Characterization of allergoids from ovalbumin in vitro and in vivo

Several in vivo and in vitro methods for monitoring immunological properties of two allergoids obtained by formaldehyde treatment of ovalbumin (OA) were developed. The calculated molecular weight of allergoids was 80 kD (OA-F1) and 165 kD (OA-F2), respectively. The allergenic activity in vitro of allergoids in mast-cell histamine release assay was 1000 times lower than of OA. Both allergoids showed reduced ability to induce passive cutaneous anaphylaxis in the Sprague-Dawley rats or systemic anaphylaxis in Dunkin-Harley guinea-pigs. The ability of OA and allergoids to bind to the OA-specific IgE antibodies was measured in vivo by the inhibition of passive cutaneous anaphylaxis (PCA-inhibition). Allergoid binding to IgE was 51-66% lower than the native allergen. Moreover, the avidity of OA-specific IgG antibodies, measured by ELISA-inhibition, for allergoids and allergen was of the same order. Allergoids induced a different pattern of humoral immune response from that, induced by the native allergen. Thus, after immunization of BALB/c mouse, both allergoids induced a higher production of IgG and a lower production of IgE than OA, only OA-F2 induced a lower production of IgG1. The differences in the IgA response to the immunogens was not significant. Delayed hypersensitivity studies in the BALB/c mouse showed that allergoids were 5- to 12-times less effective in inducing a cell-mediated immune response than OA. The present study provides a battery of immunological methods for preclinical testing of modified allergens.     

Hull-less Barley Varieties: Storage Proteins and Amino Acid Distribution in Relation to Nutritional Quality

Barley (Hordeum vulgare L.) has had an important impact on human nutrition. Hull-less barley is a genetically improved type that has been widely used in recent years. Six Brazilian hull-less barley varieties (IAC-IBON 214-82; IAC 8612-421; IAC 8501-31; IAC 8501-12; IAPAR 39-ACUMAI; IAC 8501-22) were analyzed for storage protein constituents, amino acid contents, and similarity among the hull-less barley varieties. Albumins, globulins, prolamins I and II, and glutelins were extracted and separated by SDS-PAGE. The total protein amino acid contents of the flour were also determined for each variety by TLC and HPLC. Variations in intensity and appearance and disappearance of protein bands were observed among the varieties suggesting genetic variability. However, the amino acid profile did not indicate any major variations in the amino acid concentrations. The high lysine and threonine total concentrations detected in the seeds of the hull-less barley varieties encouraged an investigation into the regulation of amino acid metabolism and storage protein synthesis.     

Screening of supports for the immobilization of pectinmethylesterase from acerola (Malpighia glabra L)

A partially purified extract of pectinmethylesterase (PME) from acerola fruit was immobilized on various supports: glass, celite, chrysotile, agarose, concanavalin A Sepharose 4B, egg shell, polyacrylamide and gelatin. In addition, reticulation with glutaraldehyde was assessed, as well as the use of gelatin in the presence of celite, glass and silica. The highest immobilization yields were obtained when the pectinmethylesterase was immobilized in concanavalin A Sepharose 4B (81.7%) and in gelatin‐water (78.0%). Copyright © 2004 Society of Chemical Industry     

Resistance to vasopressin action on the kidney in patients with Cushing's disease

OBJECTIVE: To assess the plasma levels and action of arginine vasopressin (AVP) in patients with Cushing's disease. There are many reports that patients with Addison's disease have increased AVP levels associated with hyponatraemia and hypoosmolality, but none on the dynamics of secretion of this neurohormone during osmolality-based stimulation in patients with chronic hypercortisolism. DESIGN AND SUBJECTS: The plasma AVP concentration and the urinary and plasma osmolality after a 7.5-h water deprivation test (WDT) were evaluated in 13 patients with Cushing's disease and 15 normal (control) individuals. In patients with Cushing's disease we also assessed the urinary osmolality in response to 10 micrograms i.v. desmopressin (DDAVP) administered at the end of the WDT. RESULTS: At the end of the WDT, urinary osmolality was significantly lower in patients with Cushing's disease (511.5 +/- 148.5 mOsm/l) than in the normal subjects (981.1 +/- 107.1 mOsm/l, P < 0.001), whereas plasma osmolality did not differ between the two groups. Consequently, the urine/plasma osmolality ratio (Uosm/Posm) was lower in patients with Cushing's disease than in normal individuals (1.8 +/- 0.5 compared with 3.4 +/- 0.4, P < 0.001). The AVP concentration also was greater (7.3 +/- 3.1 pmol/l) in those with Cushing's disease than in the controls (3.9 +/- 2.3 pmol/l, P < 0.005). After administration of DDAVP to the hypercortisolaemic patients, the urinary osmolality attained (718.0 +/- 200.0 mOsm/l) was still lower than that in the normal group at the end of WDT (P < 0.005). CONCLUSIONS: Patients with Cushing's disease presented higher AVP levels and smaller Uosm/Posm ratios than normal subjects. After DDAVP, the patients with Cushing's disease were unable to concentrate the urine adequately. These data suggest that the kidney shows resistance to the action of both endogenous and exogenous AVP in patients with Cushing's disease.     

