Super-Resolution Imaging of Subcortical White Matter using Stochastic Optical Reconstruction Microscopy (STORM) and Super-Resolution Optical Fluctuation Imaging (SOFI).

AIMS: The spatial resolution of light microscopy is limited by the wavelength of visible light (the "diffraction limit", approximately 250 nm). Resolution of sub-cellular structures, smaller than this limit, is possible with super resolution methods such as stochastic optical reconstruction microscopy (STORM) and super-resolution optical fluctuation imaging (SOFI). We aimed to resolve subcellular structures (axons, myelin sheaths and astrocytic processes) within intact white matter using STORM and SOFI. METHODS: Standard cryostat-cut sections of subcortical white matter from donated human brain tissue and from adult rat and mouse brain were labelled using standard immunohistochemical markers (neurofilament-H, myelin associated glycoprotein, GFAP). Image sequences were processed for STORM (effective pixel size 8-32 nm) and for SOFI (effective pixel size 80 nm). RESULTS: In human, rat and mouse subcortical white matter high quality images for axonal neurofilaments, myelin sheaths and filamentous astrocytic processes were obtained. In quantitative measurements, STORM consistently underestimated width of axons and astrocyte processes (compared with electron microscopy measurements). SOFI provided more accurate width measurements, though with somewhat lower spatial resolution than STORM. CONCLUSIONS: Super resolution imaging of intact cryo-cut human brain tissue is feasible. For quantitation, STORM can under-estimate diameters of thin fluorescent objects. SOFI is more robust. The greatest limitation for super-resolution imaging in brain sections is imposed by sample preparation. We anticipate that improved strategies to reduce autofluorescence and to enhance fluorophore performance will enable rapid expansion of this approach. [232 words] This article is protected by copyright. All rights reserved

Improved corrosion resistance of plasma carbon coated NiTi orthopedic materials

Nickel titanium (NiTi) alloys are useful in orthopedic applications because of their super-elastic properties and shape memory effects. However, when NiTi is used for a prolonged period of time, harmful Ni ions can leach out into the surrounding body fluid inside a human body, and so it is important to design a method to impede the out-diffusion of nickel from the materials into the biological medium. We aim at producing a barrier to mitigate the release of Ni ions during normal use. Carbon coatings have been shown to possess excellent bio-compatibility and good mechanical strength. In this work, amorphous hydrogenated DLC films with a graded C/NiTi interface were fabricated by plasma immersion ion implantation & deposition (PU & D) to provide such a barrier layer on NiTi. The elemental depth profiles and film thickness were determined by X-ray photoelectron spectroscopy (XPS) whereas the surface morphology was evaluated using atomic force microscopy (AFM). The film structure was studied by X-ray diffraction (XRD) and Raman spectroscopy. The corrosion resistance of the film was investigated using electrochemical tests based on ASTM G5-94. Compared to the control sample, the corrosion potential of the sample with the carbon coating changes from -250 to -50 mV and the film breakdown potential increases from 250 to 1200 mV. The corrosion current also diminishes from 10-6 to 10-7 A. The simulated body fluid (SBF) solutions after the electrochemical test were analyzed for Ni concentrations by inductively-coupled plasma mass spectrometry (ICPMS) and that data show that a much smaller amount of Ni has been released from the treated sample surface compared to the untreated control sample surface. Our results thus indicate that the deposited DLC film is effective in retarding the release of Ni ions from the bulk materials and more superior corrosion resistance is achieved based on our tests in a simulated fluid medium and at human body temperature.published_or_final_versio

Suppression of nickel out-diffusion from porous nickel-titanium shape memory alloy by plasma immersion ion implantation

Summary form only given. Porous nickel titanium is a promising material for medical application not only because of its super elasticity and shape memory effect but also the porous structure which may enhance bone growth due to the increased surface area. It is thus especially suitable for bone tissue in-growth and fixation of biomedical implants. However, like its dense counterpart, Ni leaching from the materials causes health concern. Thus, in order to suppress Ni diffusion from the materials to body fluids and tissues in humans, a diffusion barrier or similar structure must be introduced. In this work, we produced this diffusion barrier layer by oxygen or nitrogen plasma immersion ion implantation (PIII). In vitro tests were conducted by immersing the plasma-treated NiTi into simulated body fluid (SBF) at 37plusmn0.5degC for 5 weeks and the resulting SBF was analyzed for Ni and Ti using inductively-coupled plasma mass spectrometry (ICMPS). Our results show that Ni leaching is significantly mitigated by both nitrogen and oxygen PIII.published_or_final_versio

