Design and Synthesis of New Quinazolin-4-one Derivatives with Negative mGlu7 Receptor Modulation Activity and Antipsychotic-Like Properties.

Following the glutamatergic theory of schizophrenia and based on our previous study regarding the antipsychotic-like activity of mGlu7 NAMs, we synthesized a new compound library containing 103 members, which were examined for NAM mGlu7 activity in the T-REx 293 cell line expressing a recombinant human mGlu7 receptor. Out of the twenty-two scaffolds examined, active compounds were found only within the quinazolinone chemotype. 2-(2-Chlorophenyl)-6-(2,3-dimethoxyphenyl)-3-methylquinazolin-4(3H)-one (A9-7, ALX-171, mGlu7 IC50 = 6.14 µM) was selective over other group III mGlu receptors (mGlu4 and mGlu8), exhibited satisfactory drug-like properties in preliminary DMPK profiling, and was further tested in animal models of antipsychotic-like activity, assessing the positive, negative, and cognitive symptoms. ALX-171 reversed DOI-induced head twitches and MK-801-induced disruptions of social interactions or cognition in the novel object recognition test and spatial delayed alternation test. On the other hand, the efficacy of the compound was not observed in the MK-801-induced hyperactivity test or prepulse inhibition. In summary, the observed antipsychotic activity profile of ALX-171 justifies the further development of the group of quinazolin-4-one derivatives in the search for a new drug candidate for schizophrenia treatment.

New 1,2,4-oxadiazole derivatives with positive mGlu4 receptor modulation activity and antipsychotic-like properties.

Considering the allosteric regulation of mGlu receptors for potential therapeutic applications, we developed a group of 1,2,4-oxadiazole derivatives that displayed mGlu4 receptor positive allosteric modulatory activity (EC50 = 282-656 nM). Selectivity screening revealed that they were devoid of activity at mGlu1, mGlu2 and mGlu5 receptors, but modulated mGlu7 and mGlu8 receptors, thus were classified as group III-preferring mGlu receptor agents. None of the compounds was active towards hERG channels or in the mini-AMES test. The most potent in vitro mGlu4 PAM derivative 52 (N-(3-chloro-4-(5-(2-chlorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)picolinamide) was readily absorbed after i.p. administration (male Albino Swiss mice) and reached a maximum brain concentration of 949.76 ng/mL. Five modulators (34, 37, 52, 60 and 62) demonstrated significant anxiolytic- and antipsychotic-like properties in the SIH and DOI-induced head twitch test, respectively. Promising data were obtained, especially for N-(4-(5-(2-chlorophenyl)-1,2,4-oxadiazol-3-yl)-3-methylphenyl)picolinamide (62), whose effects in the DOI-induced head twitch test were comparable to those of clozapine and better than those reported for the selective mGlu4 PAM ADX88178.

Sampling Points-Independent Identification of the Fractional Maxwell Model of Viscoelastic Materials Based on Stress Relaxation Experiment Data.

Considerable development has been observed in the area of applying fractional-order rheological models to describe the viscoelastic properties of miscellaneous materials in the last few decades together with the increasingly stronger adoption of fractional calculus. The fractional Maxwell model is the best-known non-integer-order rheological model. A weighted least-square approximation problem of the relaxation modulus by the fractional Maxwell model is considered when only the time measurements of the relaxation modulus corrupted by additive noises are accessible for identification. This study was dedicated to the determination of the model, optimal in the sense of the integral square weighted model quality index, which does not depend on the particular sampling points applied in the stress relaxation experiment. It is proved that even when the real description of the material relaxation modulus is entirely unknown, the optimal fractional Maxwell model parameters can be recovered from the relaxation modulus measurements recorded for sampling time points selected randomly according to respective randomization. The identified model is a strongly consistent estimate of the desired optimal model. The exponential convergence rate is demonstrated both by the stochastic convergence analysis and by the numerical studies. A simple scheme for the optimal model identification is given. Numerical studies are presented for the materials described by the short relaxation times of the unimodal Gauss-like relaxation spectrum and the long relaxation times of the Baumgaertel, Schausberger and Winter spectrum. These studies have shown that the appropriate randomization introduced in the selection of sampling points guarantees that the sequence of the optimal fractional Maxwell model parameters asymptotically converge to parameters independent of these sampling points. The robustness of the identified model to the measurement disturbances was demonstrated by analytical analysis and numerical studies.

