Complete Loss Of Signal Research Articles

In Vivo Recording of Neural and Behavioral Correlates of Anesthesia Induction, Reversal, and Euthanasia in Cephalopod Molluscs.

Cephalopod molluscs are among the most behaviorally and neurologically complex invertebrates. As they are now included in research animal welfare regulations in many countries, humane and effective anesthesia is required during invasive procedures. However, currently there is no evidence that agents believed to act as anesthetics produce effects beyond immobility. In this study we demonstrate, for the first time, that two of the most commonly used agents in cephalopod general anesthesia, magnesium chloride and ethanol, are capable of producing strong and reversible blockade of afferent and efferent neural signal; thus they are genuine anesthetics, rather than simply sedating agents that render animals immobile but not insensible. Additionally, we demonstrate that injected magnesium chloride and lidocaine are effective local anesthetic agents. This represents a considerable advance for cephalopod welfare. Using a reversible, minimally invasive recording procedure, we measured activity in the pallial nerve of cuttlefish (Sepia bandensis) and octopus (Abdopus aculeatus, Octopus bocki), during induction and reversal for five putative general anesthetic and two local anesthetic agents. We describe the temporal relationship between loss of behavioral responses (immobility), loss of efferent neural signal (loss of “consciousness”) and loss of afferent neural signal (anesthesia) for general anesthesia, and loss of afferent signal for local anesthesia. Both ethanol and magnesium chloride were effective as bath-applied general anesthetics, causing immobility, complete loss of behavioral responsiveness and complete loss of afferent and efferent neural signal. Cold seawater, diethyl ether, and MS-222 (tricaine) were ineffective. Subcutaneous injection of either lidocaine or magnesium chloride blocked behavioral and neural responses to pinch in the injected area, and we conclude that both are effective local anesthetic agents for cephalopods. Lastly, we demonstrate that a standard euthanasia protocol—immersion in isotonic magnesium chloride followed by surgical decerebration—produced no behavioral response and no neural activity during surgical euthanasia. Based on these data, we conclude that both magnesium chloride and ethanol can function as general anesthetic agents, and lidocaine and magnesium chloride can function as local anesthetic agents for cephalopod molluscs.

A Structural and Functional Role for Disulfide Bonds in a Class II Hydrophobin.

Hydrophobins are multifunctional, highly surface active proteins produced in filamentous fungi and can be identified by eight conserved cysteine residues, which form four disulfide bridges. These proteins can be subdivided into two classes based on their hydropathy profiles, solubility, and structures formed upon interfacial assembly. Here, we probe the structural and functional roles of disulfide bonds for a class II hydrophobin in different interfacial contexts by reducing its disulfides with 1,4-dithiothreitol and blocking the free thiols with iodoacetamide and then examining the protein secondary structure, emulsification capability, hydrophobic surface wetting, and solution self-assembly. Changes in circular dichroism spectra upon reduction and blocking of disulfides are consistent with an increase in the level of random coil secondary structure. Emulsification of octane in water using reduced and unreduced forms of class II hydrophobin showed a substantial loss of emulsification ability without disulfides and stable emulsion formation for hydrophobin with disulfides. Additionally, water contact angle measurements performed on polytetrafluoroethylene treated with solutions of reduced and unreduced hydrophobin showed efficient wetting of the hydrophobic surface for unreduced samples only. Lastly, Förster resonance energy transfer (FRET) was used to assess the role of disulfides in self-assembly in solution, and near complete loss of the FRET signal is consistent with a model in which solution self-assembly does not occur after reduction and blocking of the disulfides. From this, we conclude that, in contrast to class I hydrophobins, the disulfides of this class II hydrophobin are required for protein structural stability, surface activity at both liquid-liquid and solid-liquid interfaces, and solution self-assembly.

