PAH metabolites in European eels (Anguilla anguilla) as indicators of PAH exposure: different methodological approaches.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous contaminants of aquatic environments derived from pyrogenic and petrogenic sources. In fish, as in other vertebrates, PAHs are rapidly metabolized. However, the metabolites have been proven to induce multiple deleterious effects in fish. The concentrations of biliary polycyclic aromatic hydrocarbon metabolites in eels (Anguilla anguilla) caught in Polish waters were measured. The main objectives of the study were to provide information on the levels of PAH metabolites in eels inhabiting Polish waters and to discuss which methodological approach is appropriate for assessing PAH exposure in aquatic ecosystems. The non-normalized median concentration of 1-OH Pyr and 1-OH Phe measured in eels from Polish waters ranged from 11 to 1642 ng ml(-1) bile and from 83 to 929 ng ml(-1) bile, respectively, depending on the sampling site. Data normalization in relation to bile pigment content reduced inter-site variation, and the normalized median concentrations of 1-OH Pyr and 1-OH Phe ranged from 0.44 to 20.24 ng A(-1)380 and from 1.58 to 11.11 ng A(-1)380, respectively. Our study indicated that results were more consistent for the two species sampled in the same area (eel examined in the current study and flounder examined in our previous study) when the fluorescence response of diluted bile samples was compared than when concentrations of 1-OH Pyr determined with the mean of HPLC were compared.

Serum IL-17F does not predict poor response to IM IFNß-1a in relapsing-remitting MS.

OBJECTIVE: There is a significant unmet need for serum biomarkers in relapsing-remitting multiple sclerosis (RRMS) that are predictive of therapeutic response to disease-modifying therapies. Following a recent Stanford study which reported that pretreatment levels of serum interleukin (IL)-17F could predict poor response to interferon-ß (IFNß) therapy, we sought to validate the finding using samples from a large clinical trial. METHODS: The validation cohort included 54 good responders (GR) and 64 poor responders (PR) selected from 762 subjects with RRMS from the IM IFNß-1a dose comparison study (Avonex study C94-805). Subjects were classified as GR and PR based on the number of relapses, Expanded Disability Status Scale score, and new and enlarging T2 lesions on MRI. Serum samples were assayed for IL-17F using a multiplexed Luminex assay and for IL-17F/F using an ELISA. Replicate aliquots from the Stanford study were also assayed to assure reproducibility of methods. RESULTS: Median pretreatment and post-treatment serum IL-17F levels were not statistically significantly different between GR and PR, and serum IL-7/IL-17F ratios were also not predictive of response status. Replicate aliquots from the Stanford study showed good correlation to their original cohort (r = 0.77). CONCLUSIONS: We were unable to validate the finding that serum IL-17F is a predictor of PR in a large independent cohort of subjects with RRMS. Differences in patient populations and methodology might explain the failure to validate the results from the Stanford study.

Assessment of PAH pollution in the southern Baltic Sea through the analysis of sediment, mussels and fish bile.

The concentrations of fifteen PAH compounds in samples of sediment and blue mussel tissue (Mytilus trossulus) were measured. In addition, the biliary polycyclic aromatic hydrocarbon metabolites present in flounder (Platichthys flesus) were analysed. Two methods were used in the analysis of PAH metabolites; high performance liquid chromatography (HPLC) and fixed wavelength fluorescence (FF). The major PAH metabolite which could be measured using the HPLC method was 1-OH pyrene. It was possible to detect 1-OH Phe and 3-OH B[a]P in 70 and 24 samples respectively, of the 87 samples analysed. However, the concentrations of 1-OH Phe and 3-OH B[a]P were below or near to the LOQ (0.002 µg ml(-1) bile). The bile of flounder samples from the Gulf of Gdansk had 1-OH Pyr concentrations which ranged from 0.019 to 0.066 µg ml(-1) bile. The high linear correlation observed between the quantity of 1-OH pyrene determined by the HPLC-F method and the content of the sum of pyrene-type PAHs obtained by the FF method indicated the FF method of determination of pyrene-type PAH metabolites can be used as a screening method. The content of ∑(15)PAHs in sediments collected in the Gulf of Gdansk, in 2008, ranged from 29.3 to 103 µg kg(-1) dw. In mussel tissue ∑(15)PAHs concentrations were between 173.2 µg kg(-1) dw and 237.7 µg kg(-1) dw. All concentrations measured in the current study, in mussel tissue, were below the OSPAR toxicity threshold values.