A Class of Algorithms for Recovery of Continuous Relaxation Spectrum from Stress Relaxation Test Data Using Orthonormal Functions.

The viscoelastic relaxation spectrum provides deep insights into the complex behavior of polymers. The spectrum is not directly measurable and must be recovered from oscillatory shear or relaxation stress data. The paper deals with the problem of recovery of the relaxation spectrum of linear viscoelastic materials from discrete-time noise-corrupted measurements of relaxation modulus obtained in the stress relaxation test. A class of robust algorithms of approximation of the continuous spectrum of relaxation frequencies by finite series of orthonormal functions is proposed. A quadratic identification index, which refers to the measured relaxation modulus, is adopted. Since the problem of relaxation spectrum identification is an ill-posed inverse problem, Tikhonov regularization combined with generalized cross-validation is used to guarantee the stability of the scheme. It is proved that the accuracy of the spectrum approximation depends both on measurement noises and the regularization parameter and on the proper selection of the basis functions. The series expansions using the Laguerre, Legendre, Hermite and Chebyshev functions were studied in this paper as examples. The numerical realization of the scheme by the singular value decomposition technique is discussed and the resulting computer algorithm is outlined. Numerical calculations on model data and relaxation spectrum of polydisperse polymer are presented. Analytical analysis and numerical studies proved that by choosing an appropriate model through selection of orthonormal basis functions from the proposed class of models and using a developed algorithm of least-square regularized identification, it is possible to determine the relaxation spectrum model for a wide class of viscoelastic materials. The model is smoothed and robust on measurement noises; small model approximation errors are obtained. The identification scheme can be easily implemented in available computing environments.

On Applicability of the Relaxation Spectrum of Fractional Maxwell Model to Description of Unimodal Relaxation Spectra of Polymers.

The relaxation time and frequency spectra are vital for constitutive models and for insight into the viscoelastic properties of polymers, since, from the spectra, other material functions used to describe rheological properties of various polymers can be uniquely determined. In recent decades the non-integer order differential equations have attracted interest in the description of time-dependent processes concerning relaxation phenomena. The fractional Maxwell model (FMM) is probably the most known rheological model of non-integer order. However, the FMM spectrum has not yet been studied and used to describe rheological materials. Therefore, the goal of the present paper was to study the applicability of the relaxation spectrum of FMM to the description of the relaxation spectra of polymers. Based on the known integral representation of the Mittag-Leffler two-parameter function, analytical formulas describing relaxation time and frequency spectra of FMM model were derived. Monotonicity of the spectra was analyzed and asymptotic properties were established. Relaxation frequency spectrum grows for large frequencies with a positive power law, while the relaxation time spectrum decays for large times with a negative power of time. Necessary and sufficient conditions for the existence of the local extrema of the relaxation spectra were derived in the form of two trigonometric inequalities. A simple procedure for checking the existence or absence of the spectra extrema was developed. Direct analytical formulas for the local extrema, minima, and maxima are given in terms of model fractional and viscoelastic parameters. The fractional model parameters, non-integer orders of the stress and strain derivatives of FMM uniquely determine the existence of the spectrum extrema. However, the viscoelastic parameters of the FMM, elastic modulus, and relaxation time affect the maxima and minima of the relaxation spectra and the values of their local peaks. The influence of model parameters on their local extrema was examined. Next, the applicability of the continuous-discrete spectrum of FMM to describe Baumgaertel, Schausberger and Winter (BSW) and unimodal Gauss-like relaxation spectra, commonly used to describe rheological properties of various polymers, was examined. Numerical experiments have shown that by respective choice of the FMM parameters, in particular by respective choice of the orders of fractional derivatives of the stress and strain, a good fit for the relaxation modulus experiment data was obtained for polymers characterized both by BSW and Gauss-like relaxation spectra. As a result, a good approximation of the real spectra was reached. Thus, the viscoelastic relaxation spectrum of FMM, due to the availability of the two extra degrees of freedom (non-integer orders of the stress and strain derivatives), provides deep insights into the complex behavior of polymers and can be applied for a wide class of polymers with unimodal relaxation spectra.

