our findings could be important not only for the development of regenerative therapies, but also for understanding the ageing process itself.

During early human pregnancy the uterine mucosa transforms into the decidua, into which the fetal placenta implants and where placental trophoblast cells intermingle and communicate with maternal cells. Trophoblast-decidual interactions underlie common diseases of pregnancy, including pre-eclampsia and stillbirth. Here we profile the transcriptomes of about 70,000 single cells from first-trimester placentas with matched maternal blood and decidual cells. The cellular composition of human decidua reveals subsets of perivascular and stromal cells that are located in distinct decidual layers. There are three major subsets of decidual natural killer cells that have distinctive immunomodulatory and chemokine profiles. We develop a repository of ligand-receptor complexes and a statistical tool to predict the cell-type specificity of cell-cell communication via these molecular interactions. Our data identify many regulatory interactions that prevent harmful innate or adaptive immune responses in this environment. Our single-cell atlas of the maternal-fetal interface reveals the cellular organization of the decidua and placenta, and the interactions that are critical for placentation and reproductive success.

Many lines of evidence suggest that the Parkinson's disease (PD)-related protein α-synuclein (α-SYN) can propagate from cell to cell in a prion-like manner. However, the cellular mechanisms behind the spreading remain elusive. Here, we show that human astrocytes derived from embryonic stem cells actively transfer aggregated α-SYN to nearby astrocytes via direct contact and tunneling nanotubes (TNTs). Failure in the astrocytes' lysosomal digestion of excess α-SYN oligomers results in α-SYN deposits in the trans-Golgi network followed by endoplasmic reticulum swelling and mitochondrial disturbances. The stressed astrocytes respond by conspicuously sending out TNTs, enabling intercellular transfer of α-SYN to healthy astrocytes, which in return deliver mitochondria, indicating a TNT-mediated rescue mechanism. Using a pharmacological approach to inhibit TNT formation, we abolished the transfer of both α-SYN and mitochondria. Together, our results highlight the role of astrocytes in α-SYN cell-to-cell transfer, identifying possible pathophysiological events in the PD brain that could be of therapeutic relevance. SIGNIFICANCE STATEMENT Astrocytes are the major cell type in the brain, yet their role in Parkinson's disease progression remains elusive. Here, we show that human astrocytes actively transfer aggregated α-synuclein (α-SYN) to healthy astrocytes via direct contact and tunneling nanotubes (TNTs), rather than degrade it. The astrocytes engulf large amounts of oligomeric α-SYN that are subsequently stored in the trans-Golgi network region. The accumulation of α-SYN in the astrocytes affects their lysosomal machinery and induces mitochondrial damage. The stressed astrocytes respond by sending out TNTs, enabling intercellular transfer of α-SYN to healthy astrocytes. Our findings highlight an unexpected role of astrocytes in the propagation of α-SYN pathology via TNTs, revealing astrocytes as a potential target for therapeutic intervention., (Copyright © 2017 Rostami et al.)

α-Synuclein is a presynaptic protein associated to Parkinson's disease, which is unstructured when free in the cytoplasm and adopts α helical conformation when bound to vesicles. After decades of intense studies, α-Synuclein physiology is still difficult to clear up due to its interaction with multiple partners and its involvement in a pletora of neuronal functions. Here, we looked at the remarkably neglected interplay between α-Synuclein and microtubules, which potentially impacts on synaptic functionality. In order to identify the mechanisms underlying these actions, we investigated the interaction between purified α-Synuclein and tubulin. We demonstrated that α-Synuclein binds to microtubules and tubulin α2β2 tetramer; the latter interaction inducing the formation of helical segment(s) in the α-Synuclein polypeptide. This structural change seems to enable α-Synuclein to promote microtubule nucleation and to enhance microtubule growth rate and catastrophe frequency, both in vitro and in cell. We also showed that Parkinson's disease-linked α-Synuclein variants do not undergo tubulin-induced folding and cause tubulin aggregation rather than polymerization. Our data enable us to propose α-Synuclein as a novel, foldable, microtubule-dynamase, which influences microtubule organisation through its binding to tubulin and its regulating effects on microtubule nucleation and dynamics.