Short-Term Relapse Quantitation as a Fully Surrogate Endpoint for Long-Term Sustained Progression of Disability in RRMS Patients Treated with Natalizumab.

Time to sustained worsening in the expanded disability status scale as the standard for evaluating the accumulation of disability has been used as a measure of clinical efficacy in many relapsing-remitting multiple sclerosis (RRMS) clinical trials. However, this measurement usually requires a large sample and long-term study to demonstrate the treatment effect. Annualized relapse rate or time to first relapse is also widely used as alternative measurements of clinical efficacy. A formal statistical validation of short-term relapse activity as a surrogate endpoint for long-term sustained progression of disability could potentially permit smaller, shorter, and less expensive clinical trials in RRMS. Four statistical validation/evaluation approaches consistently showed that relapse activity through one year of treatment serves as statistically valid surrogate endpoint for time to sustained progression of disability. The analysis demonstrates that long-term sustained progression of disability can be predicted by short-term relapse measures with 4 consistent validations of statistical approaches, including a formal statistical hypothesis test. This was demonstrated in a large phase III trial of natalizumab and showed that the beneficial clinical effect of natalizumab on sustained progression of disability at 2 years in patients with RRMS can be predicted by the total number of relapses at 1 year.

Variation in fat content and fatty-acid composition of the Baltic herring Clupea harengus membras.

The fat content and fatty-acid profiles of herring, Clupea harengus membras, from the southern Baltic Sea varied depending on when (fishing season) and where (fishing grounds) the fish were caught as well as on their size and sex. The fat, protein and dry matter content and the fatty-acid profiles were assayed in C. h. membras muscle tissue. The changes observed in fatty-acid profiles were determined by factors such as specimen mass and fat content, which, in turn, depended on fishing season. This is explained by dietary differences between juvenile and older fish. Gonad maturation and spawning in the latter are also factors. The study results provide confirmation of the hypothesis that polyunsaturated fatty acids (PUFA), in particular docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), play vital roles in the sexual maturation of C. h. membras.

Proof of concept studies for tissue-protective agents in multiple sclerosis.

BACKGROUND: There is considerable interest in tissue-protective treatments for multiple sclerosis (MS). METHODS AND OBJECTIVES: We convened a group of MS clinical trialists and related researchers to discuss designs for proof of concept studies utilizing currently available data and assessment methods. RESULTS: Our favored design was a randomized, double-blind, parallel-group study of active treatment versus placebo focusing on changes in brain volume from a post-baseline scan (3-6 months after starting treatment) to the final visit 1 year later. Study designs aimed at reducing residual deficits following acute exacerbations are less straightforward, depending greatly on the anticipated rapidity of treatment effect onset. CONCLUSIONS: The next step would be to perform one or more studies of potential tissue-protective agents with these designs in mind, creating the longitudinal data necessary to refine endpoint selection, eligibility criteria, and sample size estimates for future trials.

Differential diagnosis of suspected multiple sclerosis: a consensus approach.

