Ms

Keywords: HIV-1 protease inhibitors; structure-based drug design; pyrrolidine-based inhibitors; crystallography; deviating binding modes Structure-based drug design is an integral part of industrial and academic drug discovery projects. Initial lead structures are, in general, optimized in terms of affinity using iterative cycles comprising synthesis, biological evaluation, computational methods, and structural analysis. X-ray crystallography commonly suggests the existence of a single well-defined state, termed binding mode, which is generally assumed to be consistent in a series of similar ligands and therefore used for the following optimization process. During the further development of symmetrically disubstituted 3,4-amino-pyrrolidines as human immunodeficiency virus type 1 protease inhibitors, we discovered that, by modification of the P1/P1′ moieties of our lead structure, the activity of the inhibitors towards the active-site mutation Ile84Val was altered, however, not being explainable with the initial underlying structure-activity relationship. The cocrystallization of the most potent derivative in complex with the human immunodeficiency virus type 1 protease surprisingly led to two different crystal forms (P2 1 2 1 2 1 and P6 1 22). Structural analysis revealed two completely different binding modes; the interaction of the pyrrolidine nitrogen atom with the catalytic aspartates remains as the only similarity. The study presented clearly demonstrates that structural biology has to escort the entire lead optimization process not to fail by an initially observed binding orientation.

Objective Multiple Sclerosis (MS) is known as one of the most common neurological disorders that affect patients' lives tremendously. These people experience various dysfunctions. One of the most common problems in patients with MS is a balance deficit, which limits the participation and increases the risk of falling. Recently various studies were focused on finding the causes of this imbalance. On the other hand, these patients experience cognitive problems, such as attention deficit and cognitive flexibility. Therefore, the aim of this study was to investigate the relationship between cognitive flexibility and selective attention with balance in patients with relapsing-remitting MS. Materials & Methods This study is a cross-sectional case study. The statistical population in this study was patients with MS who referred to Kashani Hospital and Alzahra Isfahan Hospital in 1395 for inclusion in the study, based on available sampling method. MS diagnosis was performed in these centers with the approval of at least one adult neurologist. Finally, 40 patients with MS were selected by convenient sampling. A written consent form was received from all the families and the patients with MS themselves for participation in the study. All patients were kept confidential in the study, and they were allowed to leave during the study if they refused. The inclusion criteria to participate in the study were: satisfaction of patients to participate in the plan, the individual received a diagnosis of relapsing-remitting MS from those who were referred to Al-Zahra and Kashani hospitals in Isfahan, aged 20 to 40 years, at least two attacks in last 24 months, a score of less than 5.5 in EDSS was associated with a moderate degree of MS due to the severity of the disability and the symptoms of the disease, having sufficient insight and the ability to collaborate in the study through the MMSE test. The exclusion criteria of the study were: dissatisfaction of the patients with the continuation of the study, presence and participation in similar investigations in the last three months that were performed intermittently on executive functions, the presence of visual and auditory impairment, recurrence and severity of MS, disability from moderate to severe, and any abnormalities such as seizure. After that test EDSS, Stroop color-word test, Wisconsin card sorting, and Berg Balance Scale were completed by patients with MS. The results of the tests of executive function and balance were analyzed using SPSS20 software for the presence or absence of the relationship between executive function is determined by the balance. To analyze the data, Pearson correlation coefficient and multiple regression and analysis of variance with repeated measures at the level of 0.05. Results The results of this study demonstrate a significant relationship between selective attention and balance problem (P<0.05). However, there is no significant relationship between cognitive flexibility and balance (P<0.05). As well as the selective attention can predict the balance problem (P<0.05) (P<0.01). Conclusion A significant relationship between selective attentions to balance demonstrates that appropriately designed interventions to repair the attention, can decrease the balance problem in patients with MS. One of the possible reasons for this relationship can be found in the brain system that attention circuits are interacting with equilibrium systems, and any defect in selective attention leads to a loss of balance. Patients with MS experience constant imbalance due to cerebral plaques and relapsing-remitting periods, and in addition to rehabilitation of the balance system, cognitive systems such as attention should be treated.

