Spectral-Element Method for Levy-Type Plates Subject to Dynamic Loads

Spectral-element method (SEM) is applied for the vibration analysis of the Levy-type rectangular plates subject to distributed dynamic loads. In the solution procedure, both FFT and inverse FFT computer algorithms are efficiently used to obtain the dynamic responses in time domain. Distributed dynamic load is approximated, as the superposition of equivalent dynamic line loads and the spatial coordinate dependence of equivalent line loads is removed by (spatial) FFT. This procedure transforms the original plate (two-dimensional) problem into an effective beam (one-dimensional) problem so that the solution procedure for one-dimensional structures can be used. Numerical tests show that SEM provides very accurate solutions when compared with conventional finite element solutions.     

Intestinal phospholipase, a novel enzyme

We evaluated phospholipase activity in the intestine of rats and other species. Phospholipase activity was assayed by a surface barostat technique or an egg yolk titration system. Mucosal activity was found only by the surface barostat technique with phosphatidylglycerol as substrate; it was not found with phosphatidylcholine as substrate in assays by either technique. In gut luminal fluid activity was found when both phosphatidylcholine and phosphatidylglycerol were used as substrate in assays by the surface barostat technique, and phosphatidylcholine as substrate yielded activity in egg yolk titration. In rats in which pancreatic juice had been diverted, mucosal and gut luminal phospholipase activity was greater than in controls, thus demonstrating that enzyme activity was not due to pancreatic phospholipase. Bacterial origin of phospholipase activity was excluded in that phospholipase activity was found in germ-free rats; gastric and salivary gland origins were excluded in that continued phospholipase activity was found in rats with gastric fistula. The physiological importance of the enzyme was established by the finding that rats with pancreatic fistula absorbed 111 mumol of phosphatidylcholine and that controls absorbed 119 mumol of a 135-mumol load. Activity was found to be three times greater in the distal than in the proximal intestine; in cryptal cells it was 10 times greater than in villus tip cells. 65% of the activity in the gut lumen was tightly bound to particulate matter. We propose that intestinal phospholipase may be important in gut bacterial control, in the digestion of vegetable matter (phosphatidylglycerol is a major phospholipid in both plants and bacteria), and in the digestion of phospholipids in the gut lumen.     

Digital quantification of the enzyme-linked immunospot (ELISPOT)

Enzyme-linked immunospot (ELISPOT) is a sensitive technique for the detection of cytokines released by immune cells. The technique is well established and correlates closely with the enzyme-linked immunosorbent assay (ELISA) technique. Here, we introduce an integrated imaging-analysis system to quantitate the spots formed by the ELISPOT assay. The spots can be easily counted when the background is clear and the spots are few. However, when the number of spots increases to > 100, this task becomes challenging and time-consuming. To minimize time and standardize the counting procedure, we have used a digital camera linked to a computer system for reading the plates. In general, the computer is able to count more spots and has a smaller standard deviation when compared with the manual microscopic count. This integrated system is commercially available, and we believe that this method is objective, time-saving and consistent in routine ELISPOT assay counting, especially when numerous spots are present.     

Hidden anti-phospholipid antibodies in normal human sera circulate as immune complexes whose antigen can be removed by heat, acid, hypermolar buffers or phospholipase treatments

Heat treatment of normal human serum reveals otherwise masked anti-cardiolipin antibodies (aCL). We studied the mechanism of masking and the nature of the inhibitor of these aCL IgG. Other forms of treatment, besides heating for 30 min at 56 degrees C, can also unmask hidden aCL IgG. These include acid pH, hypermolar buffers and phospholipase digestion. When unmasked, these aCL recognize other anionic and zwitterionic phospholipids, but do not react with DNA, cell antigens or IgG. Using thin layer chromatography we demonstrate that the heat-labile inhibitor(s) of these aCL are phosphatidylserine, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine. These antibodies are not beta2-glycoprotein-I dependent and actually compete with this protein for phospholipid binding. The hidden antibodies are comprised of two populations of IgG autoantibodies: one reactive with cardiolipin, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol, phosphatidylethanolamine and sphingomyelin, and the other reactive almost exclusively with phosphatidylcholine and phosphorylcholine on enzyme-linked immunosorbent assay plates or when exposed by bromelain on the erythrocyte surface. Our data suggest that hidden aCL are natural oligoreactive IgG anti-phospholipid autoantibodies that circulate masked by their antigen.     