BACKGROUND AND OBJECTIVES: Diagnosis of multiple sclerosis (MS) requires exclusion of diseases that could better explain the clinical and paraclinical findings. A systematic process for exclusion of alternative diagnoses has not been defined. An International Panel of MS experts developed consensus perspectives on MS differential diagnosis. METHODS: Using available literature and consensus, we developed guidelines for MS differential diagnosis, focusing on exclusion of potential MS mimics, diagnosis of common initial isolated clinical syndromes, and differentiating between MS and non-MS idiopathic inflammatory demyelinating diseases. RESULTS: We present recommendations for 1) clinical and paraclinical red flags suggesting alternative diagnoses to MS; 2) more precise definition of "clinically isolated syndromes" (CIS), often the first presentations of MS or its alternatives; 3) algorithms for diagnosis of three common CISs related to MS in the optic nerves, brainstem, and spinal cord; and 4) a classification scheme and diagnosis criteria for idiopathic inflammatory demyelinating disorders of the central nervous system. CONCLUSIONS: Differential diagnosis leading to MS or alternatives is complex and a strong evidence base is lacking. Consensus-determined guidelines provide a practical path for diagnosis and will be useful for the non-MS specialist neurologist. Recommendations are made for future research to validate and support these guidelines. Guidance on the differential diagnosis process when MS is under consideration will enhance diagnostic accuracy and precision.

Ethics of placebo-controlled clinical trials in multiple sclerosis: a reassessment.

The increasing number of established effective therapies for relapsing multiple sclerosis (MS) and emerging consensus for early treatment raise practical concerns and ethical dilemmas for placebo-controlled clinical trials in this disease. An international group of clinicians, ethicists, statisticians, regulators, and representatives from the pharmaceutical industry convened to reconsider prior recommendations regarding the ethics of placebo-controlled trials in MS. The group concluded that placebo-controlled trials can still be done ethically, with restrictions. For patients with relapsing MS for which established effective therapies exist, placebo-controlled trials should only be offered with rigorous informed consent if the subjects refuse to use these treatments, have not responded to them, or if these treatments are not available to them for other reasons (e.g., economics). Suggestions are provided to protect subject autonomy and improve informed consent procedures. Recommendations are tighter than previously suggested for placebo-controlled trials in "resource-restricted" environments where established therapies may not be available. Guidance is also provided on the ethics of alternative trial designs and the balance between study subject burden and risk, scientific rationale and interpretability of trial outcomes.

Transient backbending behavior in the Ising model with fixed magnetization.

The physical origin of the backbendings in the equations of state of finite but not necessarily small systems is studied in the Ising model with fixed magnetization (IMFM) by means of the topological properties of the observable distributions and the analysis of the largest cluster with increasing lattice size. Looking at the convexity anomalies of the IMFM thermodynamic potential, it is shown that the order of the transition at the thermodynamic limit can be recognized in finite systems independently of the lattice size. General statistical mechanics arguments and analytical calculations suggest that the backbending in the caloric curve is a transient behavior which should not converge to a plateau in the thermodynamic limit, while the first-order transition (in the Ehrenfest sense) is still signaled by a discontinuity in the magnetization equation of state.

Dynamics of many-particle fragmentation in a cellular automaton model.

A three-dimensional cellular automaton model developed by the authors to deal with the dynamics of N-body interactions has been adapted to investigate the head-on collision of two identical bound clusters of particles, and the ensuing process of fragmentation. The range of impact energies is chosen low enough, to secure that a compound bound cluster can be formed. The model is devised to simulate the laboratory set-up of fragmentation experiments as monitored by 4 pi detectors. The particles interact via a Lennard-Jones potential. At low impact energies the numerical experiments following the dynamics of the individual particles indicate a phase of energy sharing among all the particles of the compound cluster. Fragments of all sizes are then found to evaporate from the latter cluster. The cluster sizes, measured in our setup by simulated 4 pi detectors, conform to a power law of exponent approximately 2.6. In an attempt to duplicate the laboratory caloric curves related, in particular, to nuclear fragmentation processes, we introduce several temperature parameters (kinetic temperature of nucleons, kinetic temperature of fragments, reaction equilibrium temperatures). Theoretical caloric curves are then constructed for those temperature parameters, we regard as physically most relevant. Our results show that different temperature definitions generate different curve patterns, indicating that the fragmentation system remains far from thermodynamic equilibrium. The pattern of the laboratory caloric curve for Au-Au collision experiments as derived from a recent analysis [NuPECC Report, 1997 (unpublished)] is reproduced qualitatively by our reaction temperatures.