We have evaluated the effects of the proline homologue Aze (1) (azetidine-2-carboxylic acid) on growth of Escherichia coli strains used to over-express recombinant forms of murine myelin basic protein (rmMBP), and on the degree of misincorporation. Addition of Aze to minimal media resulted in severe diminution of growth rate, but rmMBP could still be produced and purified. Mass spectrometry indicated that a detectable proportion of the rmMBP produced had incorporated Aze instead of proline (Pro), to a maximum of three of eleven possible sites. Molecular modelling of a proline-rich region of rmMBP illustrated that the misincorporation of Aze at any site would cause a severe bend in the polypeptide chain, and that multiple Pro ? Aze substitutions would completely disrupt a poly-proline type II structure that has been conjectured to be functionally significant.

Restless legs syndrome (RLS) is a neurological disorder that may be related to iron misregulation at the level of the central nervous system. Evidence that iron is involved in RLS comes from magnetic resonance imaging data, autopsy studies, analyses of cerebrospinal fluid (CSF), and correlations of symptoms with serum ferritin. To further examine the possibility that brain iron status is insufficient in RLS, we determined ferritin levels in the CSF. Specifically, we differentiated between the H-and L-subunits of ferritin, because these peptides are expressed from different chromosomes and have different functions. We measured H-and L-ferritin subunit levels in control and RLS human CSF using immunoblot analysis and found that both H-and L-ferritin are significantly decreased in early but not late-onset RLS. Additionally, we quantified total protein in each CSF sample to establish that the decrease in ferritin subunits in RLS did not reflect a decrease in total protein in CSF. Furthermore, we used equal amounts of total CSF protein in the immunoblot analyses, in contrast to previously published studies that provided only volumetric data, to determine which approach was more accurate for quantifying the amount of ferritin relative to other proteins in CSF. Our results establish a protein standard in RLS, provide a comparative analysis of protein-controlled versus volumetric immunoblot techniques, and argue for a profound loss of iron storage capacity in the brain in RLS, specifically in the early onset RLS phenotype. These data suggest that CSF ferritin levels may provide a biomarker for assisting in the diagnosis of RLS. (J Lab Clin Med 2006;147:67-73)

CITATIONS 11 READS 112 4 authors: Some of the authors of this publication are also working on these related projects: Biomass pyrolysis, gasification, catalytic pyrolysis, catalytic upgrading of pyrolysis oil, Technoeconomic analysis of biomass pyrolysis process, Carbon capture and storage View project CO2 reforming of methane over Ni/MgO Catalysts Promoted with Zr and La Oxides View project

We have previously described methodology that makes it possible to genetically encode a wide array of unnatural amino acids in both prokaryotic and eukaryotic organisms. Here, we report the systematic optimization of a Saccharomyces cerevisiae expression system for the production of mutant proteins containing unnatural amino acids. Modifications include significant increases in both the expression levels of the orthogonal Escherichia coli amber suppressor tRNA CUA and cognate aminoacyl-tRNA synthetase (aaRS) pair, and expression of the target protein gene using a strong transcriptional promoter, optimized codons and elevated plasmid copy numbers. With this new system, a number of unnatural amino acids, including the photocrosslinkers p-benzoylphenylalanine and p-azidophenylalanine, and the chemically reactive amino acids, p-acetylphenylalanine and p-propargyloxyphenylalanine, were incorporated into human superoxide dismutase (hSOD) in yeast in good yields (maximally ∼6-8 mg/l of culture in most cases). Mass spectrometric analysis of the hSOD mutants was performed with high dynamic range using multiple reaction monitoring that provided new insights into the factors that control the fidelity of unnatural amino acid incorporation.