Hypotheses: melatonin/steroid combination contraceptives will prevent breast cancer

LDL, the major carrier of cholesterol in blood, is poorly metabolized by macrophages. In contrast, macrophages can recognize and endocytose anionic phospholipids such as phosphatidylserine, phosphatidylglycerol and cardiolipin. Since macrophages can take up large amounts of these phospholipids, experiments were performed to ascertain whether pre-incubation of native LDL with negatively-charged phospholipids would enhance the metabolism of LDL by macrophages. When 125I-LDL was incubated with cardiolipin liposomes for 18 h at 37 degrees C before addition to macrophages, an approx. 40-fold increase of LDL metabolism by these cells was observed. Similar results were found when LDL was pre-incubated with phosphatidylserine or phosphatidylglycerol; however, pre-incubation of LDL with phosphatidylcholine liposomes did not lead to an increase of LDL metabolism. The macrophage uptake of LDL pre-incubated with cardiolipin was reduced to approx. 40% of control values in the presence of dextran sulfate and fucoidin, inhibitors of anionic phospholipid uptake. Cytochalasin D, an inhibitor of phagocytosis, reduced the lysosomal degradation of LDL pre-incubated with cardiolipin to approx. 10% of control values. When the LDL-cardiolipin mixture was chromatographed on agarose gel, two peaks containing LDL were observed in the elution profile: the first peak appeared at the void volume and the second peak was detected just ahead of native LDL. The LDL in both peaks was much more extensively metabolized by macrophages than was native LDL; the LDL in the first peak was metabolized at a rate that was 8 times the second peak. The results demonstrate that negatively-charged phospholipids can form a complex with LDL which facilitates its phagocytosis by macrophages.     

Protein-lipid interactions and enzyme requirements for light subtype generation on cycling reconstituted surfactant

Surfactant convertase is required for conversion of heavy density (H) natural surfactant to light density (L) subtype during cycling in vitro, a technique that reproduces surfactant metabolism. To study mechanisms of H to L conversion, we prepared liposomes of dipalmitoylphosphatidylcholine (DPPC) and phosphatidylglycerol (PG), or the phospholipids (PL) in combination with either surfactant protein A (SP-A), surfactant protein B (SP-B), or both SP-A and SP-B. Phospholipids alone showed time-dependent conversion from heavy to light subtype on cycling in the absence of convertase, which was decreased by adding SP-B, but not SP-A, to phospholipids (p < 0.01 for PL+SP-B, or PL+SP-A+SP-B vs. PL, or PL+SP-A). The ultrastructure, surface activity, buoyant density, and L subtype generation on cycling PL+SP-A+SP-B with partially purified convertase or with phospholipase D were similar to those of natural TM. In conclusion, a reconstituted surfactant mimics the behavior of natural surfactant on cycling, and reveals that interaction of SP-B with phospholipids decreases L subtype generation. In addition, esterase/ phospholipase D activity is required for conversion of heavy to light subtype on cycling.     

Do patients with antiphospholipid syndrome have autoantibodies to beta 2-glycoprotein I?

High positive anticardiolipin antibody tests have been associated with recurrent thrombosis and pregnancy loss. Although these antibodies were believed to bind negatively charged phospholipids, recent reports have suggested that a serum protein, beta 2-glycoprotein I (beta 2-GPI), may be the true antigen for these antibodies. To resolve this issue, we compared binding of 75 anticardiolipin-positive and 71 anticardiolipin-negative serum samples from patients with rheumatic diseases to beta 2-GPI by using an enzyme-linked immunosorbent assay (ELISA). Serum samples from 30 healthy blood donors and 10 laboratory personnel were used as normal controls. We found no difference in binding between the three groups of serum samples. In addition, when binding to beta 2-GPI coated plates was compared with binding to ELISA plates without beta 2-GPI (blank), no difference was observed. Finally, binding of anticardiolipin-positive serum samples to plates coated with cardiolipin-beta 2-GPI mixture varied directly with the cardiolipin concentrations. Based on these findings, we conclude that anticardiolipin-positive serum samples do not bind beta 2-GPI.     

