Folate: Methods of analysis

Folates and its derivatives occur as polyglutamates in nature. The multiplicity of forms and the generally low levels in foods makes quantitative analysis of folate a difficult task. The assay of folates from foods generally involves three steps: liberation of folates from the cellular matrix; deconjugation from the polyglutamate to the mono and di-glutamate forms; and the detection of the biological activity or chemical concentration of the resulting folates. The detection methods used are the microbiological assay relying on the turbidimetric bacterial growth of Lactobacillus rhamnosus which is by far the most commonly used method; the HPLC and LC/MS techniques and bio-specific procedures. This review attempts to describe the methods along with the merits and demerits of using each of these methods.

Matrix effects: The Achilles heel of quantitative high-performance liquid chromatography-electrospray-tandem mass spectrometry

High-performance liquid chromatography coupled by an electrospray ion source to a tandem mass spectrometer (HPLC-EST-MS/ MS) is the current analytical method of choice for quantitation of analytes in biological matrices. With HPLC-ESI-MS/MS having the characteristics of high selectivity, sensitivity, and throughput, this technology is being increasingly used in the clinical laboratory. An important issue to be addressed in method development, validation, and routine use of HPLC-ESI-MS/MS is matrix effects. Matrix effects are the alteration of ionization efficiency by the presence of coeluting substances. These effects are unseen in the chromatograrn but have deleterious impact on methods accuracy and sensitivity. The two common ways to assess matrix effects are either by the postextraction addition method or the postcolumn infusion method. To remove or minimize matrix effects, modification to the sample extraction methodology and improved chromatographic separation must be performed. These two parameters are linked together and form the basis of developing a successful and robust quantitative HPLC-EST-MS/MS method. Due to the heterogenous nature of the population being studied, the variability of a method must be assessed in samples taken from a variety of subjects. In this paper, the major aspects of matrix effects are discussed with an approach to address matrix effects during method validation proposed. (c) 2004 The Canadian Society of Clinical Chemists. All rights reserved.

Determining the effective dimensionality of the genetic variance-covariance matrix

Determining the dimensionality of G provides an important perspective on the genetic basis of a multivariate suite of traits. Since the introduction of Fisher's geometric model, the number of genetically independent traits underlying a set of functionally related phenotypic traits has been recognized as an important factor influencing the response to selection. Here, we show how the effective dimensionality of G can be established, using a method for the determination of the dimensionality of the effect space from a multivariate general linear model introduced by AMEMIYA (1985). We compare this approach with two other available methods, factor-analytic modeling and bootstrapping, using a half-sib experiment that estimated G for eight cuticular hydrocarbons of Drosophila serrata. In our example, eight pheromone traits were shown to be adequately represented by only two underlying genetic dimensions by Amemiya's approach and factor-analytic modeling of the covariance structure at the sire level. In, contrast, bootstrapping identified four dimensions with significant genetic variance. A simulation study indicated that while the performance of Amemiya's method was more sensitive to power constraints, it performed as well or better than factor-analytic modeling in correctly identifying the original genetic dimensions at moderate to high levels of heritability. The bootstrap approach consistently overestimated the number of dimensions in all cases and performed less well than Amemiya's method at subspace recovery.

Arthritic disease suppression and cartilage protection with glycosaminoglycan polypeptide complexes (Peptacans) derived from the cartilage extracellular matrix: A novel approach to therapy

Molecular fragments of cartilage are antigenic and can stimulate an autoimmune response. Oral administration of type II collagen prevents disease onset in animal models of arthritis but the effects of other matrix components have not been reported. We evaluated glycosaminoglycan polypeptides (GAG-P) and matrix proteins (CaP) from cartilage for a) mitigating disease activity in rats with collagen-induced arthritis (CIA) and adjuvant-induced arthritis (AIA) and b) stimulating proteoglycan (PG) synthesis by chondrocytes in-vitro. CIA and AIA were established in Wistar rats using standard methods. Agents were administered orally (10–200 mg/kg), either for seven days prior to disease induction (toleragenic protocol), or continuously for 15 days after injecting the arthritigen (prophylactic protocol). Joint swelling and arthritis scores were determined on day 15. Histological sections of joint tissues were assessed post-necropsy. In chondrocyte cultures, CaP + / − interleukin-1 stimulated PG biosynthesis. CaP was also active in preventing arthritis onset at 3.3, 10 or 20 mg/kg in the rat CIA model using the toleragenic protocol. It was only active at 20 and 200 mg/kg in the CIA prophylactic protocol. GAG-P was active in the CIA toleragenic protocol at 20 mg/kg but chondroitin sulfate and glucosamine hydrochloride or glucosamine sulfate were all inactive. The efficacy of CaP in the rat AIA model was less than in the CIA model. These findings lead us to suggest that oral CaP could be used as a disease-modifying anti-arthritic drug.

