Processing of the circumsporozoite protein in infected hepatocytes is not dependent on aspartic proteases

CD8⁺ T cells play a major role in the protective immune response against the liver stage of malaria. It was previously shown that the circumsporozoite protein (CSP) is processed and presented to specific T cells by both traversed and infected hepatocytes, but their respective antigen processing requirements were not completely defined. In the present study, we show that in vitro processing of the Plasmodium berghei CSP by infected mouse primary hepatocytes is exclusively dependent on proteasomes, while aspartic proteases are also needed in the case of traversed hepatocytes.     

Ecodesign of the Aluminum Bronze Cutting Process

The realization of products from materials with high properties generally involves very high energy consumption. Thus, in the research, it was considered to optimize the machining process by cutting of an aluminum bronze alloy, so as to obtain a reduction in energy consumption in correlation with the roughness of the machined surfaces. The research focused on the processing of a semi-finished product with a diameter of Ø = 20 mm made of aluminum bronze ({"type":"entrez-nucleotide","attrs":{"text":"C62300","term_id":"56147584","term_text":"C62300"}}C62300). In addition, in the research, the aim was to establish some correlations between the amount of power consumed and the quality of the surfaces processed by cutting. In this sense, the forces were measured in the 3 directions specific to the cutting process (F_c; F_f; F_p) for 3 tools construction variants and power consumed. The results showed that, if a certain constructive variant of the cutting tool is used in the processing, a reduction of the power consumed to cutting can be obtained by approximately 30% and a reduction of the roughness of the processed surface by approximately 90–100%. Furthermore, following the statistical processing of the results, it was shown that it would be advisable to use, especially in roughing processes, the cutting tool variant that offers the greatest reduction in roughness and cutting power.     

Study on Platinum Coating Depth in Focused Ion Beam Diamond Cutting Tool Milling and Methods for Removing Platinum Layer

In recent years, nanomachining has attracted increasing attention in advanced manufacturing science and technologies as a value-added processes to control material structures, components, devices, and nanoscale systems. To make sub-micro patterns on these products, micro/nanoscale single-crystal diamond cutting tools are essential. Popular non-contact methods for the macro/micro processing of diamond composites are pulsed laser ablation (PLA) and electric discharge machining (EDM). However, for manufacturing nanoscale diamond tools, these machining methods are not appropriate. Despite diamond’s extreme physical properties, diamond can be micro/nano machined relatively easily using a focused ion beam (FIB) technique. In the FIB milling process, the surface properties of the diamond cutting tool is affected by the amorphous damage layer caused by the FIB gallium ion collision and implantation and these influence the diamond cutting tool edge sharpness and increase the processing procedures. To protect the diamond substrate, a protection layer—platinum (Pt) coating is essential in diamond FIB milling. In this study, the depth of Pt coating layer which could decrease process-induced damage during FIB fabrication is investigated, along with methods for removing the Pt coating layer on diamond tools. The optimum Pt coating depth has been confirmed, which is very important for maintaining cutting tool edge sharpness and decreasing processing procedures. The ultra-precision grinding method and etching with aqua regia method have been investigated for removing the Pt coating layer. Experimental results show that when the diamond cutting tool width is bigger than 500 nm, ultra-precision grinding method is appropriate for removing Pt coating layer on diamond tool. However, the ultra-precision grinding method is not recommended for removing the Pt coating layer when the cutting tool width is smaller than 500 nm, because the possibility that the diamond cutting tool is damaged by the grinding process will be increased. Despite the etching method requiring more procedures to remove the Pt coating layer after FIB milling, it is a feasible method for diamond tools with under 500 nm width.     