Nickel suppression in Ni-Ti alloys by plasma immersion ion implantation surface treatment: New materials for orthopaedic implantation

Conference Theme: Spinal Motion Segment: From Basic Science to Clinical Applicationpublished_or_final_versio

The Population Impact of a Large School-Based Influenza Vaccination Campaign

The optimal vaccination strategy to mitigate the impact of influenza epidemics is unclear. In 2005, a countywide school-based influenza vaccination campaign was launched in Knox County, Tennessee (population 385,899). Approximately 41% and 48% of eligible county children aged 5-17 years were immunized with live attenuated influenza vaccine before the 2005-2006 and 2006-2007 influenza seasons, respectively. We sought to determine the population impact of this campaign.Laboratory-confirmed influenza data defined influenza seasons. We calculated the incidence of medically attended acute respiratory illness attributable to influenza in Knox and Knox-surrounding counties (concurrent controls) during consecutive seasons (5 precampaign and 2 campaign seasons) using negative binomial regression and rate difference methods. Age-stratified analyses compared the incidence of emergency department (ED) visits and hospitalizations attributable to influenza.During precampaign seasons, estimated ED visit rates attributable to influenza were 12.39 (95% CI: 10.34-14.44) per 1000 Knox children aged 5-17 years and similar in Knox-surrounding counties. During the campaign seasons, annual Knox influenza-associated ED visit rates declined relative to rates in Knox-surrounding counties: rate ratios 0.55 (95% CI: 0.27-0.83) and 0.70 (95% CI: 0.56-0.84) for the first and second campaign seasons, respectively. Overall, there were about 35% or 4.86 per 1000 fewer influenza-associated ED visits among Knox County children aged 5-17 years attributable to the campaign. No significant declines in Knox compared to surrounding counties were detected for influenza associated ED visits in children aged <5 years, all adults combined or selected adult age subgroups, although power for these analyses was limited. Alternate rate-difference analyses yielded consistent results.Vaccination of approximately 45% of Knox school-aged children with influenza vaccine was associated with a 35% annual reduction (4.86 per 1000) in ED visit rates attributable to influenza. Higher vaccination coverage and/or larger studies would be needed to determine whether similar interventions have indirect benefits in other age groups

Depression literacy among Australians of Chinese-speaking background in Melbourne, Australia

<p>Abstract</p> <p>Background</p> <p>This study investigated the knowledge of depression and preference for professional help, medications and treatment methods among Australians of Chinese-speaking background, and the perceptions of this population of the causes of mental illness.</p> <p>Methods</p> <p>Adopting a cluster convenience sampling method, the study recruited 200 Chinese-speaking subjects from four major areas in metropolitan Melbourne where many Chinese live. The respondents were presented with a vignette describing an individual with depression and then asked questions to assess their understanding of depression and preference for professional help, medications and treatment methods. A comparative approach was used to compare the findings with those of a previous study of the mental health literacy of Australian and Japanese adults.</p> <p>Results</p> <p>Compared to the Australian and Japanese samples, a much lower percentage of Chinese-speaking Australians (14%) could correctly identify major depression described in the vignette, and a higher percentage believed that counseling professionals could be helpful. Higher percentages of those who believed that close family members could be helpful were found in the Chinese-speaking Australian and Japanese samples, and these two groups also expressed more uncertainty about the usefulness or harmfulness of certain medications compared to the Australian sample. Higher percentages of respondents in both the Chinese-speaking Australian and the Australian sample considered "lifestyle changes" to be helpful compared to the Japanese sample. In the Chinese-speaking sample, 30%, 17.4%, 33% and 27% of the respondents rated "traditional Chinese medicine doctors," "Chinese herbal medications," "taking Chinese nutritional foods/supplements" and "<it>qiqong</it>" as helpful. Many perceived "changing <it>fungshui</it>" and "traditional Chinese worship" to be harmful. Regarding the perception of causes of mental illness, items related to psychosocial perspectives including "life stress" and "interpersonal conflict" were rated highly by the respondents, whereas traditional beliefs including "punishment for misdeeds conducted by ancestors" and "demon possession" had the lowest ratings.</p> <p>Conclusions</p> <p>Campaigns to increase the mental health literacy of Chinese-speaking Australians are needed. The abovementioned socially and culturally driven beliefs need to be taken into consideration in the development of culturally relevant education programs.</p

Essential versus accessory aspects of cell death: recommendations of the NCCD 2015