Two-Level Scheme for Identification of the Relaxation Time Spectrum Using Stress Relaxation Test Data with the Optimal Choice of the Time-Scale Factor.

The viscoelastic relaxation spectrum is vital for constitutive models and for insight into the mechanical properties of materials, since, from the relaxation spectrum, other material functions used to describe rheological properties can be uniquely determined. The spectrum is not directly accessible via measurement and must be recovered from relaxation stress or oscillatory shear data. This paper deals with the problem of the recovery of the relaxation time spectrum of linear viscoelastic material from discrete-time noise-corrupted measurements of a relaxation modulus obtained in the stress relaxation test. A two-level identification scheme is proposed. In the lower level, the regularized least-square identification combined with generalized cross-validation is used to find the optimal model with an arbitrary time-scale factor. Next, in the upper level, the optimal time-scale factor is determined to provide the best fit of the relaxation modulus to experiment data. The relaxation time spectrum is approximated by a finite series of power-exponential basis functions. The related model of the relaxation modulus is proved to be given by compact analytical formulas as the products of power of time and the modified Bessel functions of the second kind. The proposed approach merges the technique of an expansion of a function into a series of independent basis functions with the least-squares regularized identification and the optimal choice of the time-scale factor. Optimality conditions, approximation error, convergence, noise robustness and model smoothness are studied analytically. Applicability ranges are numerically examined. These studies have proved that using a developed model and algorithm, it is possible to determine the relaxation spectrum model for a wide class of viscoelastic materials. The model is smoothed and noise robust; small model errors are obtained for the optimal time-scale factors. The complete scheme of the hierarchical computations is outlined, which can be easily implemented in available computing environments.

How to Make the Stress Relaxation Experiment for Polymers More Informative.

Different viscoelastic models and characteristics are commonly used to describe, analyze, compare and improve the mechanical properties of polymers. A time-dependent linear relaxation modulus next to frequency-domain storage and loss moduli are the basic rheological material functions of polymers. The exponential Maxwell model and the exponential stretched Kohlrausch-Williams-Watts model are, probably, the most known linear rheological models of polymers. There are different identification methods for such models, some of which are dedicated to specific models, while others are general in nature. However, the identification result, i.e., the best model, always depends on the specific experimental data on the basis of which it was determined. When the rheological stress relaxation test is performed, the data are composed of the sampling instants used in the test and on the measurements of the relaxation modulus of the real material. To build a relaxation modulus model that does not depend on sampling instants is a fundamental concern. The problem of weighted least-squares approximation of the real relaxation modulus is discussed when only the noise-corrupted time-measurements of the relaxation modulus are accessible for identification. A wide class of models, that are continuous, differentiable and Lipschitz with respect to parameters, is considered for the relaxation modulus approximation. The main results concern the models that are selected asymptotically as the number of measurements tends to infinity. It is shown that even when the true relaxation modulus description is completely unknown, the approximate optimal model parameters can be derived from the measurement data that are obtained for sampling instants that are selected randomly due to the appropriate randomization introduced whenever certain conditions regarding the adopted class of models are satisfied. It is shown that the most commonly used stress relaxation models, the Maxwell and Kohlrausch-Williams-Watts models, satisfy these conditions. Since the practical problems of the identification of relaxation modulus models are usually ill posed, Tikhonov regularization is applied to guarantee the stability of the regularized solutions. The approximate optimal model is a strongly consistent estimate of the regularized model that is optimal in the sense of the deterministic integral weighted square error. An identification algorithm leading to the best regularized model is presented. The stochastic-type convergence analysis is conducted for noise-corrupted relaxation modulus measurements, and the exponential convergence rate is proved. Numerical studies for different models of the relaxation modulus used in the polymer rheology are presented for the material described by a bimodal Gauss-like relaxation spectrum. Numerical studies have shown that if appropriate randomization is introduced in the selection of sampling instants, then optimal regularized models of the relaxation modulus being asymptotically independent of these time instants can be recovered from the stress relaxation experiment data. The robustness of the identification algorithm to measurement noises was demonstrated both by analytical and numerical analyses.