Genes for functional Ser/Thr protein kinases (STPKs) are ubiquitous in prokaryotic genomes, but little is known about their physiological substrates and their actual involvement in bacterial signal transduction pathways. We report here the identification of GarA (Rv1827), a Forkheadassociated (FHA) domain-containing protein, as a putative physiological substrate of PknB, an essential Ser/Thr protein kinase from Mycobacterium tuberculosis. Using a global proteomic approach, GarA was found to be the best detectable substrate of the PknB catalytic domain in non-denatured whole-cell protein extracts from M. tuberculosis and the saprophyte Mycobacterium smegmatis. Enzymological and binding studies of the recombinant proteins demonstrate that docking interactions between the activation loop of PknB and the C-terminal FHA domain of GarA are required to enable efficient phosphorylation at a single N-terminal threonine residue, Thr22, of the substrate. The predicted amino acid sequence of the garA gene, including both the N-terminal phosphorylation motif and the FHA domain, is strongly conserved in mycobacteria and other related actinomycetes, suggesting a functional role of GarA in putative STPK-mediated signal transduction pathways. The ensuing model of PknB-GarA interactions suggests a substrate recruitment mechanism that might apply to other mycobacterial kinases bearing multiple phosphorylation sites in their activation loops.

Tepoxalin is a veterinary drug registered for use in the dog as a dual inhibitor (cyclooxygenase -5 lipoxygenase). In the horse, it predominantly triggers a strong cyclooxygenase inhibition; this bias seems to be due to the action of its metabolite(s). Among these, only the RWJ-20142 is well known, while to the best of our knowledge no information is available on the other metabolites produced in vivo. Hence, the identification of its main metabolic pathway is pivotal to better understand its clinical activity.

People make decisions every day. There are too many choices and a little time to explore them all. Recommendation systems help people make decisions in these complex information spaces. Recommendation systems are a type of information filtering that presents lists of items (films, songs, books, videos, images, products, web pages) which are likely of user interest. Amazon, Last.fm, Ulike, iLike, Netflix, Pandora are the most popular recommender systems all over the world. Simply they compare user interest acquired from his/her profile with some reference characteristics and predict the rating that the user would give. Recommendation System is a subclass of information filtering system which takes input from users and provides User with the most suitable output to fulfill his requirements. This system is used to present a new college admission system using data mining techniques for tackling college admission prediction problems. This System uses content-based filtering to provide aspiring students (Master of Science) with the most appropriate choices of colleges based on different parameters. The system analyses the student academics, merit, background, student records and the college admission criteria. Then it predicts the likelihood of colleges the student may enter. In addition to the high prediction accuracy is an advantage, as the system can predict suitable colleges that match the students' profiles and the suitable track channels through which the students are advised to enter.

This review addresses the use of high-performance liquid chromatography (HPLC) and capillary electrophoresis (CE) as affinity separation methods to characterise drugs or potential drugs-bio-polymer interactions. Targets for the development of new drugs such as enzymes (IMERs), receptors, and membrane proteins were immobilized on solid supports. After the insertion in the HPLC system, these immobilized bio-polymers were used for the determination of binding constants of specific ligands, substrates and inhibitors of pharmaceutical interest, by frontal analyses and zonal elution methods. The most used bio-polymer immobilization techniques and methods for assessing the amount of active immobilized protein are reported. Examples of increased stability of immobilized enzymes with reduced amount of used protein were shown and the advantages in terms of recovery for reuse, reproducibility and on-line high-throughput screening for potential ligands are evidenced. Dealing with the acquisition of relevant pharmacokinetic data, examples concerning human serum albumin binding studies are reviewed. In particular, papers are reported in which the serum carrier has been studied to monitor the enantioselective binding of chiral drugs and the mutual interaction between co-administered drugs by CE and HPLC. Finally CE, as merging techniques with very promising and interesting application of microscale analysis of drugs' binding parameters to immobilized bio-polymers is examined.

In this review, we summarize the most recent analytical developments aimed at employing Ionic liquids (ILs) in dispersive liquid-liquid microextraction (DLLME). Four main operation modes can be distinguished: (1) conventional IL-DLLME; (2) temperature-controlled IL-DLLME; (3a) ultrasound-assisted, (3b) microwave-assisted or (3c) vortex-assisted IL-DLLME; and, (4) in-situ IL-DLLME. In these modes, the dispersive solvent can be an organic solvent, a surfactant, or a hydrophilic IL. In some cases, a dispersive solvent is not even necessary. We discuss practical applications of IL-DLLME to determine metals and organic compounds in a variety of samples.