Extension of Preissmann Scheme to Two-Dimensional Flows

A numerical solution to the finite difference of two-dimensional (2D) depth-averaged equations on nonstaggered grid points is proposed in this technical note. Following a locally one-dimensional procedure, the basic equations are split into a pair of one-dimensional equations. Therefore, the solution of a 2D problem is reduced to the solution of a sequence of two one-dimensional problems. The discretization of the split one-dimensional equations is obtained with the use of the original Preissmann operator. Using Fourier’s classic linear analysis, stability, dissipation and dispersion with frictional resistance are investigated for the variations of the Courant number and weighting time factor.     

Echographic evaluation of glaucoma shunts

BACKGROUND: It is frequently impossible to determine by clinical examination alone if a filtering bleb is present over the equatorial plate(s) of a glaucoma shunt. This uncertainty can result in management dilemmas when patients with these devices have elevated intraocular pressures (IOPs). The authors describe how standardized echography can be used to determine if fluid is present around the plates and to characterize the blebs. METHODS: Four hundred twenty-seven plates in 269 patients who had undergone single- or double-plate Molteno implantation underwent standardized echography (both A- and B-scan examinations) to determine if blebs were present around the plates, primarily when results of slit-lamp examinations had been inconclusive. Bleb sizes, locations, and characteristics also were evaluated. RESULTS: Sixty-five percent of the plates had associated blebs, which were graded according to their sizes (small, medium, and large, in 29%, 44%, and 27% of plates with blebs, respectively). Fluid (which appeared as an echolucent area) could be seen underlying and/or overlying the plates (which appeared as echodense lines). Larger blebs were often associated with scleral flattening. Although bleb sizes did not necessarily correlate with the levels of IOP control, the presence of a bleb on ultrasound was indicative of tube patency. CONCLUSION: Standardized echography was invaluable in the postoperative management of patients who had undergone Molteno implantation because it demonstrated the presence or absence of blebs and characterized them when results of slit-lamp examinations had been inconclusive. This technique also can be used to evaluate similarly designed aqueous shunting devices.     

New Approach for Predicting Flow Bifurcation at Right-Angled Open-Channel Junction

An unsteady mathematical model for predicting flow divisions at a right-angled open-channel junction is presented. Existing dividing models depend on a prior knowledge of a constant flow regime. In addition, their strong nonlinearity does not guarantee compatibility with the St. Venant solutions in the context of an internal boundary condition treatment. Assuming zero crest height at the junction region, a side weir model explicitly introduced within the one-dimensional St. Venant equations is used to cope with the two-dimensional pattern of the flow. An upwind implicit numerical solver is employed to compute the new governing equations. The performance of the proposed technique in predicting super-, trans-, and subcritical flow bifurcations is illustrated by comparing with experimental data and/or theoretical predictions. In all the tests, lateral-to-upstream discharge ratios (Rq) are successfully reproduced by the present technique with a maximum error magnitude of less than 9%.     

Binding of quinacrine to acidic phospholipids and pancreatic phospholipase A2. Effects on the catalytic activity of the enzyme