On Recovery of a Non-Negative Relaxation Spectrum Model from the Stress Relaxation Test Data.

The relaxation spectra, from which other material functions used to describe mechanical properties of materials can be uniquely determined, are important for modeling the rheological properties of polymers used in chemistry, food technology, medicine, cosmetics, and many other industries. The spectrum, being not directly accessible by measurement, is recovered from relaxation stress or oscillatory shear data. Only a few models and identification methods take into account the non-negativity of the real spectra. In this paper, the problem of recovery of non-negative definite relaxation spectra from discrete-time noise-corrupted measurements of relaxation modulus obtained in the stress relaxation test is considered. A new hierarchical identification scheme is developed, being applicable both for relaxation time and frequency spectra. Finite-dimensional parametric classes of models are assumed for the relaxation spectra, described by a finite series of power-exponential and square-exponential basis functions. The related models of relaxation modulus are given by compact analytical formula, described by the products of power of time and the modified Bessel functions of the second kind for the time spectrum, and by recurrence formulas based on products of power of time and complementary error functions for frequency spectrum. The basis functions are non-negative. In result, the identification task was reduced to a finite-dimensional linear-quadratic problem with non-negative unknown model parameters. To stabilize the solution, an additional smoothing constraint is introduced. Dual approach was used to solve the stated optimal identification task resulting in the hierarchical two-stage identification scheme. In the first stage, dual problem is solved in two levels and the vector of non-negative model parameters is computed to provide the best fit of the relaxation modulus to experiment data. Next, in second stage, the optimal non-negative spectrum model is determined. A complete scheme of the hierarchical computations is outlined; it can be easily implemented in available computing environments. The model smoothness is analytically studied, and the applicability ranges are numerically examined. The numerical studies have proved that using developed models and algorithm, it is possible to determine non-negative definite unimodal and bimodal relaxation spectra for a wide class of polymers. However, the examples also demonstrated that if the basis functions are non-negative and the model is properly selected for a given type of the real spectrum (unimodal, multimodal), the optimal model determined without non-negativity constraint can be non-negative in the dominant range of its arguments, especially in the wide neighborhood of the spectrum peaks.

Conformational analysis of cis-3, 5-dioxa-bicyclo[5.4.0]undecane and its 4-substituted derivatives by DNMR, molecular modeling, and GIAO/DFT methods.

Temperature-dependent (1)H and (13)C-NMR spectra of the title compounds are presented. Coalescence effects are discussed and assigned to dynamic process--the interconversion of bicyclic system. The free energies of activation covered the range 39-52 kJ/mol. The dioxepane ring adopts twist-chair (TC) conformation. GIAO/DFT calculation of isotropic shieldings for the set of low-energy conformations showed that only one conformer is present at 298 K in solution that matched well with experimental data.

Patterns of host use by brood parasitic Maculinea butterflies across Europe.