Abstract CT076: Multicenter phase II study of nivolumab in previously treated patients with recurrent and metastatic non-keratinizing nasopharyngeal carcinoma - Mayo clinic Phase 2 Consortium P2C-MN026, NCI9742, NCT02339558

Abstract Background: Endemic nasopharyngeal carcinoma (NPC) refers to non-keratinizing (NK) subtypes harboring Epstein Barr virus (EBV) that frequently express programmed death-ligand 1 (PD-L1). We recently reported ≥30% of NPC harbor MHC class 1 gene alterations in a whole genome study. Method: We investigated the activity of nivolumab (NIVO) & relationship with PD-L1 & HLA expression (exp) in tumor cells (TC) & tumor infiltrating cells (TIL). Eligible patient (pt)s had to fail ≥ 1 prior line of platinum-based chemo. NIVO (3mg/kg IV, q2 weeks) was given until disease progression (PD). Archived tumors were analyzed for PD-L1 (DAKO) & HLA mutations (indicated by loss of HLA-A &/or B exp). Result: A total of 45 pts were enrolled across 11 sites from Oct 2015 to June 2016. Overall, 43 and 44 pts were evaluable respectively for response rate (RR, RECIST) & adverse events (AE). The median age was 57 years, 77% pts were males & 84% were Asians. The median number of prior chemo was 3 (range 1-9 lines). Of the 43 evaluable pts, there were 8 confirmed (19%, 95% CI: 8-33%) & 1 unconfirmed partial response (PR), 17 PD (40%), 14 stable disease (SD, 33%) & 3 pts were not assessed. One pt (MD015-027) had an unconfirmed clinical CR in an occipital metastasis (met). Pts received a median of 3 cycles of NIVO (range 1 - 14). Median overall survival has not been reached yet. Of the 44 pts evaluable for AE, 8 (18.2%) had grade 3 or higher treatment-related AEs & 1 pt died of sepsis. The most common grade 3 or higher AEs were increase in alanine/aspartate aminotransferase (n=3), & 1 pt respectively for hyponatremia, neutropenia, fatigue & diarrhea. Of the 21 tumor samples analyzed, 11/21 (52%) had >1% PD-L1 expressed mainly in TC. Table 1 outlines relationship between RR & PD-L1 exp. All pts with PR had >5% PD-L1 exp & one pt (34003-022) with CR (nasopharynx) & PR (lung mets) had the highest exp (80%). Majority of pts with PD had <1% PD-L1 exp tumors. One primary progressor (34003-032) had a PD-L1 +ve tumor harboring a complete loss of HLA-B signal (TC), but intact signal in TILs & stromal cells. Conclusion: NIVO is active in this heavily pre-treated group of pts with NK-NPC and longer follow-up is needed for survival endpoints. Preliminary result suggests PD-L1 exp may predict benefit from NIVO. The analytical result of all 45 tumors will be presented. Table 1.PD-L1 exp and RECIST response in 21 ptsPD-L1%espRR%PRSDPD0-11<1%-265%24-10%--120%-1-30%11-80%1--Total498 Citation Format: Brigette B. Ma, Boon Cher Goh, Wan T. Lim, Kwok W. Lo, Edwin P. Hui, Jonathan W. Riess, Mark Agulnik, Alex Y. Chang, Julie A. Kish, Dean W. Lim, Douglas R. Adkins, Kevin J. Cullen, Barbara J. Gitlitz, Nathan R. Foster, Adam M. Pettinger, Sanna Mckinzie, Ka F. To, Brian Costello, Howard Streicher, Anthony T. Chan. Multicenter phase II study of nivolumab in previously treated patients with recurrent and metastatic non-keratinizing nasopharyngeal carcinoma - Mayo clinic Phase 2 Consortium P2C-MN026, NCI9742, NCT02339558 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr CT076. doi:10.1158/1538-7445.AM2017-CT076

Retroviral replicating vector-mediated gene therapy achieves long-term control of tumor recurrence and leads to durable anticancer immunity.