Binding of quinacrine to phospholipids and porcine pancreatic phospholipase A2 (PLA2) was investigated using fluorescence resonance energy transfer, Langmuir films, assay for the enzymatic activity, and molecular modeling. No significant binding of this drug to the zwitterionic phosphatidylcholine was observed whereas a high affinity for acidic phospholipids was revealed by quenching of pyrene-labeled phospholipid analogues. Partial reversal of this binding was observed due to the addition of 4 mM CaCl2. Quinacrine efficiently and independently of the lipid surface pressure penetrated into monolayers of phosphatidylglycerol while only a weak penetration into phosphatidylcholine films was evident. Quinacrine also bound to eosin-labeled PLA2, and the addition of 4 mM CaCl2 reversed this interaction almost completely. In the presence of acidic phospholipids both the drug and the enzyme were attached to the lipid surface. Studies on the influence of quinacrine on the activity of PLA2 toward pyrene-labeled phospholipid analogues revealed that the hydrolysis of phosphatidylcholine was progressively reduced as a function of increasing [quinacrine]. At low [CaCl2] and low quinacrine:lipid molar ratios (<1:5) quinacrine enhanced slightly the rate of hydrolysis of acidic phospholipids whereas at higher drug:lipid molar ratios (>1:2) an inhibition was observed. In the presence of 1 mM CaCl2 quinacrine inhibited PLA2-catalyzed hydrolysis of phosphatidylglycerol only when the drug:lipid molar ratio exceeded 1:1. The presence of 4 mM CaCl2 abolished nearly completely the inhibition with all the substrate analogues used. Our data suggest that the inhibition of PLA2 by quinacrine is due to its binding to the enzyme. This is supported also by molecular modeling which suggested a binding site for quinacrine close to the active site and Ca2+ binding site of the enzyme. Importantly, our data indicate that quinacrine binds avidly to acidic phospholipids and their presence may influence the drug-enzyme interaction and the inhibition of the enzyme action. Accordingly, presence of quinacrine may interfere also with other processes that require the presence of acidic lipids and/or Ca2+, such as the function of the nicotinic acetylcholine receptor.     

Purification and properties of penicillin acylase of Bovista plumbea

Lipids from the insoluble material obtained by pulmonary lavage of 6 patients with alveolar proteinosis and from lamellar organelles of normal rabbit lungs were isolated and characterized. In both types of samples, dipalmitoylphosphatidylcholine was the predominant lipid. Phosphatidylethanolamine, phosphatidylglycerol, lysophosphatidylglycerol, and 2 glycolipids, GM3 and GL1 were also present in both types of preparations. Sphingomyelin, lysophosphatidylcholine, lysophosphatidylethanolamine, phosphatidyl-N, N-dimethylethanolamine, phosphatidylserine, and lyso(bis)phosphatidic acid were found in the sedimented lavage material from humans but were not detected in lamellar organelles from rabbits. Significant quantities of neutral lipids were present in the lavage material, but only trace amounts, mainly as cholesterol and triglycerides, were detected in lamellar organelles. Phosphatidylcholine and the 2 glycolipids contained mostly saturated fatty acids and essentially no polyunsaturated fatty acids. Sphingomyelin, lysophosphatidycholine, and phosphatidyl-N, N-dimethylethanolamine, found only in the lavage, were also highly saturated. In addition to the fact that several phospholipids found in the lavage were not present in lamellar organelles, another striking difference between the lipids from these 2 sources was that phosphatidylglycerol of lamellar organelles contained predominantly palmitic acid, whereas the phosphatidylglycerol obtained by lavage of humans contained large amounts of stearic and oleic acids.     

Taeniasis in 1994

OBJECTIVE: To present our method of sacro-spinous ligament fixation by palpation and compare it to the classical approach described by Richter. MATERIAL: In addition to the standard instruments needed for vaginal surgery, we use a Rasemond dissector, a small O'Shaugnessy dissector with smooth branches. We also use a Bengolea forceps and a monothread-nylon (Ethilon), decimal 4 suture loaded on a needle with a 30-mm curve. PATIENTS: Twenty patients underwent this procedure from 03/15/1978 to 05/19/1995. Their ages ranged from 46 to 86 years with a mean age of 64.7 years. METHOD: This was a retrospective study of the indications, results and complications associated with this technique. RESULTS: With an average follow-up of 7 years, we observed 90% success, 10% recurrences, and no complication directly attributable to this technique. This technique is valuable because of its effectiveness and simplicity. CONCLUSIONS: Sacrospinous fixation by palpation is more simple and provides the same results as the classic exposure technique. We describe the technique in this text. The efficacy of sacro-spinous ligament fixation by palpation would be improved by its systematic and bilateral use. Its value must be confirmed by a controlled prospective study to confirm our impression that our technique carries fewer risks.     

