Monocyte NOTCH2 expression predicts interferon-beta immunogenicity in multiple sclerosis patients

Multiple sclerosis (MS) is an autoimmune disease characterized by CNS inflammation leading to demyelination and axonal damage. IFN-β is an established treatment for MS; however, up to 30% of IFN-β–treated MS patients develop neutralizing antidrug antibodies (nADA), leading to reduced drug bioactivity and efficacy. Mechanisms driving antidrug immunogenicity remain uncertain, and reliable biomarkers to predict immunogenicity development are lacking. Using high-throughput flow cytometry, NOTCH2 expression on CD14+ monocytes and increased frequency of proinflammatory monocyte subsets were identified as baseline predictors of nADA development in MS patients treated with IFN-β. The association of this monocyte profile with nADA development was validated in 2 independent cross-sectional MS patient cohorts and a prospective cohort followed before and after IFN-β administration. Reduced monocyte NOTCH2 expression in nADA+ MS patients was associated with NOTCH2 activation measured by increased expression of Notch-responsive genes, polarization of monocytes toward a nonclassical phenotype, and increased proinflammatory IL-6 production. NOTCH2 activation was T cell dependent and was only triggered in the presence of serum from nADA+ patients. Thus, nADA development was driven by a proinflammatory environment that triggered activation of the NOTCH2 signaling pathway prior to first IFN-β administration

Species Specificity in Major Urinary Proteins by Parallel Evolution

Species-specific chemosignals, pheromones, regulate social behaviors such as aggression, mating, pup-suckling, territory establishment, and dominance. The identity of these cues remains mostly undetermined and few mammalian pheromones have been identified. Genetically-encoded pheromones are expected to exhibit several different mechanisms for coding 1) diversity, to enable the signaling of multiple behaviors, 2) dynamic regulation, to indicate age and dominance, and 3) species-specificity. Recently, the major urinary proteins (Mups) have been shown to function themselves as genetically-encoded pheromones to regulate species-specific behavior. Mups are multiple highly related proteins expressed in combinatorial patterns that differ between individuals, gender, and age; which are sufficient to fulfill the first two criteria. We have now characterized and fully annotated the mouse Mup gene content in detail. This has enabled us to further analyze the extent of Mup coding diversity and determine their potential to encode species-specific cues

A qualitative investigation of major urinary proteins in relation to the onset of aggressive behavior and dispersive motivation in male wild house mice (Mus musculus domesticus)

The physiological basis for population differentiation of dispersal timing during individual development in male wild house mice is still unknown. As major urinary proteins (MUPs) are known to convey information about competitive ability in male mice, we examined individual MUP profiles defined by isoelectric-focusing (IEF) patterns in relation to developmental timing of dispersive motivation. As an experimental paradigm marking the development of the dispersal propensity, we used agonistic onset between litter mate brothers when kept in pairs under laboratory conditions. Agonistic onset is known to reflect the initiation of dispersive motivation. Hence, we compared individual MUP IEF patterns between fraternal pairs that did or did not develop agonistic relationships before the age of 2 months. Urine was collected on the day of weaning and at the beginning of adulthood. We investigated whether there was a significant co-occurrence of particular MUP IEF patterns with the agonistic onset in male mice. We assumed that, based on this co-occurrence, particular MUP IEF patterns and/or a particular dynamic of MUP IEF expression from weaning to adulthood may be considered a physiological predictor of a specific behavioral strategy in male mice (i.e. submissive-philopatric or agonistic-dispersive strategy). We found that agonistic males expressed more MUP IEF bands than amicable ones at weaning, but these differences disappeared later on. The presence of two particular IEF bands at weaning was significantly associated with early agonistic onset. Our study suggests that MUPs could have a predictive value for the onset of aggressive behavior and dispersal tendency in male wild house mice

Crystallization and preliminary X-ray analysis of porcine muscle prolyl oligopeptidase

Prolyl oligopeptidase from pig muscle has been crystallized in complex with an inhibitor, using PEG 8000 and calcium acetate as precipitants. The crystals are orthorombic and the space group is P212121 with cell dimensions a = 111.8, b = 101.8, c = 72.4 Å. The asymmetric unit contains a single chain of prolyl oligopeptidase, corresponding to a specific volume of 2.55 Å3 Da-1 and a solvent content of 52%. The observed diffraction pattern extends to 2.3 Å resolution and the native crystals are well suited for structural analysis by X-ray diffraction methods. © 1998 International Union of Crystallography Printed in Great Britain - all rights reserved

Role of electrostatics at the catalytic metal binding site in xylose isomerase action: Ca2+-inhibition and metal competence in the double mutant D254E/D256E