Leukapheresis for the extraction of monocytes and various lymphocyte subpopulations from peripheral blood: product quality and prediction of the yield using different harvest procedures

BACKGROUND AND OBJECTIVES: Leukapheresis of non-mobilized healthy donors is performed to harvest monocytes and lymphocyte subpopulations for use in various therapeutic regimens. In this methodological study, we compared two different leukapheresis programs, using equivalent volumes of processed blood over similar processing periods, to determine the influence of the procedures on the donor peripheral blood count and to establish the procedure that yields the highest quality product. MATERIALS AND METHODS: The target variables obtained in 41 healthy blood donors who underwent short-term leukapheresis (80-105 min) were retrospectively compared. Twenty-one volunteers were processed on a COBE Spectra machine at the MNC setting and 20 volunteers were processed at the AutoPBSC setting. Data were collected on pre- and postleukapheresis samples and on the product. RESULTS: AutoPBSC and MNC procedures resulted in a decrease of haemoglobin (5-7%), platelets (17-20%), monocytes (22%) and lymphocytes (23-27%), but not of granulocytes in peripheral blood. Both procedures produced nearly identical leucocyte and lymphocyte yields. AutoPBSC products contained a greater number of granulocytes, monocytes and red cells, but fewer platelets. The preleukapheresis values correlated with the yields for monocytes, T-helper and T-suppressor cells, B-lymphocytes and natural killer cells, but not for granulocytes or platelets. CONCLUSIONS: Leukapheresis is a safe and efficient procedure for collecting large numbers of peripheral blood monocytes and different lymphocyte populations from non-mobilized donors. The two programs yield comparable leucocyte harvests. Based on our results, yields can be predicted from the peripheral cell counts.     

Compensatory network changes in the dentate gyrus restore long-term potentiation following ablation of neurogenesis in young-adult mice

It is now well established that neurogenesis in the rodent subgranular zone of the hippocampal dentate gyrus continues throughout adulthood. Neuroblasts born in the dentate subgranular zone migrate into the granule cell layer, where they differentiate into neurons known as dentate granule cells. Suppression of neurogenesis by irradiation or genetic ablation has been shown to disrupt synaptic plasticity in the dentate gyrus and impair some forms of hippocampus-dependent learning and memory. Using a recently developed transgenic mouse model for suppressing neurogenesis, we sought to determine the long-term impact of ablating neurogenesis on synaptic plasticity in young-adult mice. Consistent with previous reports, we found that ablation of neurogenesis resulted in significant deficits in dentate gyrus long-term potentiation (LTP) when examined at a time proximal to the ablation. However, the observed deficits in LTP were not permanent. LTP in the dentate gyrus was restored within 6 wk and this recovery occurred in the complete absence of neurogenesis. The recovery in LTP was accompanied by prominent changes within the dentate gyrus, including an increase in the survival rate of newborn cells that were proliferating just before the ablation and a reduction in inhibitory input to the granule cells of the dentate gyrus. These findings suggest that prolonged suppression of neurogenesis in young-adult mice results in wide-ranging compensatory changes in the structure and dynamics of the dentate gyrus that function to restore plasticity.     

Catheter ablation of ventricular tachycardia by intramyocardial injection of ethanol in an animal model of chronic myocardial infarction

INTRODUCTION: Direct injection of ethanol into myocardium has been shown to create large, well-demarcated lesions with transmural necrosis in normal ventricular myocardium and in regions of healed myocardial infarction. The aim of this study was to investigate the effects of direct ethanol injection on the inducibility of ventricular tachycardia (VT) in an animal model of chronic myocardial infarction. METHODS AND RESULTS: Eight sheep with reproducibly inducible VT underwent an electrophysiologic study 139 +/- 65 days after myocardial infarction. Noncontact mapping was used to analyze induced VT. Fifteen different VTs were targeted for catheter ablation. Ablation was achieved by catheter-based intramyocardial injection of a mixture of 96% ethanol, glycerine, and iopromide (ratio 3:1:1). Direct intramyocardial ethanol injection resulted in noninducibility of any VT 20 minutes after ablation in 7 of 8 animals. Four of 5 animals with initially successful ablation remained noninducible for any VT at follow-up study at least 2 days after the ablation procedure. Microscopic examination revealed homogeneous lesions with interstitial edema, intramural hemorrhage, and myofibrillar degeneration at the lesion border. The lesions were well demarcated from the surrounding tissue by a border zone of neutrophilic infiltration. CONCLUSION: Catheter ablation of VT by direct intramyocardial injection of ethanol during the chronic phase of myocardial infarction is feasible. It may be a useful tool for catheter ablation when the area of interest is located deep intramyocardially or subepicardially or when a more regional approach requires ablation of larger amounts of tissue.     