MFAME, N-methyl-N-D-fructosyl amphotericin B methyl ester, a new amphotericin B derivative of low toxicity: relationship between self-association and effects on red blood cells.

In aqueous solutions N-methyl-N-D-fructosyl amphotericin B methyl ester (MFAME), a novel amphotericin B derivative with low animal toxicity, similar to its parent antibiotic, exists in three forms: monomeric, soluble and insoluble aggregates in equilibrium [1]. The aim of our work was to examine the influence of medium composition on the MFAME self-association and the relationship between MFAME self-association and its toxicity towards red blood cells. The toxicity of MFAME in aggregated state towards red blood cells was tested by measuring the induction of potassium leakage and extent of haemolysis. The proportions of antibiotic species present in various aqueous media were determined by analysis of the UV-Vis spectra as a function of the antibiotic concentration. Numeric decomposition of the spectra allowed identification of four spectral species present in MFAME solutions: monomeric and three aggregated forms. Our results indicate that these aggregates, named type I, type II and type III, are different in terms of spectral properties, as well as effectiveness towards red blood cells. Soluble aggregate types I and III are the active forms of MFAME towards erythrocytes. The medium composition seems to be the main factor determining which type of antibiotic aggregate prevails in solution.

Demyelination occurring during anti-tumor necrosis factor alpha therapy for inflammatory arthritides.

OBJECTIVE: To review the occurrence of neurologic events suggestive of demyelination during anti-tumor necrosis factor alpha (anti-TNFalpha) therapy for inflammatory arthritides. METHODS: The Adverse Events Reporting System of the Food and Drug Administration (FDA) was queried following a report of a patient with refractory rheumatoid arthritis who developed confusion and difficulty with walking after receiving etanercept for 4 months. RESULTS: Nineteen patients with similar neurologic events were identified from the FDA database, 17 following etanercept administration and 2 following infliximab administration for inflammatory arthritis. All neurologic events were temporally related to anti-TNFalpha therapy, with partial or complete resolution on discontinuation. One patient exhibited a positive rechallenge phenomenon. CONCLUSION: Further surveillance and studies are required to better define risk factors for and frequency of adverse events and their relationship to anti-TNFalpha therapies. Until more long-term safety data are available, consideration should be given to avoiding anti-TNFalpha therapy in patients with preexisting multiple sclerosis and to discontinuing anti-TNFalpha therapy immediately when new neurologic signs and symptoms occur, pending an appropriate evaluation.

Neuropsychological effects of interferon beta-1a in relapsing multiple sclerosis. Multiple Sclerosis Collaborative Research Group.

Cognitive dysfunction is common in multiple sclerosis (MS), yet few studies have examined effects of treatment on neuropsychological (NP) performance. To evaluate the effects of interferon beta-1a (IFNbeta-1a, 30 microg administered intramuscularly once weekly [Avonex]) on cognitive function, a Comprehensive NP Battery was administered at baseline and week 104 to relapsing MS patients in the phase III study, 166 of whom completed both assessments. A Brief NP Battery was also administered at 6-month intervals. The primary NP outcome measure was 2-year change on the Comprehensive NP Battery, grouped into domains of information processing and learning/memory (set A), visuospatial abilities and problem solving (set B), and verbal abilities and attention span (set C). NP effects were most pronounced in cognitive domains vulnerable to MS: IFNbeta-1a had a significant beneficial effect on the set A composite, with a favorable trend evident on set B. Secondary outcome analyses revealed significant between-group differences in slopes for Brief NP Battery performance and time to sustained deterioration in a Paced Auditory Serial Addition Test processing rate, favoring the IFNbeta-1a group. These results support and extend previous observations of significant beneficial effects of IFNbeta-1a for relapsing MS.