Cutting force estimation in sculptured surface milling

Cutting force milling models developed up to now are mostly used for planar milling using end-mills. Only a reduced number of models applying ball-end mills have been developed. Furthermore these models usually only consider horizontal surface machining, even though the main application of ball-end mills is sculptured surface machining. This article proposes a model that is able to estimate the cutting forces in inclined surfaces machined both up-milling and down-milling. For this purpose a semi-mechanistic model has been developed that calculates the cutting forces based on a set of coefficients which depend on the material, the tool, the cutting conditions, the machining direction and the slope of the surface.A coordinate transformation has been included in order to consider the slope milling case with different cutting directions.The model has been tested on two materials, an aluminum alloy Al7075-T6 and a 52 HRC tool steel AISI H13. Validation tests have been carried out on inclined planes using different slopes and different machining directions.The results provide errors below 10% in most of the cases and both the value and shape of the predicted forces adjust the measured cutting force.     

Real-time lane tracking using Rao-Blackwellized particle filter

A novel approach to real-time lane modeling using a single camera is proposed. The proposed method is based on an efficient design and implementation of a particle filter which applies the concepts of the Rao-Blackwellized particle filter (RBPF) by separating the state into linear and non-linear parts. As a result the dimensionality of the problem is reduced, which allows the system to perform in real-time in embedded systems. The method is used to determine the position of the vehicle inside its own lane and the curvature of the road ahead to enhance the performance of advanced driver assistance systems. The effectiveness of the method has been demonstrated implementing a prototype and testing its performance empirically on road sequences with different illumination conditions (day and nightime), pavement types, traffic density, etc. Results show that our proposal is capable of accurately determining if the vehicle is approaching the lane markings (Lane Departure Warning), and the curvature of the road ahead, achieving processing times below 2 ms per frame for laptop CPUs, and 12 ms for embedded CPUs.     

Performance bounds in linear control of unstable MIMO systems with pole location constraint

This paper proposes a methodology to compute quadratic performance bounds when the closed loop poles of a discrete-time multivariable control loop are confined to a disk, centred at the origin, and with radius less than one. The underlying philosophy in this constraint is to avoid certain undesirable dynamic features which arise in quadratic optimal designs. An expression for the performance loss due to the pole location constraint is also provided. Using numerical examples, we show that the performance loss is compensated by an improved transient performance, specially visible in the control signals.     

Design Optimization of Composite Prosthetic Tubes Using GA-ANN Algorithm Considering Tsai-Wu Failure Criteria

The investigation of possible failures in composite materials is a matter of very great importance, and the Tsai-Wu criterion is an effective criterion for analyzing those flaws in anisotropic materials and defining whether the material at a given load will or will not suffer structural failure. In this study, an optimization procedure is proposed to minimize the maximum value of Tsai-Wu of laminated composite tubes subject to axial loading. Artificial neural networks and genetic algorithms are chosen as optimization tools. The results of this study show that the developed algorithm converges faster. Then, the maximum Tsai-Wu value is used as the objective function and the fiber orientations are the constraints in the optimization process. The results yielded by them are compared and discussed. Optimal results are compared with respect to the usual initial design. The design approach is recommended for structures where composites are the key load-carrying members such as orthopedic prosthesis.     

MIL-100(Fe) Sub-Micrometric Capsules as a Dual Drug Delivery System

Nanoparticles of metal–organic frameworks (MOF NPs) are crystalline hybrid micro- or mesoporous nanomaterials that show great promise in biomedicine due to their significant drug loading ability and controlled release. Herein, we develop porous capsules from aggregate of nanoparticles of the iron carboxylate MIL-100(Fe) through a low-temperature spray-drying route. This enables the concomitant one-pot encapsulation of high loading of an antitumor drug, methotrexate, within the pores of the MOF NPs, and the collagenase enzyme (COL), inside the inter-particular mesoporous cavities, upon the formation of the capsule, enhancing tumor treatment. This association provides better control of the release of the active moieties, MTX and collagenase, in simulated body fluid conditions in comparison with the bare MOF NPs. In addition, the loaded MIL-100 capsules present, against the A-375 cancer cell line, selective toxicity nine times higher than for the normal HaCaT cells, suggesting that MTX@COL@MIL-100 capsules may have potential application in the selective treatment of cancer cells. We highlight that an appropriate level of collagenase activity remained after encapsulation using the spray dryer equipment. Therefore, this work describes a novel application of MOF-based capsules as a dual drug delivery system for cancer treatment.