Cells exposed to extreme physicochemical or mechanical stimuli die in an uncontrollable manner, as a result of their immediate structural breakdown. Such an unavoidable variant of cellular demise is generally referred to as ‘accidental cell death’ (ACD). In most settings, however, cell death is initiated by a genetically encoded apparatus, correlating with the fact that its course can be altered by pharmacologic or genetic interventions. ‘Regulated cell death’ (RCD) can occur as part of physiologic programs or can be activated once adaptive responses to perturbations of the extracellular or intracellular microenvironment fail. The biochemical phenomena that accompany RCD may be harnessed to classify it into a few subtypes, which often (but not always) exhibit stereotyped morphologic features. Nonetheless, efficiently inhibiting the processes that are commonly thought to cause RCD, such as the activation of executioner caspases in the course of apoptosis, does not exert true cytoprotective effects in the mammalian system, but simply alters the kinetics of cellular demise as it shifts its morphologic and biochemical correlates. Conversely, bona fide cytoprotection can be achieved by inhibiting the transduction of lethal signals in the early phases of the process, when adaptive responses are still operational. Thus, the mechanisms that truly execute RCD may be less understood, less inhibitable and perhaps more homogeneous than previously thought. Here, the Nomenclature Committee on Cell Death formulates a set of recommendations to help scientists and researchers to discriminate between essential and accessory aspects of cell death

Modulators of Cytoskeletal Reorganization in CA1 Hippocampal Neurons Show Increased Expression in Patients at Mid-Stage Alzheimer's Disease

During the progression of Alzheimer's disease (AD), hippocampal neurons undergo cytoskeletal reorganization, resulting in degenerative as well as regenerative changes. As neurofibrillary tangles form and dystrophic neurites appear, sprouting neuronal processes with growth cones emerge. Actin and tubulin are indispensable for normal neurite development and regenerative responses to injury and neurodegenerative stimuli. We have previously shown that actin capping protein beta2 subunit, Capzb2, binds tubulin and, in the presence of tau, affects microtubule polymerization necessary for neurite outgrowth and normal growth cone morphology. Accordingly, Capzb2 silencing in hippocampal neurons resulted in short, dystrophic neurites, seen in neurodegenerative diseases including AD. Here we demonstrate the statistically significant increase in the Capzb2 expression in the postmortem hippocampi in persons at mid-stage, Braak and Braak stage (BB) III-IV, non-familial AD in comparison to controls. The dynamics of Capzb2 expression in progressive AD stages cannot be attributed to reactive astrocytosis. Moreover, the increased expression of Capzb2 mRNA in CA1 pyramidal neurons in AD BB III-IV is accompanied by an increased mRNA expression of brain derived neurotrophic factor (BDNF) receptor tyrosine kinase B (TrkB), mediator of synaptic plasticity in hippocampal neurons. Thus, the up-regulation of Capzb2 and TrkB may reflect cytoskeletal reorganization and/or regenerative response occurring in hippocampal CA1 neurons at a specific stage of AD progression

Cell Free Expression of hif1α and p21 in Maternal Peripheral Blood as a Marker for Preeclampsia and Fetal Growth Restriction

Preeclampsia, a severe unpredictable complication of pregnancy, occurs in 6% of pregnancies, usually in the second or third trimester. The specific etiology of preeclampsia remains unclear, although the pathophysiological hallmark of this condition appears to be an inadequate blood supply to the placenta. As a result of the impaired placental blood flow, intrauterine growth restriction (IUGR) and consequential fetal oxidative stress may occur. Consistent with this view, pregnancies complicated by preeclampsia and IUGR are characterized by up-regulation of key transcriptional regulators of the hypoxic response including, hif1α and as well as p53 and its target genes. Recently, the presence of circulating cell-free fetal RNA has been documented in maternal plasma. We speculated that pregnancies complicated by preeclampsia and IUGR, will be associated with an abnormal expression of p53 and/or hif1α related genes in the maternal plasma. Maternal plasma from 113 singleton pregnancies (72 normal and 41 complicated pregnancies) and 19 twins (9 normal and 10 complicated pregnancies) were collected and cell free RNA was extracted. The expression of 18 genes was measured by one step real-time RT-PCR and was analyzed for prevalence of positive/negative expression levels. Results indicate that, among the genes examined, cell free plasma expressions of p21 and hif1α were more prevalent in pregnancies complicated by hypoxia and/or IUGR (p<0.001). To conclude, we present in this manuscript data to support the association between two possible surrogate markers of hypoxia and common complications of pregnancy. More work is needed in order to implement these findings in clinical practice