Acute vasoconstriction after subarachnoid hemorrhage

OBJECTIVE: Decreased cerebral blood flow (CBF) and cerebral ischemia occurring immediately after subarachnoid hemorrhage (SAH) may be caused by acute microvascular constriction. However, CBF can also be influenced by changes in intracranial pressure (ICP) and cerebral perfusion pressure (CPP). The goal of these experiments was to assess the significance of acute vasoconstriction after SAH and its relationship to changes in CBF, ICP, CPP, and extracellular glutamate concentrations. METHODS: Three experiments were performed using the endovascular filament technique to produce SAH. In the first experiment, CBF, ICP, and CPP were measured for 60 minutes after SAH (n = 21) and were correlated with the 24-hour mortality rate. In the second experiment, rats undergoing SAH (n = 23) or a sham procedure (n = 7) were perfused 60 minutes after SAH for measurement of the circumference and wall thickness of the internal carotid and anterior cerebral arteries and correlation with CBF, ICP, and CPP. In the third experiment (n = 11), extracellular glutamate concentrations determined by hippocampal and cortical microdialysis and high performance liquid chromatography were correlated with physiological changes. RESULTS: CBF reductions to less than 40% of baseline for 60 minutes after SAH predicted 24-hour mortality with 100% accuracy and were used to define "lethal" SAH. In contrast, ICP and CPP 60 minutes after SAH were not correlated with the mortality rate. The vascular circumference was significantly smaller in lethal than in sublethal SAH or sham-operated rats (P < 0.001). Vessel measurements were correlated with both CBF and hemorrhage size (P < 0.01). Extracellular glutamate concentration increased to 600% of baseline after lethal SAH in both hippocampus and cortex and was inversely correlated with CBF (r = 0.9, P < 0.001) but did not increase after sublethal SAH. CONCLUSION: Acute vasoconstriction after SAH occurs independently of changes in ICP and CPP and is associated with decreased CBF, larger hemorrhage size, persistent elevations of extracellular glutamate, and poor outcome. Acute vasoconstriction seems to contribute directly to ischemic brain injury after SAH. Further evaluations of pharmacological agents with the potential to reverse acute vasoconstriction may increase CBF and improve outcome.     

The evaluation of lyophilized polymer matrices for administering recombinant human bone morphogenetic protein-2

Novel unitary devices, prepared by lyophilization of viscous solutions of sodium carboxymethylcellulose (CMC) and methylcellulose (MC), were evaluated as sustained-release delivery systems for recombinant human bone morphogenetic protein-2 (rhBMP-2). In vitro characterization of the unitary devices, which contained rhBMP-2-loaded poly (d,l lactide-co-glycolide) (PLGA) bioerodible particles (BEPs), was conducted over a 2-month period. Determinations included buffer uptake, mass and molecular weight loss and rhBMP-2 release from the unitary devices. CMC devices imbibed approximately 16 times their weight of buffer, while with MC, equilibrium uptake was approximately 6 times the dry weight of the devices. Overall mass loss percentages were approximately 55 and 35%, respectively, for CMC and MC devices. rhBMP-2 release from the devices was essentially a triphasic process: an initial phase during which "free" protein (rhBMP-2 present on the surface and within the pores of the PLGA BEPs) was released, a lag period during which no release was discerned, and then release of "bound" rhBMP-2 (protein adsorbed to the BEPs). The release of bound protein correlated with the mass loss of the polymer which began after 3 weeks. Release from the unitary devices was lower than that from the BEPs alone, due to a retardation effect of the gelled CMC/MC polymers. In rabbits in which full-thickness cranial bone defects were created, the implants were well tolerated and induced significant new bone growth during an 8-week evaluation period. The CMC devices appear to have induced bone earlier (at 2 weeks), but this did not affect eventual 8-week results. CMC devices without rhBMP-2 appeared to provide some bone conduction, in contrast to the blank MC devices.     