Induced pluripotent stem cells (iPSCs) are becoming an important source of pre-clinical models for research focusing on neurodegeneration. They offer the possibility for better understanding of common and divergent pathogenic mechanisms of brain diseases. Moreover, iPSCs provide a unique opportunity to develop personalized therapeutic strategies, as well as explore early pathogenic mechanisms, since they rely on the use of patients' own cells that are otherwise accessible only post-mortem, when neuronal death-related cellular pathways and processes are advanced and adaptive. Neurodegenerative diseases are in majority of unknown cause, but mutations in specific genes can lead to familial forms of these diseases. For example, mutations in the superoxide dismutase 1 gene lead to the motor neuron disease amyotrophic lateral sclerosis (ALS), while mutations in the SNCA gene encoding for alpha-synuclein protein lead to familial Parkinson's disease (PD). The generations of libraries of familial human ALS iPSC lines have been described, and the iPSCs rapidly became useful models for studying cell autonomous and non-cell autonomous mechanisms of the disease. Here we report the generation of a comprehensive library of iPSC lines of familial PD and an associated synucleinopathy, multiple system atrophy (MSA). In addition, we provide examples of relevant neural cell types these iPSC can be differentiated into, and which could be used to further explore early disease mechanisms. These human cellular models will be a valuable resource for identifying common and divergent mechanisms leading to neurodegeneration in PD and MSA., Competing Interests: The authors declare no conflict of interest.

Concentration of viable cell populations in suspension is of interest for several clinical and pre-clinical applications. Here, we report that microfluidic acoustophoresis is an effective method to efficiently concentrate live and viable cells with high target purity without any need for protein fluorescent labeling using antibodies or over-expression. We explored the effect of the acoustic field acoustic energy density and systematically used different protocols to induce apoptosis or cell death and then determined the efficiency of live and dead cell separation. We used the breast cancer cell line MCF-7, the mouse neuroblastoma N2a as well as human embryonic stem cells (hESCs) to demonstrate that this method is gentle and can be applied to different cell populations. First, we induced cell death by means of high osmotic shock using a high concentration of PBS (10×), the protein kinase inhibitor staurosporine, high concentrations of dimethyl sulfoxide (DMSO, 10%), and finally, cell starvation. In all the methods employed, we successfully induced cell death and were able to purify and concentrate the remaining live cells using acoustophoresis. Importantly, the concentration of viable cells was not dependent on a specific cell type. Further, we demonstrate that different death inducing stimuli have different effects on the intrinsic cell properties and therefore affect the efficiency of the acoustophoretic separation.

Purpose: The aim of this study was to research the associations between noise (ambient and activity noise) and objective metrics of teachers' voices in real working environments (i.e., classrooms)., Method: Thirty-two female and 8 male teachers from 14 elementary schools were randomly selected for the study. Ambient noise was measured during breaks in unoccupied classrooms and, likewise, the noise caused by pupils' activity during lessons. Voice samples were recorded before and after a working day. Voice variables measured were sound pressure level (voice SPL), fundamental frequency, jitter, shimmer, and the tilt of the sound spectrum slope (alpha ratio)., Results: The ambient noise correlated most often with the fundamental frequency of men and voice SPL, whereas activity noise correlated with the alpha ratio and perturbation values. Teachers working in louder ambient noise spoke more loudly before work than those working in lower noise levels. Voice variables generally changed less during work among teachers working in loud activity noise than among those working in lower noise levels., Conclusions: Ambient and activity noises affect teachers' voice use. Under loud ambient noise teachers seem to speak habitually loudly, and under loud activity noise teachers' ability to react to loading deteriorates.

Phonation into glass tubes ('resonance tubes'), keeping the free end of the tube in water, has been a frequently used voice therapy method in Finland and more recently also in other countries. The purpose of this exploratory study was to investigate what effects tube phonation with and without water has on the larynx. Two participants were included in the study. The methods used were high-speed imaging, electroglottographic observations of vocal fold vibrations, and measurements of oral pressure during tube phonation. Results showed that the fluctuation in the back pressure during tube phonation in water altered the vocal fold vibrations. In the high-speed imaging, effects were found in the open quotient and amplitude variation of the glottal opening. The open quotient increased with increasing water depth (from 2 cm to 6 cm). A modulation effect by the water bubbles on the vocal fold vibrations was seen both in the high-speed glottal area tracings and in the electroglottography signal. A second experiment revealed that the increased average oral pressure was largely determined by the water depth. The increased open quotient can possibly be explained by an increased abduction of the vocal folds and/or a reduced transglottal pressure. The back pressure of the bubbles also modulates glottal vibrations with a possible 'massage' effect on the vocal folds. This effect and the well-defined average pressure increase due to the known water depth are different from those of other methods using a semi-occluded vocal tract.