Which variable of stenosis severity best describes the significance of an isolated left anterior descending coronary artery lesion? Correlation between quantitative coronary angiography, intracoronary Doppler measurements and high dose dipyridamole echocardiography

Herein is a procedure described using a semipermeable membrane (SPM) for enrichment of organic chemicals from lipid-containing samples. Dialysis with SPMs in an organic solvent can separate organochlorine contaminants such as non-, mono-, and di-o-PCBs, PCDDs, PCDFs, PCDTs, PCNs, pesticides, and PAHs from lipids. The method is nondestructive and more than 20 g of lipid can be dialyzed in a single membrane with acceptable recoveries of the internal standards, nearly independent of amount and type of lipid dialyzed. The lipid removal process shows good consistency between subsamples, and the lipid content can be reduced by 90-99%, depending on species and amount lipid. Neither triglycerides nor phospholipids were obtained in the dialysate fraction. The utility and reliability of the method is displayed by good precision for 72 PCBs and 27 organochlorine pesticides in the concentration range 0.05-50 micrograms/sample for triplicate subsamples, by the consistency in PCDD/F levels compared to a classic analytical procedure, and by the analysis of the above listed chemicals in approximately 200 biological samples of a wide variety of types. This technique can also be used as an enrichment device for contaminants when huge amounts of lipids are extracted for toxicological studies. Moreover, it is possible to use SPM to cleanup other samples from molecules with relatively high masses, e.g. sediments, soil, compost, and tar materials.     

Exposure of anionic phospholipids upon platelet activation permits binding of beta 2 glycoprotein I and through it that of IgG antiphospholipid antibodies. Studies in platelets from patients with antiphospholipid syndrome and normal subjects

Patients with antiphospholipid syndrome, whether primary or secondary to systemic lupus erythematosus, may have thrombocytopenia. Their antibodies to anionic phospholipids might bind to phospholipids on the platelet wall but anionic phospholipids are asymmetrically located in the inner leaflet. In addition, antibodies to anionic phospholipids may require beta 2 glycoprotein I (beta 2GPI) as a cofactor in order to bind to phospholipids. In turn, beta 2GPI has high affinity for anionic phospholipids. Loss of this asymmetry occurs upon platelet activation and could thus permit such antibody-beta 2GPI-platelet interaction. We studied this by flow cytometry using purified beta 2GPI-FITC labelled and similarly labelled affinity-purified polyclonal antibodies to cardiolipin or phosphatidylserine (aPL) obtained from sera of patients with primary antiphospholipid syndrome. Five percent of resting platelets were bound by aPL in the presence of beta 2GPI. Such binding increased when we activated platelets with various agonists, reaching 31% with the concurrent use of thrombin and the calcium ionophore A23187. Platelet activation resulted in the expression of GMP140 but this did not correlate with aPL binding. This probably reflects that the expression of GMP140, which depends on their secretion of alpha granules, has different agonist responses and occurs at different times than do microvesicle formation and expression of prothrombinase activity which coincide with the loss of phospholipid asymmetry on the platelet wall. When we studied the binding of purified beta 2GPI we also found that it binds preferentially to activated platelets and that it seems to be a prerequisite for the binding of aPL onto them. Our findings indicate that aPL from patients with antiphospholipid syndrome may bind to activated platelets through beta 2GPI.     

Prediction of Columnar to Equiaxed Transition during Diffusion-Controlled Dendritic Alloy Solidification

A model is presented to predict the columnar to equiaxed transition (CET) in alloy castings. The model is based on a multiphase approach and accounts for heat and solute diffusion, as well as for grain nucleation, growth, and morphology. The model equations are applicable to both columnar and equiaxed dendritic solidification, thus offering an efficient single-domain formulation. A fixed grid, fully implicit finite-difference procedure is employed in the numerical solution, and a novel front tracking technique is incorporated that is also implicit in nature and readily applies to multidimensional situations. Calculations are performed for one-dimensional (1-D) and two-dimensional (2-D) castings of Al-Cu and Sn-Pb alloys. The calculated CET positions are compared with previous measurements in a (1-D) ingot cast under well-controlled conditions, and good agreement is found. The effects of various casting parameters on the CET are numerically explored.     