Background.Prodrug-activator gene therapy with Toca 511, a tumor-selective retroviral replicating vector (RRV) encoding yeast cytosine deaminase, is being evaluated in recurrent high-grade glioma patients. Nonlytic retroviral infection leads to permanent integration of RRV into the cancer cell genome, converting infected cancer cell and progeny into stable vector producer cells, enabling ongoing transduction and viral persistence within tumors. Cytosine deaminase in infected tumor cells converts the antifungal prodrug 5-fluorocytosine into the anticancer drug 5-fluorouracil, mediating local tumor destruction without significant systemic adverse effects.Methods.Here we investigated mechanisms underlying the therapeutic efficacy of this approach in orthotopic brain tumor models, employing both human glioma xenografts in immunodeficient hosts and syngeneic murine gliomas in immunocompetent hosts.Results.In both models, a single injection of replicating vector followed by prodrug administration achieved long-term survival benefit. In the immunodeficient model, tumors recurred repeatedly, but bioluminescence imaging of tumors enabled tailored scheduling of multicycle prodrug administration, continued control of disease burden, and long-term survival. In the immunocompetent model, complete loss of tumor signal was observed after only 1–2 cycles of prodrug, followed by long-term survival without recurrence for >300 days despite discontinuation of prodrug. Long-term survivors rejected challenge with uninfected glioma cells, indicating immunological responses against native tumor antigens, and immune cell depletion showed a critical role for CD4+ T cells.Conclusion.These results support dual mechanisms of action contributing to the efficacy of RRV-mediated prodrug-activator gene therapy: long-term tumor control by prodrug conversion-mediated cytoreduction, and induction of antitumor immunity.

UAV Navigation using Signals of Opportunity in Urban Environments: A Review

Abstract Novel Communication, Navigation and Surveillance (CNS) systems are currently developed targeting the Required Total System Performance (RTSP) levels. Within RTSP, it is essential to meet the Required Navigation Performance (RNP) in all flight phases, especially considering the Unmanned Aerial System (UAS) evolutions in the UAS Traffic Management (UTM) context. However, in dense urban environments characterized by tall buildings and complex structural variations, Global Navigation Satellite System (GNSS) is prone to data degradations or complete loss of signal due to multipath effects, interference or antenna obscuration. Furthermore, there is always a risk of jamming and spoofing of GNSS signals, with low cost civilian GNSS receivers being more vulnerable to a spoof attack. Therefore, a number of Signals of Opportunity (SoOP) techniques are explored to improve the RNP when Unmanned Aerial Vehicles (UAV) are employed in urban canyons. Electromagnetic signals found in urban environment including analogue/ digital radio, analogue/digital television, Wi-Fi, Global System for Mobile Communications (GSM) and Code Division Multiple Access (CDMA) based signals are considered to model the system performance parameters. Implementation methods for using Signals of Opportunity such as Angle of Arrival (AOA), Time of Arrival (TOA), Received Signal Strength (RSS) and Time Difference of Arrival (TDOA) are modeled and compared. Integration of SoOP techniques in novel low-cost Navigation and Guidance Systems (NGS) is also investigated. As SoOP were not initially designed for navigation purposes, no single source of SoOP for navigation can work in all environments and hence a SoOP source has to be selected based on specific requirements in the considered urban environment. Constraints of power and weight on the UAV besides hardware and software costs are also factors that are considered when selecting appropriate SoOP signal sources. Hence, there is a clear opportunity to provide considerable savings in both infrastructure as well as energy costs and thus providing a low-cost and low-volume integrated NGS for trusted aerial autonomous operations.

Retrospective Waveform Analysis of Transcranial Motor Evoked Potentials (MEP) to Identify Early Predictors of Impending Motor Deficits in Spinal Surgeries

ABSTRACTPurpose: Although there are guidelines analyzing transcranial motor evoked potentials (MEP) waveform criteria, they vary widely and are not applied universally during intraoperative neurophysiologic monitoring (IONM). The objective is to generate hypotheses to identify early and reliable MEP waveform characteristics prior to complete loss of MEP to predict impending motor spinal cord injuries during spinal surgeries. The ultimate goal is to enhance real-time feedback to prevent injury or detect reversible spinal cord damage.Methods: Fifteen true positive cases of persistent intraoperative MEP loss and new postoperative motor deficits were retrospectively identified from 2011 to 2013. Waveform characteristics of latency, amplitude, duration, phases, and area-under-the-curve (AUC) were measured, and an intraoperative spinal cord index (ISCI) was calculated for 5 traces prior to complete MEP loss. ISCI = [amplitude x duration x (phases+1) x AUC]/latency.Results: Out of 22 muscles in 15 cases, latency increased in 2, duration decreased in 12, amplitude decreased in 13, AUC decreased in 13, and ISCI decreased in 14. In 11 out of 15 cases (73%), ISCI dropped > 40% in at least one muscle before MEP were completely lost. Thirteen cases had concurrent somatosensory evoked potentials (SSEP) changes, 9 out of 13 had > 50% decrease in SSEP: 2 out of 9 changed before MEP, 5 out of 9 simultaneously, and 2 out of 9 after.Conclusions: In these cases of motor injury, smaller and simpler MEP waveforms preceded complete loss of signal. An ISCI 40% drop could be tested as a warning threshold for impending motor compromise in future prospective studies and lead to eventual standardization to predict irreversible postoperative deficits.