A solvent-free matrix application method for matrix-assisted laser desorption/ionization imaging of small molecules

Matrix application continues to be a critical step in sample preparation for matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI). Imaging of small molecules such as drugs and metabolites is particularly problematic because the commonly used washing steps to remove salts are usually omitted as they may also remove the analyte, and analyte spreading is more likely with conventional wet matrix application methods. We have developed a method which uses the application of matrix as a dry, finely divided powder, here referred to as dry matrix application, for the imaging of drug compounds. This appears to offer a complementary method to wet matrix application for the MALDI-MSI of small molecules, with the alternative matrix application techniques producing different ion profiles, and allows the visualization of compounds not observed using wet matrix application methods. We demonstrate its value in imaging clozapine from rat kidney and 4-bromophenyl-1,4-diazabicyclo(3.2.2)nonane-4-carboxylic acid from rat brain. In addition, exposure of the dry matrix coated sample to a saturated moist atmosphere appears to enhance the visualization of a different set of molecules.

Complementary methods to investigate the development of clogging within a horizontal sub-surface flow tertiary treatment wetland

A combination of experimental methods was applied at a clogged, horizontal subsurface flow (HSSF) municipal wastewater tertiary treatment wetland (TW) in the UK, to quantify the extent of surface and subsurface clogging which had resulted in undesirable surface flow. The three dimensional hydraulic conductivity profile was determined, using a purpose made device which recreates the constant head permeameter test in-situ. The hydrodynamic pathways were investigated by performing dye tracing tests with Rhodamine WT and a novel multi-channel, data-logging, flow through Fluorimeter which allows synchronous measurements to be taken from a matrix of sampling points. Hydraulic conductivity varied in all planes, with the lowest measurement of 0.1 md1 corresponding to the surface layer at the inlet, and the maximum measurement of 1550 md1 located at a 0.4m depth at the outlet. According to dye tracing results, the region where the overland flow ceased received five times the average flow, which then vertically short-circuited below the rhizosphere. The tracer break-through curve obtained from the outlet showed that this preferential flow-path accounted for approximately 80% of the flow overall and arrived 8 h before a distinctly separate secondary flow-path. The overall volumetric efficiencyof the clogged system was 71% and the hydrology was simulated using a dual-path, dead-zone storage model. It is concluded that uneven inlet distribution, continuous surface loading and high rhizosphere resistance is responsible for the clog formation observed in this system. The average inlet hydraulic conductivity was 2 md1, suggesting that current European design guidelines, which predict that the system will reach an equilibrium hydraulic conductivity of 86 md1, do not adequately describe the hydrology of mature systems.

Modulation of interleukin-6 and matrix metalloproteinase 2 expression in human fibroblast-like synoviocytes by functional ionotropic glutamate receptors

Objective. Patients with rheumatoid arthritis (RA) have increased concentrations of the amino acid glutamate in synovial fluid. This study was undertaken to determine whether glutamate receptors are expressed in the synovial joint, and to determine whether activation of glutamate receptors on human synoviocytes contributes to RA disease pathology. Methods. Glutamate receptor expression was examined in tissue samples from rat knee joints and in human fibroblast-like synoviocytes (FLS). FLS from 5 RA patients and 1 normal control were used to determine whether a range of glutamate receptor antagonists influenced expression of the proinflammatory cytokine interleukin-6 (IL-6), enzymes involved in matrix degradation and cytokine processing (matrix metalloproteinase 2 [MMP-2] and MMP-9), and the inhibitors of these enzymes (tissue inhibitor of metalloproteinases 1 [TIMP-1] and TIMP-2). IL-6 concentrations were determined by enzyme-linked immunosorbent assay, MMP activity was measured by gelatin zymography, and TIMP activity was determined by reverse zymography. Fluorescence imaging of intracellular calcium concentrations in live RA FLS stimulated with specific antagonists was used to reveal functional activation of glutamate receptors that modulated IL-6 or MMP-2. Results. Ionotropic and metabotropic glutamate receptor subunit mRNA were expressed in the patella, fat pad, and meniscus of the rat knee and in human articular cartilage. Inhibition of N-methyl-D-aspartate (NMDA) receptors in RA FLS increased proMMP-2 release, whereas non-NMDA ionotropic glutamate receptor antagonists reduced IL-6 production by these cells. Stimulation with glutamate, NMDA, or kainate (KA) increased intracellular calcium concentrations in RA FLS, demonstrating functional activation of specific ionotropic glutamate receptors. Conclusion. Our findings indicate that activation of NMDA and KA glutamate receptors on human synoviocytes may contribute to joint destruction by increasing IL-6 expression. © 2007, American College of Rheumatology.

Investigating the Relations of Attacks, Methods and Objects in Regard to Information Security in Network TCP/IP Environment

Possibilities for investigations of 43 varieties of file formats (objects), joined in 10 groups; 89 information attacks, joined in 33 groups and 73 methods of compression, joined in 10 groups are described in the paper. Experimental, expert, possible and real relations between attacks’ groups, method’ groups and objects’ groups are determined by means of matrix transformations and the respective maximum and potential sets are defined. At the end assessments and conclusions for future investigation are proposed.