Propidium iodide compares favorably with histology and triphenyl tetrazolium chloride in the assessment of experimentally-induced infarct size

Quantitative assessment of infarct size after histology processing or by tetrazolium staining requires long reperfusion times to facilitate the delineation of the injured tissue. We evaluated the nucleic acid stain propidium iodide as an alternate technique for assessment of infarct size because it does not require extended reperfusion and is accomplished with more simple tissue processing. Eight mice underwent 45 min of coronary artery occlusion and 24 h of reperfusion, after which propidium iodide was administered and allowed to circulate for 15 min. Hearts were excised, sliced into transverse slices approximately 620 micro m thick, and photographed for determination of infarction with propidium iodide. Slices were processed by standard histology techniques, and infarction was evaluated using classical criteria of necrosis. In four other mice, infarct size was assessed by both triphenyl tetrazolium chloride and propidium iodide staining. A total of 46 slices were analysed and infarction was described as a percentage of the cross-sectional area of the slice. The correlation (r=0.94), orthogonal regression line (y=0.9x+1.8), and the percentage of the cross-section infarcted (histology 17.5+/-3.0%v propidium iodide 18.5+/-3.0%), demonstrates that infarct size assessment after propidium iodide staining or histology processing yields the same results. Similarly, propidium iodide measurement of infarct size was comparable to that obtained with TCC staining (22.5+/-4.1%v 25.4+/-5.4%, respectively, r=0.84). We conclude that propidium iodide staining yields accurate infarct size assessment and is a simpler alternative to tetrazolium staining or histologic processing.     

Post-translational modification of Fibroblast Growth Factor 23

Fibroblast Growth Factor (FGF) 23 has been shown to play important roles in the development of hypophosphatemic rickets/osteomalacia. Complementary DNA predicts that the FGF23 protein is composed of 251 amino acids and N-terminal 24 amino acids seem to be a signal peptide. In vitro experiments indicate that a part of the FGF23 protein is processed between arginine179 and serine180. When full-length, N-terminal and C-terminal processed fragments of FGF23 were injected into mice, only the full-length FGF23 reduced serum phosphate levels indicating that the processing of FGF23 abolished its effect to cause hypophosphatemia. This processing was shown to be prevented by an inhibitor of furin indicating that the cleavage is mediated by subtilisin-like proprotein convertase. In addition to this processing, FGF23 protein seems to have O-linked glycosylation. Further studies are necessary to clarify the importance of O-glycosylation for FGF23 activity.     

Age differences in the effects of perceptual noise.

Elderly (60 to 82 years) and young (18 to 25 years) subjects were tested on a visual task in which noise items were present but did not require processing to test the hypotheses that elderly persons have more difficulty ignoring irrelevent stimuli and suppressing response competition than do younger persons. Tachistoscopically presented displays contained a precued central target letter either alone or flanked by noise items that required (1) a response the same as, or (2) opposite to that required by the target letter, or (3) did not require a response in this task. No evidence was found to support differentially greater response slowing by the elderly due to the presence of irrelevant stimuli. Elderly persons showed significantly less effects of response competition, suggesting that the two groups processed the task information differently. The results suggest that age differences in effects of perceptual noise and response competition are not general phenomena, but depend upon the demand characteristics of the specific task.     