Serial in vivo imaging of transplanted allogeneic neural stem cell survival in a mouse model of amyotrophic lateral sclerosis

Neural stem cells (NSCs) are being investigated as a possible treatment for amyotrophic lateral sclerosis (ALS) through intraspinal transplantation, but no longitudinal imaging studies exist that describe the survival of engrafted cells over time. Allogeneic firefly luciferase-expressing murine NSCs (Luc+-NSCs) were transplanted bilaterally (100,000 cells/2μl) into the cervical spinal cord (C5) parenchyma of pre-symptomatic (63day-old) SOD1G93A ALS mice (n=14) and wild-type age-matched littermates (n=14). Six control SOD1G93A ALS mice were injected with saline. Mice were immunosuppressed using a combination of tacrolimus+sirolimus (1mg/kg each, i.p.) daily. Compared to saline-injected SOD1G93A ALS control mice, a transient improvement (p<0.05) in motor performance (rotarod test) was observed after NSC transplantation only at the early disease stage (weeks 2 and 3 post-transplantation). Compared to day one post-transplantation, there was a significant decline in bioluminescent imaging (BLI) signal in SOD1G93A ALS mice at the time of disease onset (71.7±17.9% at 4weeks post-transplantation, p<0.05), with a complete loss of BLI signal at endpoint (120day-old mice). In contrast, BLI signal intensity was observed in wild-type littermates throughout the entire study period, with only a 41.4±8.7% decline at the endpoint. In SOD1G93A ALS mice, poor cell survival was accompanied by accumulation of mature macrophages and the presence of astrogliosis and microgliosis. We conclude that the disease progression adversely affects the survival of engrafted murine Luc+-NSCs in SOD1G93A ALS mice as a result of the hostile ALS spinal cord microenvironment, further emphasizing the challenges that face successful cell therapy of ALS.

Viability-PCR Shows That NAAT Detects a High Proportion of DNA from Non-Viable Chlamydia trachomatis.

ObjectivesAccording to the current guidelines for laboratory diagnosis of sexually transmitted infections (STIs), nucleic acid amplification tests (NAATs) are the preferred diagnostic method for Chlamydia trachomatis (CT) infections. However, NAATs amplify the available target DNA without discriminating between DNA originating from viable or non-viable CT. Assessing CT viability will provide more insights in the clinical and public health relevance of a CT positive test result. The aim of this study was to technically validate and implement viability-PCR (V-PCR) to asses CT viability.MethodsTechnical validation of V-PCR was performed by the assessment of predefined viability ratios of CT. Samples were subjected to V-PCR which consisted of propidium monoazide (PMA) treatment prior to DNA extraction followed by quantitative PCR (qPCR) targeting the ompA gene for the detection of CT DNA. Finally, V-PCR was applied to vaginal swabs of 50 CT positive patients, as indicated by routine NAAT, collected at our outpatient STD clinics before antimicrobial treatment.ResultsTechnical validation of V-PCR showed that PMA treatment of heat-inactivated CT culture resulted in an almost complete loss of qPCR signal. PMA treated samples of the fresh viable CT culture showed no marked reduction of PCR signal, indicating that all DNA from viable CT could be detected. Applying V-PCR to clinical samples showed that in 36% of samples (18/50) less than 1% of CT DNA originated from viable bacteria.ConclusionsV-PCR showed to be a fast and easy method to assess CT viability in clinical samples, without the need of traditional challenging cell culture methods. Furthermore, V-PCR results of clinical samples have indicated that a substantial amount of the amplified CT DNA originated from non-viable cells. Although results might be influenced by cell death during transport, this study suggests that there is a potential overestimation of quantitative CT positivity by currently used NAATs.