The associations between voice ergonomic risk factors in 40 classrooms and the acoustic parameters of 40 schoolteachers' voices were investigated. The risk factors assessed were connected to participants' working practices, working postures, and the indoor air quality in their workplaces. The teachers recorded spontaneous speech and sustained /a/ before and after a working day. Fundamental frequency, sound pressure level, the slope of the spectrum, perturbation, and harmonic-to-noise ratio were analysed. The results showed that the more the voice ergonomic risk factors were involved, the louder the teachers' voices became. Working practices correlated most often with the acoustic parameters; associations were found especially before a working day. The results suggest that a risky voice ergonomic environment affects voice production.

In this study, we sought evidence for alpha-synuclein (ASYN) expression in oligodendrocytes, as a possible endogenous source of ASYN to explain its presence in glial inclusions found in multiple system atrophy (MSA) and Parkinson's disease (PD). We identified ASYN in oligodendrocyte lineage progenitors isolated from the rodent brain, in oligodendrocytes generated from embryonic stem cells, and in induced pluripotent stem cells produced from fibroblasts of a healthy individual and patients diagnosed with MSA or PD, in cultures in vitro. Notably, we observed a significant decrease in ΑSYN during oligodendrocyte maturation. Additionally, we show the presence of transcripts in PDGFRΑ/CD140a(+) cells and SOX10(+) oligodendrocyte lineage nuclei isolated by FACS from rodent and human healthy and diseased brains, respectively. Our work identifies ASYN in oligodendrocyte lineage cells, and it offers additional in vitro cellular models that should provide significant insights of the functional implication of ASYN during oligodendrocyte development and disease., (Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.)

Astrocytes play a critical role during the development and the maintenance of the CNS in health and disease. Yet, their lack of accessibility from fetuses and from the brain of diseased patients has hindered our understanding of their full implication in developmental and pathogenic processes. Human pluripotent stem cells (PSCs) are an alternative source to obtain large quantities of astrocytes in vitro, for mechanistic studies of development and disease. However, these studies often require highly pure populations of astrocytes, which are not always achieved, depending on the PSC lines and protocols used. Here, we describe the generation and characterization of human PSC reporter lines expressing TagRFP driven by the ABC1D region of the human GFAP promoter, as new cellular model for generating homogenous population of astrocytes generated from CNS regionally defined PSC-derived neural progenitors. GFA(ABC1D)::TagRFP-expressing astrocytes can be purified by fluorescent-activated cell sorting and maintain a bright expression for several additional weeks. These express canonical astrocyte markers NF1A, S100β, CX43, GLAST, GS and CD44. These new cellular models, from which highly pure populations of fluorescence-expressing astrocytes can be obtained, provide a new platform for studies where pure or fluorescently labeled astrocyte populations are necessary, for example to assess pro-inflammatory cytokine and chemokine release in response to specific treatment, and uptake and degradation of fluorescently labeled pathogenic proteins, as reported in this study., (Copyright © 2015. Published by Elsevier B.V.)

Objectives: The purpose of this cross-sectional study was to determine the prevalence of hoarseness in children attending the first or second grade of primary school and to explore possible background factors for hoarseness in children., Methods: The participants were 217 children, aged 6-10 years, from 10 different schools. Questionnaires were filled in by the parents and the teachers of the children and voice samples were recorded. The voice samples from the children were perceptually evaluated by eight trained listeners and intra- and inter-rater reliability was calculated. Additionally, the parents and teachers were in the questionnaires asked to rate the children's voices. Connections between background factors and voice quality were explored., Results: Both the intra- and inter-rater reliability for the trained listeners were relatively high and significant. The prevalence of hoarseness for the whole group was 12.0% as judged by the trained listeners. For girls, the prevalence of hoarseness was 7.8% and for boys 15.8%. A lower teacher rating of degree of maturity correlated significantly with the voice quality. Additionally, there was a significant negative correlation between the amount of talking at home and voice quality. For girls, heavy voice use as an infant correlated significantly with voice quality. For boys, being the youngest sibling correlated significantly with voice quality., Conclusions: The results from the present study indicate that more attention should be paid to hoarseness in children and that background factors should be further explored., (Copyright © 2015 The Voice Foundation. Published by Elsevier Inc. All rights reserved.)