Structural variety of arginine-rich RNA-binding peptides.

Arginine-rich domains are used by a variety of RNA-binding proteins to recognize specific RNA hairpins. It has been shown previously that a 17-aa arginine-rich peptide from the human immunodeficiency virus Rev protein binds specifically to its RNA site when the peptide is in an alpha-helical conformation. Here we show that related peptides from splicing factors, viral coat proteins, and bacteriophage antiterminators (the N proteins) also have propensities to form alpha-helices and that the N peptides require helical conformations to bind to their cognate RNAs. In contrast, introducing proline mutations into the arginine-rich domain of the human immunodeficiency virus Tat protein abolishes its potential to form an alpha-helix but does not affect RNA-binding affinity in vitro or in vivo. Based on results from several peptide-RNA model systems, we suggest that helical peptides may be used to recognize RNA structures having particularly wide major grooves, such as those found near loops or large bulges, and that nonhelical or extended peptides may be used to recognize less accessible grooves.     

On the Ablation Behavior of Carbon Fiber-Reinforced Plastics during Laser Surface Treatment Using Pulsed Lasers

This contribution discusses the ablation phenomena observed during laser treatment of carbon fiber-reinforced plastics (CFRPs) with pulsed lasers observed employing laser sources with wavelengths of 355 nm, 1064 nm and 10.6 µm and pulse durations from picoseconds (11 ps) to microseconds (14 µs) are analyzed and discussed. In particular, the threshold fluence of the matrix material epoxy (EP) and the damage threshold of CFRP were calculated. Moreover, two general surface pretreatment strategies are investigated, including selective matrix removal and structure generation through indentation (ablation of both, matrix material and fibers) with a cross-like morphology. The surfaces obtained after the laser treatment are characterized by means of optical and scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy is employed for the analysis of composite and constituent materials epoxy and carbon fibers. As a result, different ablation mechanisms, including evaporation and delamination are observed, depending on the employed laser wavelength and pulse duration. For both 355 nm and 1064 nm wavelength, the laser radiation produces only partial ablation of the carbon fibers due to their higher absorption coefficient compared to the epoxy matrix. Although a selective matrix removal without residues is achieved using the pulsed CO₂ laser. Differently, both constituent materials are ablated with the nanosecond pulsed UV laser, producing indentations. The sum of the investigations has shown that existing theories of laser technology, such as the ablation threshold according to Liu et al., can be applied to composite materials only to a limited extent. Furthermore, it has been found that the pronounced heterogeneity of CFRP mostly leads to an inhomogeneous ablation result, both when creating grooves and during selective matrix removal, where the carbon fibers influence the ablation result by their thermal conductivity, depending on fiber direction. Finally, despite the material inhomogeneity, a scanning strategy has been developed to compensate the heterogeneous ablation results regarding structure depth, width and heat affected zone.     

CHO cells synthesize amidated neuropeptide Y from a C-peptide deleted form of the precursor

Post-translational processing of peptide precursors producing amidated, biologically active peptides is generally believed to occur only in specially differentiated endocrine or neural cells. Previously it has been shown that endoproteolytic processing of peptide precursors is very inefficient in non-endocrine cells like CHO cells. We have studied the processing of a C-peptide-deleted precursor of neuropeptide Y (NPY) in which the precursor terminates in the sequence Gly-Lys-Arg and does not require any dibasic specific endoproteolytic processing. Following transfection of CHO cells with an expression plasmid encoding this mutated NPY precursor, between 50 and 80 percent of the synthesized NPY was secreted from stable transfectants as authentic amidated NPY as assessed by both a C-terminal amide specific radioimmunoassay and by isoelectric focusing. It is concluded that amidated peptides can be produced in non-endocrine cells provided they are presented with a precursor which does not have to be endoproteolytically processed.     