Is Transthyretin a Regulator of Ubc9 SUMOylation?

Ageing and mutations of transthyretin (TTR), the thyroid hormones and retinol transporting protein lead to amyloidosis by destabilizing the structure of TTR. Because protein structure is regulated through posttranslational modifications, we investigated the Small Ubiquitin-like Modifier (SUMO)ylation of TTR. We chose the widely used Ubc9 fusion-directed SUMOylation system, which is based on a fusion of the SUMOylation substrate of interest with Ubc9, a sole SUMO conjugating enzyme. Surprisingly, despite our presumptions, we found that Ubc9 fused to TTR was SUMOylated at a unique set of lysine residues. Three unknown SUMOylation sites of Ubc9—K154, K18 and K65—were revealed by mass spectrometry (MS). The previously reported SUMOylation at K49 of Ubc9 was also observed. SUMOylation of the lysine residues of TTR fused to Ubc9 was hardly detectable. However, non-fused TTR was SUMOylated via trans-SUMOylation by Ubc9 fused to TTR. Interestingly, mutating the catalytic residue of Ubc9 fused to TTR did not result in complete loss of the SUMOylation signal, suggesting that Ubc9 linked to TTR is directly cross-SUMOylated by the SUMO-activating enzyme E1. Ubc9, TTR or fusion proteins composed of TTR and Ubc9 specifically affected the global SUMOylation of cellular proteins. TTR or Ubc9 alone increased global SUMOylation, whereas concomitant presence of TTR and Ubc9 did not further increase the amount of high-molecular weight (HMW) SUMO conjugates. Our data suggest that TTR may influence the SUMOylation of Ubc9, thereby altering signalling pathways in the cell.

Investigation of the Impact of Desorption Electrospray Ionization Sprayer Geometry on Its Performance in Imaging of Biological Tissue.

In this study, the impact of sprayer design and geometry on performance in desorption electrospray ionization mass spectrometry (DESI-MS) is assessed, as the sprayer is thought to be a major source of variability. Absolute intensity repeatability, spectral composition, and classification accuracy for biological tissues are considered. Marked differences in tissue analysis performance are seen between the commercially available and a lab-built sprayer. These are thought to be associated with the geometry of the solvent capillary and the resulting shape of the primary electrospray. Experiments with a sprayer with a fixed solvent capillary position show that capillary orientation has a crucial impact on tissue complex lipid signal and can lead to an almost complete loss of signal. Absolute intensity repeatability is compared for five lab-built sprayers using pork liver sections. Repeatability ranges from 1 to 224% for individual sprayers and peaks of different spectral abundance. Between sprayers, repeatability is 16%, 9%, 23%, and 34% for high, medium, low, and very low abundance peaks, respectively. To assess the impact of sprayer variability on tissue classification using multivariate statistical tools, nine human colorectal adenocarcinoma sections are analyzed with three lab-built sprayers, and classification accuracy for adenocarcinoma versus the surrounding stroma is assessed. It ranges from 80.7 to 94.5% between the three sprayers and is 86.5% overall. The presented results confirm that the sprayer setup needs to be closely controlled to obtain reliable data, and a new sprayer setup with a fixed solvent capillary geometry should be developed.

Recurrent laryngeal nerve injury with incomplete loss of electromyography signal during monitored thyroidectomy-evaluation and outcome.