Sensitivity of the retinal pigment epithelium to spindle poisons

PURPOSE: The function of RPE is well known in PVR. Pharmacological agents have been extensively studied both experimentally and clinically. Few reports have detailed the interactions of antimitotic drugs on the microtubule network. The aim of this study is to visualize by indirect immunofluorescence the effects of colchicine and paclitaxel on the microtubule network of cultured pig RPE cells in interphase. METHODS: Pigs were killed at the slaughter-house, their eyes were enucleated. RPE cells were isolated and cultured. RPE cells were plated onto glass cover-slips at a density of 2,000,000 cells/ml, cultured and treated with the drugs during 4 and 24 hours at 37 degrees C at different concentrations. Immunofluorescence reaction was developped using antitubulin and fluoresceinated anti-mouse antibodies. The cytoskeletons were visualized employing a Zeiss photomicroscope equipped with epiilumination, a 63 x lens and appropriate filters for fluoresceine. RESULTS: The cytoplasmic microtubules of RPE cells were disrupted in a concentration and time-dependant manner by colchicine. Between 10 and 100 nm Veveral degrees of depolymarization of the microtubule network were observed. Paclitaxel between 1 micron and 10 microns was found to induce several degrees of microtubule "bundling" after 4 and 24 hours of incubation. Actin network was modified neither by colchicine and paclitaxel used in the same conditions. CONCLUSIONS: The results show that low doses of antimitotic drugs inhibit the microtubule network formation by depolymerization (colchicine) or stabilize it (paclitaxel). These actions inhibit cell division, which is one of the mechanisms implicated in PVR.     

Cell surface chondroitin sulfate proteoglycans in tumor cell adhesion, motility and invasion

Genes that encode enzymes that convert inactive "prodrugs" into anticancer metabolites may be therapeutically useful against brain tumors. Unlike other genes tested to date in brain tumor models, the Escherichia coli gpt gene is unique in that it not only sensitizes cells to the prodrug 6-thioxanthine (6TX) but also encodes resistance to a different regimen (mycophenolic acid, xanthine, and hypoxanthine), thus providing a means to select for gpt-positive cells. In the present study, rat C6 glioma cells were infected with a retrovirus vector that transduces this gene. A clonal line (C6GPT-7) was derived that exhibited significant 6TX susceptibility in vitro with an ID50 of 2.5 mumol/L, whereas 50% growth inhibition of parental C6 cells was not achieved at concentrations tested (up to 50 mumol/L). This line also exhibited significant sensitivity to 6-thioguanine (6TG), with an ID50 of 0.05 mumol/L, whereas 50% growth inhibition of parental C6 cells was achieved at 0.5 mumol/L. In a "bystander" assay, C6GPT-7 tumor cells efficiently transferred 6TX sensitivity to C6 cells at ratios as low as 1:9 (C6GPT-7:C6). This in vitro bystander effect was abrogated when C6GPT-7 and C6 cells were separated by a microporous membrane, suggesting that it was not mediated by highly diffusible metabolites. In vivo both 6TX and 6TG significantly inhibited the growth of subcutaneously transplanted C6GPT-7 cells but not that of C6 cells in athymic mice. In an intracerebral model, both 6TX and 6TG exhibited significant antiproliferative effects against tumors formed by C6GPT-7 cells. These findings provide a basis for exploring further gene therapy strategies based on in vivo transfer of the E coli gpt gene to provide chemosensitivity against 6TX and 6TG.     

Piezoelectric cantilever sensors with asymmetric anchor exhibit picogram sensitivity in liquids

We show for the first time that self-exciting and self-sensing piezoelectric cantilevers consisting only of lead zirconate titanate (PZT) measure resonance only if they are asymmetrically anchored. Symmetric-anchoring did not give rise to electrically measurable bending resonant modes in the 0-100 kHz range. Sensitivity of first and second bending mode resonances was characterized in a flow apparatus using small density changes in liquid (0.003-0.01 g/cm³) and by dodecanethiol chemisorption at 30 pM. Density change results were consistent with existing models of submerged cantilevers, and yielded mass-change sensitivity of ∼33 ng/Hz and 217 pg/Hz for the first two modes. In chemisorption experiments, where binding was localized to 1 mm² distal tip of the PZT cantilever, sensitivity improved by an order of magnitude to 2 pg/Hz and 414 fg/Hz for the same two resonant modes.     