The cellular hallmarks of Parkinson's disease (PD) are the loss of nigral dopaminergic neurons and the formation of α-synuclein-enriched Lewy bodies and Lewy neurites in the remaining neurons. Based on the topographic distribution of Lewy bodies established after autopsy of brains from PD patients, Braak and coworkers hypothesized that Lewy pathology primes in the enteric nervous system and spreads to the brain, suggesting an active retrograde transport of α-synuclein (the key protein component in Lewy bodies), via the vagal nerve. This hypothesis, however, has not been tested experimentally thus far. Here, we use a human PD brain lysate containing different forms of α-synuclein (monomeric, oligomeric and fibrillar), and recombinant α-synuclein in an in vivo animal model to test this hypothesis. We demonstrate that α-synuclein present in the human PD brain lysate and distinct recombinant α-synuclein forms are transported via the vagal nerve and reach the dorsal motor nucleus of the vagus in the brainstem in a time-dependent manner after injection into the intestinal wall. Using live cell imaging in a differentiated neuroblastoma cell line, we determine that both slow and fast components of axonal transport are involved in the transport of aggregated α-synuclein. In conclusion, we here provide the first experimental evidence that different α-synuclein forms can propagate from the gut to the brain, and that microtubule-associated transport is involved in the translocation of aggregated α-synuclein in neurons.

Synucleinopathies, characterized by intracellular aggregation of α-synuclein protein, share a number of features in pathology and disease progression. However, the vulnerable cell population differs significantly between the disorders, despite being caused by the same protein. While the vulnerability of dopamine cells in the substantia nigra to α-synuclein over-expression, and its link to Parkinson's disease, is well studied, animal models recapitulating the cortical degeneration in dementia with Lewy-bodies (DLB) are much less mature. The aim of this study was to develop a first rat model of widespread progressive synucleinopathy throughout the forebrain using adeno-associated viral (AAV) vector mediated gene delivery. Through bilateral injection of an AAV6 vector expressing human wild-type α-synuclein into the forebrain of neonatal rats, we were able to achieve widespread, robust α-synuclein expression with preferential expression in the frontal cortex. These animals displayed a progressive emergence of hyper-locomotion and dysregulated response to the dopaminergic agonist apomorphine. The animals receiving the α-synuclein vector displayed significant α-synuclein pathology including intra-cellular inclusion bodies, axonal pathology and elevated levels of phosphorylated α-synuclein, accompanied by significant loss of cortical neurons and a progressive reduction in both cortical and striatal ChAT positive interneurons. Furthermore, we found evidence of α-synuclein sequestered by IBA-1 positive microglia, which was coupled with a distinct change in morphology. In areas of most prominent pathology, the total α-synuclein levels were increased to, on average, two-fold, which is similar to the levels observed in patients with SNCA gene triplication, associated with cortical Lewy body pathology. This study provides a novel rat model of progressive cortical synucleinopathy, showing for the first time that cholinergic interneurons are vulnerable to α-synuclein over-expression. This animal model provides a powerful new tool for studies of neuronal degeneration in conditions of widespread cortical α-synuclein pathology, such as DLB, as well an attractive model for the exploration of novel biomarkers.

Objectives/hypothesis: Stress reaction provokes changes in the body involving cardiovascular alterations, autonomic reactions, neuroendocrine and immunologic as well as psychoneuroimmunologic changes. Both the primary and secondary effect of stress reaction may be of consequence for vocal function. The purpose of this questionnaire study was to determine the effect of stress symptoms on the occurrence of vocal symptoms. The study also aimed at investigating whether a possible effect was different for men and women., Methods: A total number of 1728 participants completed a questionnaire concerning speech, language, and voice. Six vocal symptoms and four possible stress symptoms were included in the questionnaire., Results: There was a significant association between stress symptoms and the occurrence of vocal symptoms. The occurrence of muscle tension or a lump in the throat was significant regarding all the four possible stress symptoms. There were also significant results concerning gender difference. All vocal symptoms and two of four stress symptoms were more common among women., Conclusions: Physical changes caused by the stress reaction may result in vocal symptoms as those presented in the study. Both vocal symptoms and stress symptoms were more common among women. Stress should preferably be acknowledged as a risk factor containing and possibly entailing a number of physiological, psychological, and behavioral symptoms affecting the voice negatively., (Copyright © 2013 The Voice Foundation. Published by Mosby, Inc. All rights reserved.)