Factors producing error in the total leucocyte count as measured on the Coulter Counter S Plus IVD

Summary Whole blood specimens, anticoagulated with potassium EDTA, which were seen to have particles interfering with the total leucocyte count were collected. This interference was detected, when the samples were processed through a Coulter Counter S Plus IVD, by R1 regional flags or backlighting of the leucocyte count. These specimens then had visual leucocyte counts performed using a counting chamber method and were examined microscopically, to ascertain possible causes of the interference. It was shown that the degree of error in the leucocyte count produced varies considerably but can be very high. Particles causing the error were found to include fibrin strands, platelet clumps, nucleated red cells and giant platelets.     

Human immunodeficiency virus protease expressed in Escherichia coli exhibits autoprocessing and specific maturation of the gag precursor.

The mature gag and pol proteins of human immunodeficiency virus (HIV) and all retroviruses derive from large gag and gag-pol polyprotein precursors by posttranslational cleavage. A highly specific, virally encoded protease is required for this essential proteolytic processing. In this study, the HIV protease gene product was expressed in Escherichia coli and shown to autocatalyze its maturation from a larger precursor. In addition, this bacterially produced HIV protease specifically processed an HIV p55 gag polyprotein precursor when coexpressed in E. coli. This system will allow detailed structure-function analysis of the HIV protease and provides a simple assay for the development of potential therapeutic agents directed against this critical viral enzyme.     

Real-time image equalization for coronary X-ray angiography.

Coronary angiograms, which provide detailed images of contrast-filled coronary arteries, also show other large structures such as the diaphragm, spine and adjacent lung field. A real-time image processing method to attenuate these unwanted features is presented. Side-by-side comparisons of images selected from cine runs before and after processing show that the arteries in the processed images can be visualized more easily due to their higher contrast as other structures are made less prominent. It is also shown experimentally that this method allows more quantitative comparisons of the contrast of vessels in different parts of an image.     

心房起搏法在射频消融房室结折返性心动过速中的应用

将射频消融治疗的９４例房室结折返性心动过速（ＡＶＮＲＴ）病人按心房起搏法和常规法进行分组（分别为３９及５５例），回顾性比较两组病人的消融治疗结果，以评价这两种方法在射频消融治疗ＡＶＮＲＴ中的安全性、成功率和复发率。随访１０．８±４．５个月，总成功率为９６．８％、复发率为２．１％。与常规组相比，起搏组有效放电时间明显延长（１４５±３８ｓｖｓ８２±２６ｓ，Ｐ＜０．０１）、慢径阻断成功率高（６１．５％ｖｓ４０．０％，Ｐ＜０．０１）、一过性房室阻滞发生率低（２．６％ｖｓ１２．７％，Ｐ＜０．０５），但各种类型的永久性房室阻滞发生率和复发率无显著性差异（Ｐ＞０．０５）。表明ＡＶＮ－ＲＴ消融术中采用心房起搏法较常规法更为安全有效。     

Experimental Analysis of Ductile Cutting Regime in Face Milling of Sintered Silicon Carbide

In this study, sintered silicon carbide is machined on a high-precision milling machine with a high-speed spindle, closed-loop linear drives and friction-free micro gap hydrostatics. A series of experiments was undertaken varying the relevant process parameters such as feedrate, cutting speed and chip thickness. For this, the milled surfaces are characterized in a process via an acoustic emission sensor. The milled surfaces were analyzed via confocal laser scanning microscopy and the ISO 25178 areal surface quality parameters such as Sa, Sq and Smr are determined. Moreover, scanning electron microscopy was used to qualitatively characterize the surfaces, but also to identify sub-surface damages such as grooves, breakouts and pitting. Raman laser spectroscopy is used to identify possible amorphization and changes to crystal structure. We used grazing incidence XRD to analyze the crystallographic structure and scanning acoustic microscopy to analyze sub-surface damages. A polycrystalline diamond tool was able to produce superior surfaces compared to diamond grinding with an areal surface roughness Sa of below 100 nm in a very competitive time frame. The finished surface exhibits a high gloss and reflectance. It can be seen that chip thickness and cutting speed have a major influence on the resulting surface quality. The undamaged surface in combination with a small median chip thickness is indicative of a ductile cutting regime.     