The catalytic metal binding site of xylose isomerase from Arthrobacter B3728 was modified by protein engineering to diminish the inhibitory effect of Ca2+ and to study the competence of metals on catalysis. To exclude Ca2+ from Site 2 a double mutant D254E/D256E was designed with reduced space available for binding. In order to elucidate structural consequences of the mutation the binary complex of the mutant with Mg2+ as well as ternary complexes with bivalent metal ions and the open-chain inhibitor xylitol were crystallized for x-ray studies. We determined the crystal structures of the ternary complexes containing Mg2+, Mn2+, and Ca2+ at 2.2 to 2.5 \uc5 resolutions, and refined them to R factors of 16.3, 16.6, and 19.1, respectively. We found that all metals are liganded by both engineered glutamates as well as by atoms O1 and O2 of the inhibitor. The similarity of the coordination of Ca2+ to that of the cofactors as well as results with Be2+ weaken the assumption that geometry differences should account for the catalytic noncompetence of this ion. Kinetic results of the D254E/D256E mutant enzyme showed that the significant decrease in Ca2+ inhibition was accompanied by a similar reduction in the enzymatic activity. Qualitative argumentation, based on the protein electrostatic potential, indicates that the proximity of the negative side chains to the substrate significantly reduces the electrostatic stabilization of the transition state. Furthermore, due to the smaller size of the catalytic metal site, no water molecule, coordinating the metal, could be observed in ternary complexes of the double mutant. Consequently, the proton shuttle step in the overall mechanism should differ from that in the wild type. These effects can account for the observed decrease in catalytic efficiency of the D254E/D256E mutant enzyme

РЕФЕРЕНТНЫЕ ЗНАЧЕНИЯ СКОРОСТИ РАСПРОСТРАНЕНИЯ ПУЛЬСОВОЙ ВОЛНЫ ПО АОРТЕ У ЗДОРОВЫХ ДЕТЕЙ В ВОЗРАСТЕ ОТ 3 ДО 18 ЛЕТ

Objective: The measurement of aortic pulse wave velocity (PWVao) is an accepted marker in stratifying individual cardiovascular risk in adults. There is an increasing volume of evidence concerning impaired vascular function in different diseases in paediatric populations, but, unfortunately, only a few studies are available on the measurement of normal PWVao values in children. The aim of our study was to determine the reference values of PWVao in a large healthy population using a newly developed technique. Methods: Three thousand, three hundred and seventyfour healthy individuals (1802 boys) aged 3–18 years were examined by an invasively validated, occlusive, oscillometric device. Results: The mean PWVao values increased from 5.5_0.3 to 6.5_0.3 m/s (P<0.05) in boys and from 5.6_0.3 to 6.4_0.3 m/s (P<0.05) in girls. The increase, however, was not constant, and the values exhibited a flat period between the ages of 3 and 8 years in both sexes. The first pronounced increase occurred at the age of 12.1 years in boys and 10.4 years in girls. Moreover, between the ages of 3 and 8 years, the brachial SBP and mean blood pressures increased continuously and gradually, whereas the PWVao remained unchanged. By contrast, beyond the age of 9 years, blood pressure and aortic stiffness trends basically moved together. Conclusion: Our study provides the largest database to date concerning arterial stiffness in healthy children and adolescents between the ages of 3 and 18 years, and the technology adopted proved easy to use in large paediatric populations, even at a very young age.  Цель. Измерение скорости распространения пульсовой волны (СРПВ) по аорте является маркером стратификации риска развития сердечно-сосудистых заболеваний у взрослых. Доказано, что у детей с различными заболеваниями отмечается ухудшение сосудистой функции, но, к сожалению, только в нескольких исследованиях определены нормальные значения скорости распространения пульсовой волны у детей. Цель нашего исследования ― определить референтные значения СРПВ у здоровых детей,  используя новейшие технологии. Пациенты и методы. С помощью неинвазивного  окклюзионного осциллометрического аппарата обследовано 3374 здоровых ребенка (из них 1802 мальчики) в возрасте от 3 до 18 лет. Результаты. Средние значения СРПВ увеличиваются от 5,5±0,3 до 6,5±03 м/с (р<0,05) у мальчиков и с 5,6±0,3 до 6,4±03 м/с (р<0,05) у девочек. Тем не менее данное повышение не постоянное. У детей обоих полов в возрасте от 3 до 8 лет увеличения СРПВ не отмечается. Первое отчетливое повышение СРПВ происходит в возрасте 12,1 лет у мальчиков и в 10,4 года у девочек.  В возрасте от 3 до 8 лет отмечалось непрерывное постепенное нарастание систолического артериального давления, измеренного на плече, и среднего артериального давления, при этом показатели СРПВ оставались неизменными. С 9 лет показатели артериального давления и ригидности аорты изменялись одновременно. Заключение. В проведенном исследовании представлена самая большая на сегодняшний день база данных по артериальной ригидности, полученная у здоровых детей и подростков в возрасте от 3 до 18 лет с помощью простого исследования, адаптированного для широкого применения в педиатрической практике, даже у детей раннего возраста