In this study, the N-methylpolyvinylpyridinuim polymer has for the first time been used as a silica surface modifier for CE in combination with ESI MS (CE-ESI-MS). The compatibility for ESI-MS was demonstrated by the analysis of peptides and protein digests. The N-methylpolyvinylpyridium surface interacts electrostatically with the ionized silanol groups, giving a cationic surface with a reversed EOF. The surface modifier gave rapid and repeatable separations of peptides, proteins and protein digests at acidic pH for more than 4 h of continuous use. The CE separation yielded peak efficiencies of up to 4.3 Â 10 5 plates/m. The surface coating is highly compatible with ESI and facilitates the separation and analysis of complex peptide mixtures as shown by the analysis of BSA digests.

A high copy number, selectable marker gene (SMG)-free Agrobacterium binary vector pBin19 nptII was constructed by deleting the nptII gene from pBin19. The binary vectors with the RK2 and pVS replication origins exist in 12 and 3 copies, respectively, in Agrobacterium. The tobacco osmotin gene (ap24) was cloned in pBin19 nptII and the resultant plasmid pBin19 nptII-ap24 was mobilized into the Agrobacterium tumefaciens strain C58C1 Rif r harbouring the single-copy cointegrate vector pGV2260::pSSJ1. The T-DNA of the cointegrate vector harboured the hph (SMG) and gus genes. Transformation of Oryza sativa L. var. Pusa Basmati1 with Agrobacterium tumefaciens (pGV2260::pSSJ1, pBin19 nptII-ap24) yielded 14 independent hyg + /GUS + transgenic plants. Southern blot analysis with hph and ap24 probes revealed that 12 out of the 14 transgenic plants were co-transformed and harboured hph, gus and ap24 genes. The new multi-copy binary vector yielded 86% co-transformation efficiency. SMG elimination by genetic separation of the cointegrate T-DNA with the hph/gus genes and binary vector T-DNA with the ap24 gene was accomplished in four out of ten primary co-transformants that were forwarded to the T 1 generation.

Objective To determine prevalence of metabolic syndrome (MS) among adolescents by using definitions from the National Cholesterol Education Program Adult Treatment Panel III (NCEP) and World Health Organization (WHO) guidelines and to compare the populations identified by these definitions.

Current cancer research is being increasingly focused on the study of distinctive characters of tumour metabolism, resulting in a switch from oxidative phosphorylation to glycolysis (Warburg effect). Isoform 5 of human lactate dehydrogenase (hLDH5), which catalyzes the final step in the glycolytic cascade (pyruvate to lactate), constitutes a relatively new and untapped anti-cancer target. In this study, careful design and synthesis of a selected series of aryl-substituted N-hydroxyindole-2-carboxylates (NHIs) has led to several hLDH5-inhibitors, showing "first-in-class" potency and isoform selectivity. Enzyme kinetics studies indicated that these inhibitors exhibit a competitive mode of inhibition. Some representative examples were tested against two human pancreatic carcinoma cell lines, and displayed a good antiproliferative activity, which was even more evident under hypoxic conditions.

Amitrityline and its metabolite nortriptiline are tricyclic antidepressant drugs widely used for the treatment of several psychiatric disorders. Several methods have been published for the determination of these two antidepressant drugs in pharmaceuticals, biological materials and environmental samples. In this review some of analytical techniques such as ultraviolet/visible spectrophotometry, fluorimetry, capillary electrophoresis, and chromatographic methods (gaschromatography and high-performance liquid chromatography) were discussed. Although HPLC and capillary electrophoresis methods are extensively employed in spite of that UV/VIS spectrophotometry are still popular because of the inherent simplicity, low cost, and reliability for determination of drugs in pharmaceutical preparations.

There is a need for new approaches for enhancing education for sustainable development in universities. Memetics, which is about effective pathways of communication, could be such a new, promising approach. Quality assurance is required in universities to secure and improve education, which could be another approach. The aim of this study is to look into whether and how frameworks for processes and procedures for quality assurance, such as management systems, could be utilized to promote higher education for sustainable development. The study approaches this from both a theoretical and a practical standpoint. An evolutionary perspective was chosen, considering higher education for sustainable development and management systems as memes, or basic units of cultural replication. The practical context was studied by looking into how 11 universities in the Nordic countries have enhanced ESD with management systems. We found that both higher education for sustainable development and management systems could be considered successful memes and that management systems could be applied to enhance higher education for sustainable development.