8-OH-DPAT influences extracellular levels of serotonin and dopamine in the medial preoptic area of male rats

The effect of phospholipase C treatment on cardiolipin biosynthesis was investigated in intact H9c2 cardiac myoblasts. Treatment of cells with phosphatidylcholine-specific Clostridium welchii phospholipase C reduced the pool size of phosphatidylcholine compared with controls whereas the pool size of cardiolipin and phosphatidylglycerol were unaffected. Pulse labeling experiments with [1,3-3H]glycerol and pulse-chase labeling experiments with [1,3-3H]glycerol were performed in cells incubated or pre-incubated in the absence or presence of phospholipase C. In all experiments, radioactivity incorporated into cardiolipin and phosphatidylglycerol were reduced in phospholipase C-treated cells with time compared with controls indicating attenuated de novo biosynthesis of these phospholipids. Addition of 1,2-dioctanoyl-sn-glycerol, a cell permeable 1,2-diacyl-sn-glycerol analog, to cells mimicked the inhibitory effect of phospholipase C on cardiolipin and phosphatidylglycerol biosynthesis from [1,3-3H]glycerol indicating the involvement of 1,2-diacyl-sn glycerol. The mechanism for the reduction in cardiolipin and phosphatidylglycerol biosynthesis in phospholipase C-treated cells appeared to be a decrease in the activities of phosphatidic acid:cytidine-5'triphosphate cytidylyltransferase and phosphatidylglycerolphosphate synthase, mediated by elevated 1,2-diacylsn-glycerol levels. Upon removal of phospholipase C from the incubation medium, phosphatidylcholine biosynthesis from [methyl-3H]choline was markedly stimulated. These data suggest that de novo phosphatidylglycerol and cardiolipin biosynthesis may be regulated by 1,2-diacyl-sn-glycerol and support the notion that phosphatidylglycerol and cardiolipin biosynthesis may be coordinated with phosphatidylcholine biosynthesis in H9c2 cardiac myoblast cells.     

Multicomponent diffusion simulation based on finite elements

A numerical model is presented to treat multicomponent, multiphase diffusion problems. Unlike other recent approaches that are based on the finite-difference method, analytical solutions, or particular thermodynamic models, a general procedure based on the finite-element technique is applied. The suggested formalism is based on the solution of the integral statement of the generalized diffusion equation. This treatment allows for a simple implementation of particular boundary conditions and can easily be extended from a one- to a multidimensional analysis. A brief overview of the formal representation of multicomponent diffusion coefficients, as suggested by Andersson and Agren, is given. The finite-element diffusivity matrices are evaluated for a one-dimensional bar and a two-dimensional triangular element. The model is applied to some classical examples in diffusion simulation in both one and two dimensions. The results are compared to available analytical solutions or experimental data.     

高建筑用Q345GJC-Z35特厚钢板的研制

 利用250mm连铸坯料,在3800mm宽厚板轧机上针对Q345GJC-Z35钢种进行了厚50～80mm钢板的TMCP工艺试验,确定了相应的热轧及控冷工艺条件。结果表明：采用碳的质量分数低于0.11%添加微量复合铌、钒、钛元素,按照2阶段控制,当轧到成品钢板厚度的2～3倍时开始待温,精轧开轧温度小于860℃,终轧温度为820～860℃,生产的Q345GJC-Z35高强度厚板的性能完全超出国家标准GB19879—2005要求,而且其钢板的平均断面收缩率都大于50%,远高于Z35钢板的技术要求。实现了钢板很好的强韧性匹配,工艺上不用后续热处理,减少了工艺流程,节约了成本。     

Groundwater Flow and Contaminant Transport Simulation with Imprecise Parameters

A methodology has been developed in this study wherein a genetic algorithm (GA) is used to find a global optimal solution to a groundwater flow and contaminant problem by incorporating an artificial neural network (ANN) to evaluate the objective function within the genetic algorithm. The study shows that an ANN-GA technique can be used to find the uncertainties in output parameters due to imprecision in input parameters. The ANN-GA methodology is applied to five case studies involving radial flow in a well, one-dimensional solute transport in steady uniform flow, a two-dimensional heterogeneous steady flow, a two-dimensional solute transport, and a two-dimensional unsteady groundwater flow to demonstrate the efficiency and effectiveness of the developed algorithm. The results show that, with this approach, one can successfully measure the uncertainty in groundwater flow and contaminant transport simulations and achieve a considerable reduction in computational effort when compared to the vertex method that has been widely used in the past.