During monitored thyroidectomy, a partially or completely disrupted point of nerve conduction on the exposed recurrent laryngeal nerve (RLN) indicates true electrophysiologic nerve injury. Complete loss of signal (LOS; absolute threshold value <100μV) at the end of operation often indicates a postoperative vocal cord (VC) palsy. However, the evaluation for the injured RLN with incomplete LOS and its functional outcome has not been well described. Three hundred twenty-three patients with 522 RLNs at risk who underwent standardized monitored thyroidectomy were enrolled. The RLN was routinely stimulated at the most proximal (R2p signal) and distal (R2d signal) ends of exposure after thyroid resection to determine if there was an injured point on the RLN. Pre- and postoperative VC function was routinely examined. Twenty-nine RLNs (5.6%) were detected with an injury point. Five nerves had complete LOS and other 24 nerves had incomplete LOS where the R2p/R2d reduction (% of amplitude reduction compared with proximal to distal RLN stimulation) ranged from 22 to 79%. Postoperative temporary VC palsy was noted in those five RLNs with complete LOS (final vagal signal, V2 < 100μV) and four RLNs with incomplete LOS (R2p/R2d reduction 62-79%; V2 181-490μV). In the remaining 20 nerves with R2p/R2d reduction ≤53% (V2 373-1623μV), all showed normal VC mobility. Overall, false negative results were found in two RLNs (0.4%) featuring unchanged V2 and R2p/R2d but developed VC palsy. Testing and comparing the R2p/R2d signal is a simple and useful procedure to evaluate RLN injury after its dissection and predict functional outcome. When the relative threshold value R2p/R2d reduction reaches over 60%, surgeon should consider the possibility of postoperative VC palsy.

Dynamics of loss and recovery of the nerve monitoring signal during thyroidectomy predict early postoperative vocal fold function.

The characteristics of segmental type 1 and global type 2 injuries to the recurrent laryngeal nerve (RLN) and the extent and dynamics of nerve recovery are poorly understood. This investigation of 785 patients who underwent thyroidectomy under continuous intraoperative nerve monitoring aimed at exploring the dynamics of loss and recovery of the nerve monitoring signal and its relationship to early postoperative vocal fold palsy. Persistent complete loss of signal and signal recovery <50% identified all (based on 12 and 4 patients with type 1 injuries) or most (based on 9 of 12 and 4 of 6 patients with global type 2 injuries) early unilateral vocal fold palsies. Signal recovery ≥50% (based on 7 patients) always signified normal vocal fold function. These data, including the observation that global type 2 injuries may entail less severe nerve injury, require validation in independent series before being adopted more widely. © 2015 Wiley Periodicals, Inc. Head Neck 38: E1144-E1151, 2016.

Transport Stream Analysis of a ISDB-T Signal Using an Embedded and Reconfigurable System

Quality management is an important criterion to evaluate broadcaster's transmission system integrity, which can be affected by radiofrequency signal impairments and by coding, multiplexing, and/or modulation errors. The international system integrated services digital broadcasting - terrestrial (ISDB-T) was designed to provide a reliable reception, even with interferences inherent to radio wave propagation. However, channel noise, as well as problems in the transmission system, may corrupt the transport stream (TS), causing image failures and, depending on the error level, even a complete loss of signal. This paper presents a supervisory system consisting of an ISDB-T front-end with spatial diversity of four branches, associated with a system-on-chip (SoC) board, in order to analyze the TS of any broadcaster, identifying transmission errors in real-time. The system continuously searches received data for impairments such as cyclic redundancy check - 32 and continuity counter errors, table/section intervals, and program clock reference jitter. The SoC comprises a field programmable gate array that includes a hard-wired processor, used to compute and show the analysis results.

Does intraoperative nerve monitoring reliably aid in staging of total thyroidectomies?

Demonstrate whether intraoperative nerve monitoring is an effective tool in staging bilateral thyroid and neck surgeries in cases of intraoperative injury to the recurrent laryngeal nerve on the side of initial dissection. We hypothesized that IONM provides reliable and appropriate feedback on the functional status of the RLN on side of initial dissection during total thyroidectomy and central neck surgery. Case series with planned data collection. All patients receiving total thyroidectomies or central neck surgeries were reviewed. The outcomes of patients treated whose procedures were staged based on intraoperative nerve monitoring are described. Ten (4.9%) of 206 procedures were staged based on unfavorable signal change including six patients with thyroid cancers and four with compressive substernal goiters. The overall signal drop in the 10 patients with unfavorable signal change was 63%. Three patients had complete loss of signal. In the other seven patients, the signal dropped by a mean of 48%. In the eight patients with laryngoscopy-proven vocal fold paresis, the signal dropped by a mean of 68%. Intraoperative nerve monitoring results accurately indicated postoperative ipsilateral vocal cord dysfunction with high reliability. Intraoperative nerve monitoring is a reliable tool in staging thyroid surgery. 4.