Radiofrequency catheter ablation of left ventricular tachycardia in the normal heart

Background: Radiofrequency (RF) catheter ablation is a safe and effective cure for many forms of supraventricular tachycardia. Its efficacy in the cure of right ventricular outflow tract tachycardia, and some forms of left ventricular tachycardia in patients with left ventricular dysfunction, has also been shown. In contrast limited data are available to assess the role of RF catheter ablation in treating idiopathic left ventricular tachycardia (ILVT), an unusual form of tachycardia occurring in patients without demonstrable heart disease.
Aim: To examine the efficacy and safety of RF catheter ablation in patients with ILVT.
Methods: Three patients without structural heart disease and with recurrent drug-refractory ILVT (right bundle branch block and left axis morphology) underwent electrophysiologic study (EPS) to initiate and localise the site of origin of their VT. RF catheter ablation of the VT focus was performed, with success being defined as failure to reinduce VT during incremental infusion of isoprenaline.
Results: In all three patients VT was inducible by rapid right atrial pacing and/or programmed ventricular stimulation, and could be terminated by intravenous verapamil. RF catheter ablation was successful in all patients. The site of successful ablation was common to each patient and was localised to the infero-apical aspect of the left ventricular septum. It was characterised by the recording of the earliest presystolic 'P' potential during both sinus rhythm and induced ILVT. No complications occurred during the procedure. During follow-up periods ranging from six to 12 months there were no symptomatic or documented episodes of recurrent ILVT.
Conclusions: We conclude that ILVT can be safely and effectively cured by RF catheter ablation.     

Energy-dependent processing of cytoplasmically made precursors to mitochondrial proteins.

Earlier work has shown that mitochondrial proteins synthesized in the cytosol are initially made as larger precursors which are then transferred into the organelles and processed to their mature size in the absence of protein synthesis. It is now demonstrated that depletion of the mitochondrial matrix ATP in intact yeast spheroplasts by various combinations of inhibitors and mutations prevents the processing of precursors to the three largest subunits of the mitochondrial F1-ATPase and two subunits of the cytochrome bc1 complex. These polypeptides are all synthesized outside the mitochondria and transported to the mitochondrial matrix or inserted into the mitochondrial inner membrane. In contrast, depletion of the matrix ATP does not inhibit processing of the precursor to cytochrome c peroxidase; this enzyme is located in the mitochondrial intermembrane space which is freely accessible to ATP made in the cytosol. The processing of extramitochondrially made precursors or the transfer of these precursors across the mitochondrial inner membrane is thus dependent on ATP.     

Improved Laser Ablation Method for the Production of Luminescent Carbon Particles in Liquids

An improved method for the production of luminescent carbon nanoparticles is proposed in this work. The new method overcomes the disadvantages of commonly used approaches. It involves two-stage laser ablation in water and in aqueous solutions, where the first stage is the laser ablation of a graphite target and the second is the shredding of particles produced in the first step. The two-stage method offers the optimization of the laser pulse fluence for the performance of each process. It was found that the two-stage process of laser ablation allows producing photoluminescent carbon structures in pure water. The additional reagent may be added either in the first or second stage. The first stage performed in pure water allows avoiding the contamination of the target. Moreover, it simplifies the identification of the origin of photoluminescence. Two synthesis routes for the preparation of carbon nanoparticles by the proposed method using pure water as well as urea aqueous solution are investigated. It was found that the use of urea as a reagent results in luminescence properties similar to those obtained with other more hazardous amine-based reagents. The influence of the synthesis approach and process parameters on the structural and luminescent properties of nanoparticles is also explored in this work.