A simple, sensitive, selective, and rapid high-performance liquid chromatography-tandem mass spectrometry method is developed and validated for the quantitation of naratriptan, using sumatriptan as internal standard (IS). The method included liquid-liquid extraction of naratriptan and IS with methyl-tert-butyl ether and dichloromethane mixture from 100 µL human plasma. The chromatographic separation is achieved on ACE C18 (50 mm × 2.1 mm, 5 µm) analytical column under isocratic conditions, using 0.1% acetic acid and acetonitrile (15:85, v/v) at a flow-rate of 0.4 mL/min. The precursor → product ion transitions for naratriptan (m/z 336.10 → 98.06) and IS (m/z 296.09 → 251.06) were monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring (MRM) and positive ion mode. The linearity of the method for naratriptan is determined in the range of 103-20690 pg/mL with the analysis time of 1.5 min. The method is fully validated according to USFDA guidelines. A systematic post-column infusion study is conducted for ionsuppression due to endogenous matrix components. The process efficiency of analyte (96%) and IS (93%) from spiked plasma samples was consistent and reproducible. The application of the method is demonstrated by a bioequivalence study of 2.5 mg naratriptan tablet formulation in 31 healthy volunteers under fasting conditions.

A B S T R A C T Shoot tip explants obtained from in vitro proliferated shoots were encapsulated in 3% sodium alginate and 100 mM calcium chloride for the production of synthetic seed in Solanum nigrum L., a medicinally important plant. Morphogenic responses of encapsulated shoot tips to various sowing media (full or half-strength 0.8% agar-solidified or liquid MS medium or full-strength MS medium containing BAP) were evaluated in vitro. Of the six media evaluated, maximum conversion was obtained on 0.8% agarsolidified growth regulator free full-strength MS medium. The addition of MS nutrients in alginate matrix had a pronounced effect on the length of shoots that emerged from alginate beads. Encapsulated shoot tips also converted when directly sown in sterile soil moistened with liquid MS medium. Encapsulated shoot tips could be stored at low temperature (4 8C) up to 60 days. Plantlets regenerated from encapsulated shoot tips were acclimatized successfully. ß

A B S T R A C T Peptide and protein (P/P) drugs have been identified as showing great promises for the treatment of various neurodegenerative diseases. A major challenge in this regard, however, is the delivery of P/P drugs over the blood-brain barrier (BBB). Intense research over the last 25 years has enabled a better understanding of the cellular and molecular transport mechanisms at the BBB, and several strategies for enhanced P/P drug delivery over the BBB have been developed and tested in preclinical and clinicalexperimental research. Among them, technology-based approaches (comprising functionalized nanocarriers and liposomes) and pharmacological strategies (such as the use of carrier systems and chimeric peptide technology) appear to be the most promising ones. This review combines a comprehensive overview on the current understanding of the transport mechanisms at the BBB with promising selected strategies published so far that can be applied to facilitate enhanced P/P drug delivery over the BBB. ß

Methods for chlorophenols (CPs) determination (with low limits of detection) that can be applied to real environmental samples (waters, sediments, soils, biological tissues) and food are reviewed. Special emphasis is given to sampling, storage conditions and the application of preconcentration techniques for the determination of CPs using chromatographic methods. Solid phase extraction, solid phase microextraction, stir bar sorptive extraction, liquid phase microextraction, dispersive liquid-liquid microextraction, liquid-liquid-liquid microextraction and purge and trap methods are considered. Methods for microwave and ultrasonic extraction of CPs from solid matrices are also focused.