Delay to formalin fixation effect on HER2 test in breast cancer by dual-color silver-enhanced in situ hybridization (Dual-ISH).

HER2 status is an integral part of breast cancer management. One of the variables that negatively affects HER2 test is delay to formalin fixation (DFF). The purpose of this study is to determine the effect of progressive DFF on HER2-dual color in situ hybridization (Dual-ISH). Ten palpable invasive breast cancers were included in the study. For each case, the procured tumor was divided into 8 parts and consecutively fixed after 0, 10, 30 minutes, 1, 2, 4, and 8 hours; 1 section was kept in saline and stored overnight (ON) at 4°C. Two tissue microarray blocks were constructed. Then, HER2 was tested with Dual-ISH. HER2 and CEP17 signal and HER2/CEP17 ratio was calculated in 10 cells for each core. The percentage of cells that had complete loss of signal was calculated. The percentage of cells that had nuclear bubbling and/or background staining was also calculated. DFF had no statistically significant effect on HER2/CEP17 ratio, absolute number of HER2, or CEP17 signals. However, 1 case became falsely amplified in ON sample. The trend of the percentage of cells that lost signal with time of DFF was statistically significant (P<0.001), more notably after 1 hour of DFF. Nuclear bubbling and/or haze or dust was seen in 4 cases (P=0.009). These changes started to appear at 30 minutes to 1 hour DFF (P=0.025 and 0.003, respectively). DFF has no statistically significant effect on the HER2/CEP17 ratio tested by Dual-ISH. However, significant artifacts could occur in samples fixed after 1 hour of DFF time.

Effect of Oxidation on the Structure of Human Low- and High-Density Lipoproteins

This work presents a controlled study of low-density lipoprotein (LDL) and high-density lipoprotein (HDL) structural changes due to in vitro oxidation with copper ions. The changes were studied by small-angle x-ray scattering (SAXS) and dynamic light scattering (DLS) techniques in the case of LDL and by SAXS, DLS, and Z-scan (ZS) techniques in the case of HDL. SAXS data were analyzed with a to our knowledge new deconvolution method. This method provides the electron density profile of the samples directly from the intensity scattering of the monomers. Results show that LDL particles oxidized for 18 h show significant structural changes when compared to nonoxidized particles. Changes were observed in the electrical density profile, in size polydispersity, and in the degree of flexibility of the APO-B protein on the particle. HDL optical results obtained with the ZS technique showed a decrease of the amplitude of the nonlinear optical signal as a function of oxidation time. In contrast to LDL results reported in the literature, the HDL ZS signal does not lead to a complete loss of nonlinear optical signal after 18 h of copper oxidation. Also, the SAXS results did not indicate significant structural changes due to oxidation of HDL particles, and DLS results showed that a small number of oligomers formed in the sample oxidized for 18 h. All experimental results for the HDL samples indicate that this lipoprotein is more resistant to the oxidation process than are LDL particles.

Immobilisation of cell‐binding peptides on poly‐ε‐caprolactone (PCL) films: A comparative XPS study of two chemical surface functionalisation methods

Successful interaction between cells and biomaterial surfaces is crucial for biomedical devices, and incorporation of peptides such as RGD (Arg‐Gly‐Asp) at the polymer interface can substantially promote cell adhesion and proliferation. X‐ray photoelectron spectroscopy (XPS) has been used to characterise poly‐ε‐caprolactone (PCL) films modified by aminolysis and the introduction of RGD peptides via carbodiimide (CDI) and thiol‐halogen ‘click’ chemistry. The nitrogen signal acts as an elemental indicator for successful attachment, and changes in the chemical environment are reflected in the carbon and oxygen spectra. Chlorine and sulfur provide additional chemical indicators of reaction progress in the thiol method, with the selective nature of the Cl–S reaction reflected in the complete loss of Cl signal and appearance of S, avoiding potential amine‐peptide side‐reactions. Comparison of the XPS elemental concentrations indicated an estimate of 2–3% peptide functionalisation on the PCL surface for both methods, correlating with the improvement in Schwann cell response observed after peptide immobilisation. The enhanced selectivity of peptide attachment to the polymer surface demonstrated with XPS for the novel method based on thiol chemistry shows its potential for development as a biomimetic scaffold for peripheral nerve injury. Copyright © 2014 John Wiley &amp; Sons, Ltd.