Objectives: The aim of the study was to investigate the connections between voice ergonomic risk factors found in classrooms and voice-related problems in teachers., Methods: Voice ergonomic assessment was performed in 39 classrooms in 14 elementary schools by means of a Voice Ergonomic Assessment in Work Environment--Handbook and Checklist. The voice ergonomic risk factors assessed included working culture, noise, indoor air quality, working posture, stress, and access to a sound amplifier. Teachers from the above-mentioned classrooms reported their voice symptoms, respiratory tract diseases, and completed a Voice Handicap Index (VHI)., Results: The more voice ergonomic risk factors found in the classroom the higher were the teachers' total scores on voice symptoms and VHI. Stress was the factor that correlated most strongly with voice symptoms. Poor indoor air quality increased the occurrence of laryngitis., Conclusions: Voice ergonomics were poor in the classrooms studied and voice ergonomic risk factors affected the voice. It is important to convey information on voice ergonomics to education administrators and those responsible for school planning and taking care of school buildings., (Copyright © 2012 The Voice Foundation. Published by Mosby, Inc. All rights reserved.)

Objective: The aim of the study was to investigate if voice ergonomic risk factors in classrooms correlated with acoustic parameters of teachers' voice production., Methods: The voice ergonomic risk factors in the fields of working culture, working postures and indoor air quality were assessed in 40 classrooms using the Voice Ergonomic Assessment in Work Environment - Handbook and Checklist. Teachers (32 females, 8 males) from the above-mentioned classrooms recorded text readings before and after a working day. Fundamental frequency, sound pressure level (SPL) and the slope of the spectrum (alpha ratio) were analyzed., Results: The higher the number of the risk factors in the classrooms, the higher SPL the teachers used and the more strained the males' voices (increased alpha ratio) were. The SPL was already higher before the working day in the teachers with higher risk than in those with lower risk., Conclusion: In the working environment with many voice ergonomic risk factors, speakers increase voice loudness and use more strained voice quality (males). A practical implication of the results is that voice ergonomic assessments are needed in schools., (Copyright © 2013 S. Karger AG, Basel.)

The pharmaceutical industry, particularly the small molecule domain, faces unprecedented challenges of escalating costs, high attrition as well as increasing competitive pressure from other companies and from new treatment modes such as biological products. In other industries, process improvement approaches, such as Lean Sigma, have delivered benefits in speed, quality and cost of delivery. Examining the medicinal chemistry contributions to the iterative improvement process of design-make-test-analyse from a Lean Sigma perspective revealed that major improvements could be made. Thus, the cycle times of synthesis, as well as compound analysis and purification, were reduced dramatically. Improvements focused on team, rather than individual, performance. These new ways of working have consequences for staff engagement, goals, rewards and motivation, which are also discussed.

Comparatively little is known about directed motility of environmental bacteria to common aromatic pollutants. Here, by expressing different parts of a (methyl)phenol-degradative pathway and the use of specific mutants, we show that taxis of Pseudomonas putida towards (methyl)phenols is dictated by its ability to catabolize the aromatic compound. Thus, in contrast to previously described chemoreceptor-mediated chemotaxis mechanisms towards benzoate, naphthalene and toluene, taxis in response to (methyl)phenols is mediated by metabolism-dependent behaviour. Here we show that P. putida differentially expresses three Aer-like receptors that are all polar-localized through interactions with CheA, and that inactivation of the most abundant Aer2 protein significantly decreases taxis towards phenolics. In addition, the participation of a sensory signal transduction protein composed of a PAS, a GGDEF and an EAL domain in motility towards these compounds is demonstrated. The results are discussed in the context of the versatility of metabolism-dependent coupling and the necessity for P. putida to integrate diverse metabolic signals from its native heterogeneous soil and water environments.