The range of hosts exploited by a parasite is determined by several factors, including host availability, infectivity and exploitability. Each of these can be the target of natural selection on both host and parasite, which will determine the local outcome of interactions, and potentially lead to coevolution. However, geographical variation in host use and specificity has rarely been investigated. Maculinea (= Phengaris) butterflies are brood parasites of Myrmica ants that are patchily distributed across the Palæarctic and have been studied extensively in Europe. Here, we review the published records of ant host use by the European Maculinea species, as well as providing new host ant records for more than 100 sites across Europe. This comprehensive survey demonstrates that while all but one of the Myrmica species found on Maculinea sites have been recorded as hosts, the most common is often disproportionately highly exploited. Host sharing and host switching are both relatively common, but there is evidence of specialization at many sites, which varies among Maculinea species. We show that most Maculinea display the features expected for coevolution to occur in a geographic mosaic, which has probably allowed these rare butterflies to persist in Europe. This article is part of the theme issue 'The coevolutionary biology of brood parasitism: from mechanism to pattern'.

The Influence of L-Carnitine on Oxidative Modification of LDL In Vitro.

Owing to their structure and function, low-density lipoproteins (LDLs) are particularly susceptible to the oxidative modifications. To prevent against oxidative modification of LDL, L-carnitine, with endogenous small water-soluble quaternary amine possessing antioxidative properties, was used. The aim of this paper was to prove the in vitro influence of L-carnitine on the degree of oxidative modification of the lipid part (estimated by conjugated dienes, lipid hydroperoxides, and malondialdehyde levels) and the protein part (estimated by dityrosine and tryptophan levels) of LDL native and oxidized by cooper ions. The level of lipophylic LDL antioxidant-alpha-tocopherol was also measured.Oxidation of LDL by Cu(2+) enhanced lipid peroxidation. That was manifested by a statistically significant increase in the content of malondialdehyde (threefold), conjugated dienes (up to about 30%), and lipid hydroperoxides (up to about 50%). Cu(2+) ions were also the cause of oxidative modifications of the protein part of LDLs. It was manifested by a significant increase in dityrosine (by about 50%), whereas the level of tryptophan was significantly decreased threefold in relation to native LDL. Incubation of LDL with Cu(2+) ions also caused a significant sixfold decrease of alpha-tocopherol content in oxidized LDL. However, L-carnitine caused a decrease in the level of conjugated dienes, lipid hydroperoxide, malondialdehyde, and dityrosine by about 20% to 30%, and a significant increase (by about 50%) in the content of tryptophan in comparison with oxidative LDL and in a smaller degree significant changes with native LDL. Additionally, L-carnitine caused a significant twofold increase in alpha-tocopherol content in oxidized LDL.The above results indicate that L-carnitine protects the lipid as well as protein part of LDL particles against oxidative modifications, and this natural antioxidant might be used to prevent against diseases of oxidative origin.

Amifostine-antioxidant drug in anticancer therapy.

Antineoplastic drugs and/or their metabolites are very reactive chemically and easily react with cell components, both those neoplastic and healthy, which leads to uncontrolled cell damage. Research is being carried out to discover new drugs preventing those destructive changes in healthy cells. Amifostine is one of the few drugs with protective abilities in regard to healthy tissues during antineoplastic therapy. Amifostine alone is inactive chemically, but active metabolite amifostine WR-1065 has been assumed to protect the healthy tissues during antineoplastic therapy by bounding of anticancer drug, causing their detoxification and/or eliminating the free radicals generated during radiation and cytostatic therapy. WR-1065 also protects the healthy tissues by inhibition of apoptosis caused by irradiation and/or alkylating compounds. However, the protective mechanism of amifostine influence on healthy cells has not been fully comprehended, and the research is still being carried out.

[Clinical evaluation of periodontium in pregnant women with risk of preterm birth]. / Ocena kliniczna przyzebia kobiet w przebiegu ciazy zagrozonej.