Comparative studies on cell stimulatory, permeabilizing and toxic effects induced in sensitive and multidrug resistant fungal strains by amphotericin B (AMB) and N-methyl-N-D-fructosyl amphotericin B methyl ester (MFAME).

N-Methyl-N-D-fructosyl-amphotericin B methyl ester (MFAME) is a new derivative of amphotericin B, which is characterised by low toxicity to mammalian cells and good solubility in water of its salts. The antifungal activity and effects of MFAME towards Candida albicans and Saccharomyces cerevisiae multidrug resistant MDR(+) and sensitive MDR(-) strains was compared with those of parent compound. The results obtained indicate that MDR(+) S. cerevisiae was sensitive to MFAME as well as to AMB. MFAME exhibited the same effects on fungal cells studied as parent antibiotic. The two antibiotics, depending on the dose applied induced cell stimulation, K+ efflux, and/or had a toxic effect.

A longitudinal study of T1 hypointense lesions in relapsing MS: MSCRG trial of interferon beta-1a. Multiple Sclerosis Collaborative Research Group.

BACKGROUND: T1 hypointense lesions (T1 black holes) are focal areas of relatively severe CNS tissue damage detected by MRI in patients with MS. OBJECTIVE: To determine the natural history of T1 hypointense lesions in relapsing MS and the utility of T1 hypointense lesions as outcome measures in MS clinical trials. METHODS: MR studies were from the Multiple Sclerosis Collaborative Research Group trial. Longitudinal results are reported in 80 placebo- and 80 interferon beta-1a (IFNbeta-1a)-treated patients with mild to moderate disability relapsing-remitting MS. RESULTS: There was a small but significant correlation between T1 hypointense lesion volume and disability at baseline and on trial (r = 0.22, r = 0.28). In placebo patients there was a 29.2% increase in the mean volume of T1 hypointense lesions (median 124.5 mm3) over 2 years (p < 0.001 for change from baseline), as compared to an 11.8% increase (median 40 mm3) in the IFNbeta-1a-treated patients (change from baseline not significant). These treatment group comparisons did not quite reach significance. The most significant contributor to change in T1 hypointense lesions was the baseline number of enhancing lesions (model r2 = 0.554). Placebo patients with more active disease, defined by enhancing lesions at baseline, were the only group to show a significant increase in T1 hypointense lesion volume from baseline. CONCLUSION: The development of T1 hypointense lesions is strongly influenced by prior inflammatory disease activity, as indicated by enhancing lesions. These results suggest that treatment with once weekly IM IFNbeta-1a (30 mcg) slows the 2-year accumulation of these lesions in the brain.

EAE susceptibility in FVB mice.

The in vivo study of immunologic mechanisms in experimental allergic encephalomyelitis (EAE) requires, in some instances, the use of transgenic mice. As FVB mice represent a strain whose fertilized ova possess a relatively large pronucleus, this is a preferred strain in which to generate mice that are transgenic for a variety of genes. An important issue, then, is whether the FVB background supports the development of EAE. We report here that relapsing EAE is easily and reproducibly induced in FVB mice with whole myelin basic protein in complete Freund's adjuvant.

Localization of the human SP3 gene to chromosome 7p14-p15.2. The lack of expression in multiple sclerosis does not reflect abnormal gene organization.

Sp3 belongs to a large family of transcription factors that bind GC/T box elements. We have previously demonstrated the deficient expression of Sp3 in peripheral blood mononuclear cells (PBMC) from most patients with multiple sclerosis (MS). In the current study, the Sp3 gene was assigned to chromosome 7 by using somatic cell hybrid mapping and analysis of a chromosome 7-specific cosmid library. The position of Sp3 was refined to 7p14-p15.2 by fluorescence in situ hybridization (FISH). Southern blot and polymerase chain reaction analysis of genomic DNA failed to demonstrate a detectable difference between MS and control PBMC.