An Iterative Method for the Matrix Principal n-th Root

In this paper we give an iterative method to compute the principal n-th root and the principal inverse n-th root of a given matrix. As we shall show this method is locally convergent. This method is analyzed and its numerical stability is investigated.

Distance Matrix Approach to Content Image Retrieval

As the volume of image data and the need of using it in various applications is growing significantly in the last days it brings a necessity of retrieval efficiency and effectiveness. Unfortunately, existing indexing methods are not applicable to a wide range of problem-oriented fields due to their operating time limitations and strong dependency on the traditional descriptors extracted from the image. To meet higher requirements, a novel distance-based indexing method for region-based image retrieval has been proposed and investigated. The method creates premises for considering embedded partitions of images to carry out the search with different refinement or roughening level and so to seek the image meaningful content.

Quasi-Monte Carlo Methods for some Linear Algebra Problems. Convergence and Complexity

We present quasi-Monte Carlo analogs of Monte Carlo methods for some linear algebra problems: solving systems of linear equations, computing extreme eigenvalues, and matrix inversion. Reformulating the problems as solving integral equations with a special kernels and domains permits us to analyze the quasi-Monte Carlo methods with bounds from numerical integration. Standard Monte Carlo methods for integration provide a convergence rate of O(N^(−1/2)) using N samples. Quasi-Monte Carlo methods use quasirandom sequences with the resulting convergence rate for numerical integration as good as O((logN)^k)N^(−1)). We have shown theoretically and through numerical tests that the use of quasirandom sequences improves both the magnitude of the error and the convergence rate of the considered Monte Carlo methods. We also analyze the complexity of considered quasi-Monte Carlo algorithms and compare them to the complexity of the analogous Monte Carlo and deterministic algorithms.

Algorithmic Methods in Queues and in the Exploration of Point Processes

This is a review of methodology for the algorithmic study of some useful models in point process and queueing theory, as discussed in three lectures at the Summer Institute at Sozopol, Bulgaria. We provide references to sources where the extensive details of this work are found. For future investigation, some open problems and new methodological approaches are proposed.

Three New Methods for Computing Subresultant Polynomial Remainder Sequences (PRS’S)

Given the polynomials f, g ∈ Z[x] of degrees n, m, respectively, with n > m, three new, and easy to understand methods — along with the more efficient variants of the last two of them — are presented for the computation of their subresultant polynomial remainder sequence (prs). All three methods evaluate a single determinant (subresultant) of an appropriate sub-matrix of sylvester1, Sylvester’s widely known and used matrix of 1840 of dimension (m + n) × (m + n), in order to compute the correct sign of each polynomial in the sequence and — except for the second method — to force its coefficients to become subresultants. Of interest is the fact that only the first method uses pseudo remainders. The second method uses regular remainders and performs operations in Q[x], whereas the third one triangularizes sylvester2, Sylvester’s little known and hardly ever used matrix of 1853 of dimension 2n × 2n. All methods mentioned in this paper (along with their supporting functions) have been implemented in Sympy and can be downloaded from the link http://inf-server.inf.uth.gr/~akritas/publications/subresultants.py

A general Approach to Methods with a Sparse Jacobian for Solving Nonlinear Systems of Equations

2000 Mathematics Subject Classification: 65H10.

Fogalmak, módszerek. Páros összehasonlításokon alapuló rangsorolási módszerek (Ranking methods based on paired comparisons)

A páronként összehasonlított alternatívák rangsorolásának problémája egyaránt felmerül a szavazáselmélet, a statisztika, a tudománymetria, a pszichológia és a sport területén. A nemzetközi szakirodalom alapján részletesen áttekintjük a megoldási lehetőségeket, bemutatjuk a gyakorlati alkalmazások során fellépő kérdések kezelésének, a valós adatoknak megfelelő matematikai környezet felépítésének módjait. Kiemelten tárgyaljuk a páros összehasonlítási mátrix megadását, az egyes pontozási eljárásokat és azok kapcsolatát. A tanulmány elméleti szempontból vizsgálja a Perron-Frobenius tételen alapuló invariáns, fair bets, PageRank, valamint az irányított gráfok csúcsainak rangsorolásra javasolt internal slackening és pozíciós erő módszereket. A közülük történő választáshoz az axiomatikus megközelítést ajánljuk, ennek keretében bemutatjuk az invariáns és a fair bets eljárások karakterizációját, és kitérünk a módszerek vitatható tulajdonságaira. _____ The ranking of the alternatives or selecting the best one are fundamental issues of social choice theory, statistics, psychology and sport. Different solution concepts, and various mathematical models of applications are reviewed based on the international literature. We are focusing on the de¯nition of paired comparison matrix, on main scoring procedures and their relation. The paper gives a theoretical analysis of the invariant, fair bets and PageRank methods, which are founded on Perron-Frobenius theorem, as well as the internal slackening and positional power procedures used for ranking the nodes of a directed graph. An axiomatic approach is proposed for the choice of an appropriate method. Besides some known characterizations for the invariant and fair bets methods, we also discuss the violation of some properties, meaning their main weakness.