Magnetic Study of the Heated and Unheated Sedimentary Fillings of Sebkha Mhabeul, Southeast Tunisia: A Geophysical Method for Paleoclimatic Investigation and Tephrochronological Dating

This paper is meant to investigate the climatic and volcanic signals within the sedimentary filling of sebkha Mhabeul through a thermomagnetic study of a 37 cm length core. Values of the magnetic susceptibility at ambient temperature show that the core encompasses four climatic stages: the Warming Present (WP), the Little Ice Age (Late LIA), Early Little Ice Age (ELIA), and the Medieval Climate Anomalies (MCA). Added to the subcycles, the spectral analysis shows the individualization of an 888 yr cycle probably related to solar activity. The heating at 250°C is good-for-nothing since it was useful neither for climatic investigation nor for tephras layers detection. Heating at 700°C generated the complete loss of the climatic signal. On the other hand, it allowed the detection of the previously identified tephras layers. Further, it highlighted the presence of other tephras layers. The extraction by the bromoform confirms the presence of these tephras. The use of the same methodology may allow the detection of tephras layers within other sebkhas.

A fuzzy extension of VIKOR for target network selection in heterogeneous wireless environments

In a highly integrated ubiquitous wireless environment, the selection of a network that can fulfill end-users’ service requests while keeping their overall satisfaction at a high level, is vital. The wrong selection can lead to undesirable conditions such as unsatisfied users, weak Quality of Service (QoS), network congestions, dropped and/or blocked calls, and wastage of valuable network resources. The selection of these networks is performed during the handoff process when a Mobile Station (MS) switches its current Point of Attachment (PoA) to a different network due to the degradation or complete loss of signal and/or deterioration of the provided QoS. Traditional schemes perform the handoff necessity estimation and trigger the network selection process based on a single metric such as Received Signal Strength (RSS). These schemes are not efficient enough, as they do not take into consideration the traffic characteristics, user preferences, network conditions and other important system metrics. This paper presents a novel multi-attribute vertical handoff algorithm for heterogeneous wireless networks which achieves seamless mobility while maximizing end-users’ satisfaction. Two modules are designed to estimated the necessity of handoff and to select the target network. These modules utilize parallel Fuzzy Logic Controllers (FLCs) with reduced rule-set in combination with a network ranking algorithm developed based on Fuzzy VIKOR (FVIKOR). Simulation results are provided and compared with a benchmark.

Micro-magnetic simulation study on the magnetic particle imaging performance of anisotropic mono-domain particles

The performance of magnetic mono-domain particles is of crucial importance in magnetic particle imaging (MPI). So far, the behavior of mono-domain particles has been modeled within the framework of Langevin theory. This theory predicts the dependence of the MPI signal on the particle core size, but cannot account for the influence of the shape, i.e. the anisotropy of the particle core. In this study we present the first micro-magnetic ab initio simulation of spectra of anisotropic particles with different core diameters in an oscillating magnetic field at 25 and 100 kHz. We find that the MPI signal strongly depends on the anisotropy of the magnetic core. Thus, a difference of 3 nm between the principal axes of a prolate ellipsoid with the volume of a 30 nm sphere can result in a complete loss of the MPI signal. Smaller anisotropies, however, can increase the MPI performance of the particle. The simulations show that the effect of the anisotropy on the MPI signal depends on the frequency of the oscillating magnetic field. At 100 kHz, the optimal signal is found at smaller anisotropies than at 25 kHz. Furthermore, the simulations show that experimental spectroscopic results for Resovist® can only be explained quantitatively by particles with a magnetic core size of at least 25 nm.