OBJECTIVES: The aim of the study was to evaluate condition of the periodontium in pregnant women with pathological progress of the pregnancy, clinically and to compare it to periodontium in pregnant women in good health. DESIGN: Over the last years, the studies have described that periodontitis caused by dental plaque, could be the risk factor for preterm birth and low birth weight. MATERIALS AND METHODS: This study was performed in 80 pregnant women, 40 with pathologic pregnancy and 40 with normal pregnancy in it. Periodontal Indexes were used to evaluate periodontium. RESULTS: In the searching group gingivitis gravidarum haemorrhagica diffusa and hyperplastica generalisata were dominating. In the control group gingivitis gravidatum simplex and hyperplastica localisata were observed. CONCLUSIONS: More severe manifestation of gingivitis gravidarum was noticed in pregnant women with risk of preterm low birth. We did not prove correlation between amount of bacterial dental plaque in pregnant women and risk of preterm low birth weight.

[Free-living amoebae as vehicles of pathogenic bacteria]. / Pelzaki wolno zyjace jako nosiciele patogenicznych bakterii.

The free-living amoebae are ubiquitous organisms. They are found in humid soil and all water reservoirs, i.e. fresh, sea, freezing and hot water. They mainly feed on bacteria. Pathogenic properties of amoebae and the mechanisms underlying pathological changes induced during human infection have not yet been fully elucidated. They are the causative agents of primary amoebic meningo-encephalitis (PAM), granulomatous amebic encephalitis (GAE), a chronic progressive disease of the central nervous system, amebic keratitis (AK), a chronic eye infection; amebic pneumitis (AP), a chronic lung infection, and skin infection. Only a few isolates are strongly and permanently pathogenic to humans. Some isolates lose their pathogenic properties after one passage. It has been assumed that such "temporary", unstable pathogenic properties of the amoebae may be caused by internal factors carried by them. It is generally known that the free-living amoebae may be naturally infected with pathogenic bacteria, which have the ability to survive for a long time and to proliferate in the amoebae cells. The role of the amoeba in the process of maintaining, propagating and transmitting human pathogens has not been well recognized. It has been suggested that some infections can be acquired by inhaling aerosols containing amoebae cells filled with bacteria. The presence of bacteria inside the free-living amoebae possess a great challenge to organisations responsible for testing and inspecting the quality and cleanliness of surface waters, swimming pools and drinking water intakes.

Anisotropy, inhomogeneity, and tension-compression nonlinearity of human glenohumeral cartilage in finite deformation.

The tensile and compressive properties of human glenohumeral cartilage were determined by testing 120 rectangular strips in uniaxial tension and 70 cylindrical plugs in confined compression, obtained from five human glenohumeral joints. Specimens were harvested from five regions across the articular surface of the humeral head and two regions on the glenoid. Tensile strips were obtained along two orientations, parallel and perpendicular to the split-line directions. Two serial slices through the thickness, corresponding to the superficial and middle zones of the cartilage layers, were prepared from each tensile strip and each compressive plug. The equilibrium tensile modulus and compressive aggregate modulus of cartilage were determined from the uniaxial tensile and confined compression tests, respectively. Significant differences in the tensile moduli were found with depth and orientation relative to the local split-line direction. Articular cartilage of the humeral head was significantly stiffer in tension than that of the glenoid. There were significant differences in the aggregate compressive moduli of articular cartilage between superficial and middle zones in the humeral head. Furthermore, tensile and compressive stress-strain responses exhibited nonlinearity under finite strain, while the tensile modulus differed by up to two orders of magnitude from the compressive aggregate modulus at 0% strain, demonstrating a high degree of tension-compression nonlinearity. The complexity of the mechanical properties of human glenohumeral cartilage was exposed in this study, showing anisotropy, inhomogeneity, and tension-compression nonlinearity within the same joint. The observed differences in the tensile properties of human glenohumeral cartilage suggest that the glenoid may be more susceptible to cartilage degeneration than the humeral head.