Orchids are among the most diverse of the flowering plant families, with over 800 described genera and 25,000 species. Orchids are prized for their beautiful long lasting flowers exhibiting an incredible range of diversity in size, shape and colour. Today growing orchids is more than just a hobby, it is an international business covering around 8% of the world floriculture trade and has the potential to alter the economic landscape of a country. Large-scale multiplication of exquisite and rare hybrids using tissue culture techniques has helped orchids occupy a position as one of the top ten cut flowers. As orchids are outbreeders, their propagation using seeds leads to the production of heterozygous plants. Hence, protocols providing regeneration from various vegetative parts of the plants are needed. Though orchid micropropagation has shown spectacular development in the recent years, the wide spread use of micropropagation is believed to be still limited due to problems like exudation of phenolics from explants, transplantation to field, somaclonal variation etc. We endeavour to include the major investigations on explant-based orchid tissue culture starting from the pioneering works of Rotor [Rotor, G., 1949. A method of vegetative propagation of Phalaenopsis species and hybrids. Am. Orchid Soc. Bull. 18, 738-739] followed by Morel [Morel, G., 1960. Producing virus-free cymbidiums. Am. Orchid Soc. Bull. 29, 495-497] and Wimber [Wimber, D.E., 1963. Clonal multiplication of cymbidiums through tissue culture of the shoot meristem. Am. Orchid Soc. Bull. 32, 105-107] to date.

Olive mill wastewaters (OMW) are a potential source of biophenols, but they have a complex composition with many unknown phenolics. The analysis of purified methanol extracts from two Portuguese OMW by electrospray mass spectrometry in the negative mode showed [MÀH] À ions at m/z 539 and m/z 523, corresponding respectively to oleuropein and ligstroside isomers which contain the glucose unit linked to its aromatic moiety. Also, the fragmentation pathway of the [MÀH] À ions at m/z 863, 685 and 847 indicated the presence of a diglucoside derivative of the oleuropein isomer and of mono-and diglucosides of the ligstroside isomer, respectively. Moreover, the two OMW samples contained an elenoic derivative of the ion at m/z 685 and a degradation product (m/z 453) of the [MÀH] À ion at m/z 523. Future studies focusing on the abundance of these compounds on OMW, as well as their bioactivities, will determine their possible industrial exploitation.

The purpose of this work was to study the appropriateness of polyethylene bags for the preservation of explosive specimens. To this end, specimens of two types of dynamites, Goma-2 EC, containing nitroglycol (EGDN) and dinitrotoluene (DNT), and Goma-2 ECO, containing only EGDN, were placed individually inside bags and introduced into hermetically sealed glass jars, which were stored for a period of time. Losses of volatile compounds were studied by headspace analysis using Gas Chromatography coupled to Mass Spectrometry (GC-MS). The cross-contamination between dynamites was studied by using High Performance Liquid Chromatography with Mass Spectrometry (HPLC-MS) to analyze the extracts obtained after a sequential solvent extraction of these specimens. Polyethylene bags permit the loss of volatile compounds since EGDN and DNT were detected in the headspaces of the jars. Moreover, cross-contamination between dynamites was also demonstrated since DNT content decreased in the dynamite containing this compound and increased in the dynamite that had not contained it.

Stress, a common if unpredictable life event, can have pronounced effects on physiology and behavior. Individuals show wide variation in stress susceptibility and resilience, which are only partially explained by variations in coding genes. Developmental programing of the hypothalamic-pituitary-adrenal stress axis provides part of the explanation for this variance. Epigenetic approaches have successfully helped fill the explanatory gaps between the influences of gene and environment on stress responsiveness, and differences in the sequelae of stress across individuals and generations. Stress and the stress axis interacts bi-directionally with epigenetic marks within the brain. It is now clear that exposure to stress, particularly in early life, has both acute and lasting effects on these marks. They in turn influence cognitive function and behavior, as well as the risk for suicide and psychiatric disorders across the lifespan and, in some cases, unto future generations.

Adverse events Antipsychotic drug Autonomic Cardiovascular Side-effects Cardiovascular disease is the leading cause of death in people with severe mental disorders, and rates are proportionally greater than for other diseases such as cancer. Reports of sudden death in patients receiving antipsychotic treatment have raised concerns about the safety of antipsychotic drugs, leading to a number of recent changes in how such drugs are advertised and marketed. The majority of second generation antipsychotic drugs also have significant metabolic side-effects, such as weight gain, insulin resistance and hyperlipidemia, which may contribute indirectly to cardiovascular complications. As the use of antipsychotic drugs continues to expand into new indications and populations such as children and adolescents, a better understanding is needed of how antipsychotic drugs affect the cardiovascular system. Antipsychotic drugs interact with numerous receptors both centrally and peripherally, including monoamine receptors. The direct, non-specific pharmacological actions of antipsychotic drugs can lead to adverse cardiovascular effects, including orthostatic hypotension, tachycardia and ventricular arrhythmias. The mechanisms responsible for these antipsychotic-induced cardiovascular abnormalities have not been fully elucidated, but likely involve blockade of adrenergic or cholinergic receptors and hERG channels, in addition to impaired autonomic function. The direct and indirect effects of antipsychotic drugs on the cardiovascular system and their possible mechanisms of action are discussed in this review, where both preclinical and clinical findings are integrated.