SVM based robust watermarking for enhanced medical image security

Medical images are more typical than any other ordinary images, since it stores patient’s information for diagnosis purpose. Such images need more security and confidentiality as total diagnosis depends on it. In telemedicine applications, transmission of medical image via open channel, demands strong security and copyright protection. In our proposed robust watermarking model, a double layer security is introduced to ensure the robustness of embedded data. The embedded data is scrambled using a unique key and then a transform domain based hybrid watermarking technique is used to embed the scrambled data into the transform coefficients of the host image. The data embedding in medical images involves more attention, so that the diagnosis part must not be affected by any modification. Therefore, Support Vector Machine (SVM) is used as a classifier, which classify a medical image into two regions i.e. Non Region of Interest (NROI) and Region of Interest (ROI) to embed watermark data into the NROI part of the medical image, using the proposed embedding algorithm. The objective of the proposed model is to avoid any quality degradation to the medical image. The simulation is performed to measure the Peak Signal to Noise Ratio (PSNR) for imperceptibility and Structural Similarity Index (SSIM) to test the robustness. The experimented result shows, robustness and imperceptibility with SSIM of more than 0.50 and PSNR of more than 35 dB for proposed watermarking model.

     

Simultaneous automated design of structured QFT controller and prefilter using nonlinear programming

This paper describes a nonlinear programming‐based robust design methodology for controllers and prefilters of a predefined structure for the linear time‐invariant systems involved in the quantitative feedback theory. This controller and prefilter synthesis problem is formulated as a single optimization problem with a given performance optimization objective and constraints enforcing stability and various specifications usually enforced in the quantitative feedback theory. The focus is set on providing constraints expression that can be used in standard nonlinear programming solvers. The nonlinear solver then computes in a single‐step controller and prefilter design parameters that satisfy the prescribed constraints and maximizes the performance optimization objective. The effectiveness of the proposed approach is demonstrated through a variety of difficult design cases like resonant plants, open‐loop unstable plants, and plants with variation in the time delay. Copyright © 2016 John Wiley & Sons, Ltd.     

What can the canonical controller in principle tell us?

Given a plant and a desired specification our goal is to construct a controller system which, when interconnected with the plant, yields a system that behaves like the desired specification. We can always construct the canonical controller introduced in van der Schaft (2003) [10]. For linear systems there exists a controller which when interconnected to the plant yields the desired behaviour if and only if the canonical controller is itself one such controller, see Vinjamoor and van der Schaft (2011) [4]. In this paper we extend this result to nonlinear systems. It turns out that one has to look at the canonical controller together with its subsystems. We obtain necessary and sufficient conditions for the existence of a controller for a class of nonlinear systems. We end with examples which show that in certain cases looking at subsystems of the canonical controller also does not suffice.     

Preparation and properties of mitochondria derived from synaptosomes

A method has been developed whereby a fraction of rat brain mitochondria (synaptic mitochondria) was isolated from synaptosomes. This brain mitochondrial fraction was compared with the fraction of "free" brain mitochondria (non-synaptic) isolated by the method of Clark & Nicklas (1970). (J. Biol. Chem. 245, 4724-4731). Both mitochondrial fractions are shown to be relatively pure, metabolically active and well coupled. 2. The oxidation of a number of substrates by synaptic and non-synaptic mitochondria was studied and compared. Of the substrates studied, pyruvate plus malate was oxidized most rapidly by both mitochondrial populations. However, the non-synaptic mitochondria oxidized glutamate plus malate almost twice as rapidly as the synaptic mitochondria. 3. The activities of certain tricarboxylic acid-cycle and related enzymes in synaptic and non-synaptic mitochondria were determined. Citrate synthase (EC 4.1.3.7), isocitrate dehydrogenase (EC 1.1.1.41) and malate dehydrogenase (EC 1.1.1.37) activities were similar in both fractions, but pyruvate dehydrogenase (EC 1.2.4.1) activity in non-synaptic mitochondria was higher than in synaptic mitochondria and glutamate dehydrogenase (EC 1.4.1.3) activity in non-synaptic mitochondria was lower than that in synaptic mitochondria. 4. Comparison of synaptic and non-synaptic mitochondria by rate-zonal separation confirmed the distinct identity of the two mitochondrial populations. The non-synaptic mitochondria had higher buoyant density and evidence was obtained to suggest that the synaptic mitochondria might be heterogeneous. 5. The results are also discussed in the light of the suggested connection between the heterogeneity of brain mitochondria and metabolic compartmentation.