[Vitamin A, E and C serum concentration in patients with Borrelia burgdorferi antibodies--non-symptomatic carriers]. / Stezenie witamin A, E oraz C W surowicy krwi osób posiadajacych przeciwciala przeciwko Borrelia burgdorferi- bez objawów zakazenia--doniesienie wstepne.

To estimate vitamin A, E and C serum concentrations among forestry workers showing antibodies against Borrelia burgdorferi presence. Vitamins A, E and C concentrations were evaluated in 117 sera of forestry workers. 78 persons aged 18-63 (x=43.07) showed antibodies against Borrelia burgdorferi presence. In this group 13 persons showed presence of IgM, 42 persons with IgG and 23 with IgM and IgG. Control group consisted of 39 persons aged 18-56 years (x=40,97), with no detectable anti-Borrelia burgdorferi antibodies in serum. Serologic diagnosis was performed with use of ELISA kit - Borrelia recombinant IgM, IgG (Biomedica, Austria). Vitamins A and E serum concentrations were detected by RP-HPLC method with spectrophotometric detection (De Leenheet and co.). Vitamin C was detected by RP-HPLS method with spectrophotometric method (Ivanovic and co). Obtains results were statistically analysed. Significantly lower of vitamin A and E serum concentration of persons with anti-borrelia antibodies presence. The lowest concentration was observed in group showing presence of IgM and IgG. No significant difference in vitamin C serum concentration in examined groups was observed. These results may suggest that low serum concentrations of vitamin A and E may have influence on Borrelia burgdorferi infection development.

Direct fluorometric measurement of hepatitis C virus helicase activity.

The non-structural protein 3 (NS3) of hepatitis C virus (HCV) is a highly promising target for anti-HCV therapy because of its multiple enzymatic activities, such as RNA-stimulated nucleoside triphosphatase, RNA helicase and serine protease. The helicase domain of NS3 as well as domain 2 of the helicase were expressed in a baculovirus system to obtain in high yield active proteins for prospective studies of complexes of the helicase with its inhibitors. A novel direct fluorometric test of helicase activity with a quenched DNA substrate, 3' labeled with a Cy3 dye and 5' labeled with a Black Hole Quencher, was developed and optimal reaction conditions established. This test based on fluorescence resonance energy transfer is simple and fast. It allows for direct measurements of enzyme activity, circumventing laborious and complicated radioactive techniques that are poorly reproducible. The results obtained encourage us to propose this new fluorescent assay as a method enabling high throughput screening of anti-helicase compounds.

Effects of amifostine on liver oxidative stress caused by cyclophosphamide administration to rats.

Cyclophosphamide is an inactive cytostatic, which is metabolised into active metabolites mainly in the liver. During bioactivation, reactive oxygen species (ROS) are also formed, which can modify the components of both healthy and neoplastic cells leading to decreased antioxidative capacity. Amifostine is a drug that can inactivate ROS. The aim of the present study was to evaluate the influence of amifostine on the antioxidative system of the liver of rats exposed to cyclophosphamide. Intraperitoneal administration of cyclophosphamide was found to decrease the activity of liver antioxidative enzymes, i.e. superoxide dismutase, glutathione peroxidase and glutathione reductase, and to increase catalase activity. Amifostine slightly influenced antioxidative enzyme activity, causing a significant increase only in superoxide dismutase activity. Co-administration of cyclophosphamide and amifostine nearly prevented changes in activities of superoxide dismutase, glutathione reductase and catalase, as well as to a high degree of glutathione peroxidase. Cyclophosphamide also evoked a decrease in the level of non-enzymatic antioxidants, such as reduced glutathione and vitamins C, E and A, as well as total antioxidant status. Administration of amifostine alone caused a significant increase in non-enzymatic antioxidant level that resulted in an increase in total antioxidant status. Administration of amifostine together with cyclophosphamide to a large extent prevented changes in the evaluated non-enzymatic antioxidative parameters, decreasing values of their concentration to the values of control group. Changes of liver antioxidative abilities during detoxification of cyclophosphamide were accompanied by intensified lipid peroxidation, manifested by an increase in concentration of products such as malondialdehyde and 4-hydroxynonenal. Amifostine caused the inhibition of lipid peroxidation in the liver of both control and cyclophosphamide-treated rats. In conclusion, our results suggest that amifostine significantly protects liver antioxidant properties from changes caused by cyclophosphamide treatment and in consequence prevents oxidative stress and phospholipid peroxidative damage.