Due to the limited numbers of PBMCs that can be obtained from the blood of individual mice, the key question whether central disease parameters such as onset, progression and severity correlate with the magnitude and cytokine quality of the T cell response in experimental autoimmune encephalomyelitis (EAE) has remained unanswered. Here we introduce an ELISPOT-based PBMC test system in which as little as 150 μl of murine blood are sufficient, allowing to bleed mice repeatedly while continuing to observe the clinical course of EAE. Using this technique, we demonstrate that longitudinal measurements of antigen-specific IFN-γ and IL-17 production in the blood are a highly suitable approach to predict the disease outcome in remitting-relapsing PLP:139-151-and chronic MOG:35-55-induced EAE of SJL/J and C57BL/6 mice, respectively. Our data propound cytokine monitoring as promising tool in the quest for more efficient diagnostic and prognostic options in human multiple sclerosis and other autoimmune diseases.

Endocannabinoids are endogenous agonists of cannabinoid receptors, and comprise amides, esters and ethers of long chain polyunsaturated fatty acids. Anandamide (N-arachidonoylethanolamine) and 2-arachidonoylglycerol are the best-studied members of this class of lipid mediators, and it is now widely accepted that their in vivo concentration and biological activity are largely dependent on a ''metabolic control.'' Therefore, the proteins that synthesize, transport and degrade endocannabinoids, and that together with the target receptors form the so-called ''endocannabinoid system,'' are the focus of intense research. This new system will be presented in this review, in order to put in a better perspective the impact of its modulation on Huntington's disease. In particular, the effect of agonists/antagonists of endocannabinoid receptors, or of inhibitors of endocannabinoid metabolism, will be discussed in the context of onset and progression of Huntington's disease, and will be compared with other neurodegenerative diseases like Parkinson's disease, Alzheimer's disease, and amyotropic lateral sclerosis. Also the plastic changes of endocannabinoids in multiple sclerosis will be reviewed, as a paradigm of their impact in neuroinflammatory disorders. #

Modern trends in analytical chemistry are towards the simplification and miniaturization of sample preparation, as well as the minimization of organic solvent used. In view of this aspect, several novel micro-extraction techniques are being developed in order to reduce the analysis step, increase the sample throughput and to improve the quality and the sensitivity of analytical methods. One of the emerging techniques in this area is liquid-phase micro-extraction (LPME). It is a miniaturized implementation of conventional liquid/liquid extraction (LLE) in which only microliters of solvents are used instead of several hundred milliliters in LLE. It is quick, inexpensive and can be automated. In the last few years, LPME has been combined with liquid chromatography (LC) and capillary electrophoresis (CE), besides the generally used coupling to gas chromatography (GC), and has been applied to various matrices, including biological, environmental, and food samples. This work is aimed at providing an overview of the major developments of LPME, coupled with chromatography and CE, as reported in the literature. The paper will focus on the application of the technique to different matrices and the aim is to reveal the panorama of opportunities and to try to indicate the potential of LPME in pesticide analysis. A critical review of the first applications to pesticide analyses is presented in the main part of the manuscript. The optimization of LPME as well as advantages and disadvantages are discussed. It is concluded that, because of its high preconcentration factor, LPME can be introduced with benefit into water analysis for several pesticide groups. In particular, the application of LPME to non-polar pesticides in environmental analysis appears to be promising. However, similar to other micro-extraction techniques, such as solid phase micro-extraction (SPME), serious limitations still remain when analyzing semi-solid and solid environmental, food or biological matrices and/or highly polar compounds. Thus, other pre-concentration techniques may be a good alternative if an analytical problem cannot be sufficiently dealt with LPME.