Protection against cyclophosphamide-induced renal oxidative stress by amifostine: the role of antioxidative mechanisms.

Cyclophosphamide, an alkylating compound used in chemotheraphy, is metabolized into active metabolites that form reactive oxygen species. Reactive oxygen species can modify the components of both healthy and neoplastic cells in circumstances of decreased antioxidative abilities. That leads to the dysfunction of organs, including the kidneys. Therefore, drugs like amifostine, which protect healthy cells against reactive oxygen species, may be applied during cyclophosphamide therapy. The aim of this study was to evaluate the influence of amifostine on the antioxidative system of the kidneys of rats that were exposed to cyclophosphamide. Intraperitoneal administration of cyclophosphamide was found to decrease the activity of the kidney's antioxidative enzymes, such as superoxide dismutase, glutathione peroxidase, gluthatione reductase, and catalase. Amifostine, however, caused an increase in the activity of these enzymes. The administration of amifostine with cyclophosphamide partially prevented changes in the activities of the examined enzymes observed after cyclophosphamide injection. Cyclophosphamide also evoked a decrease in the levels of nonenzymatic antioxidants, such as reduced gluthatione, vitamin C, and vitamin E as well as the total antioxidant status. The administration of amifostine together with cyclophosphamide prevented changes in the concentration of evaluated nonenzymatic antioxidative parameters, increasing values of their concentration to the values in the control group. Changes in the kidneys' antioxidative abilities during detoxification from cyclophosphamide were accompanied by intensified lipid peroxidation, which was manifested by an increase in the concentration of malondialdehyde and 4-hydroxynonenal. Amifostine caused the inhibition of lipid peroxidation in the kidneys of control and cyclophosphamide-treated rats. In conclusion, our results suggest that amifostine significantly protects kidney antioxidant parameters from changes caused by cyclophosphamide treatment and, in consequence, prevents oxidative stress and phospholipid peroxidative damage. Thus, amifostine prevents renal injury and dysfunction.

[Evaluation of oxidoreductive potential of patients with neuroborreliosis]. / Ocena potencjalu oksydoredukcyjnego u chorych z neuroborelioza.

AIM: The purpose of the study was to evaluate parameters of oxidoreductive system in serum and cerebrospinal fluid (CSF) of patients with neuroborreliosis. MATERIAL AND METHODS: The cases were 25 patients aged 21 to 64 (x = 42.3) hospitalized with diagnosis of neuroborreliosis. Activity of superoxide dismutase (Cu, Zn-SOD), glutathione reductase (GSSG-R), glutathione peroxidase (GSH-Px) and concentration of sulphydryl groups (-SH) and malondialdehyde (MDA) in serum and CSF were tested. The control group consisted of 10 patients with diagnosis of discopathy. An examination was performed twice: before and after treatment. RESULTS: Results of the study showed lack of stability in an oxidoreductive system during neuroborreliosis both in serum and in CSF. In CSF activity of SOD was increased while activity of GSH-Px and GSSG-R were decreased. Also concentration of -SH and lipid peroxidation products measured as MDA were increased. The increase of SOD, GSH-Px, GSSG-R activity and concentration of -SH and MDA in serum were detected. CONCLUSIONS: Disorders of an oxidoreductive system in CSF and serum during neuroborreliosis were observed. These changes persisted despite treatment and normalization of inflammatory CSF markers.