Matrix metalloproteinases in the adult brain physiology: a link between c-Fos,AP-1 and remodeling of neuronal connections?

Matrix metalloproteinases (MMPs), together with their endogenous inhibitors (TIMPs) form an enzymatic system that plays an important role in a variety of physiological and pathological conditions. These proteins are also expressed in the brain, especially under pathological conditions, in which glia as well as invading inflammatory cells provide the major source of the MMP activity. Surprisingly little is known about the MMP function(s) in adult neuronal physiology. This review describes available data on this topic, which is presented in a context of knowledge about the MMP/TIMP system in other organs as well as in brain disorders. An analysis of the MMP and TIMP expression patterns in the brain, along with a consideration of their regulatory mechanisms and substrates, leads to the proposal of possible roles of the MMP system in the brain. This analysis suggests that MMPs may play an important role in the neuronal physiology, especially in neuronal plasticity, including their direct participation in the remodeling of synaptic connections-a mechanism pivotal for learning and memory.     

The effect of EDTA and EDDS on lead uptake and localization in hydroponically grown Pisum sativum L.

Pisum sativum plants were treated for 3 days with an aqueous solution of 100 μM Pb(NO₃)₂ or with a mixture of lead nitrate and ethylenediaminetetraacetic acid (EDTA) or [S,S]-ethylenediaminedisuccinic acid (EDDS) at equimolar concentrations. Lead decline from the incubation media and its accumulation and localization at the morphological and ultrastructural levels as well as plant growth parameters (root growth, root and shoot dry weight) were estimated after 1 and 3 days of treatment. The tested chelators, especially EDTA, significantly diminished Pb uptake by plants as compared to the lead nitrate-treated material. Simultaneously, EDTA significantly enhanced Pb translocation from roots to shoots. In the presence of both chelates, plant growth parameters remained considerably higher than in the case of uncomplexed Pb. Considerable differences between the tested chelators were visible in Pb localization both at the morphological and ultrastructural level. In Pb+EDTA-treated roots, lead was mainly located in the apical parts, while in Pb+EDDS-exposed material Pb was evenly distributed along the whole root length. Transmission electron microscopy and EDS analysis revealed that in meristematic cells of the roots incubated in Pb+EDTA, large electron-dense lead deposits were located in vacuoles and small granules were rarely noticed in cell walls or cytoplasm, while after Pb+EDDS treatment metal deposits were restricted to the border between plasmalemma and cell wall. Such results imply different ways of transport of those complexed Pb forms.     

HPLC columns partition by chemometric methods based on peptides retention

In recent years, multivariate techniques have been utilized to evaluate reversed-phase high-performance liquid chromatographic data. In the present study, 11 high-performance liquid chromatography (HPLC) columns were divided into several groups according to the retention factors of 12 peptides. Principal component analysis (PCA) and cluster analysis (CA) were used in column and peptides' comparison and grouping. CA results indicated that all stationary phases may be generally grouped into several clusters, due to stationary phase structure and properties. On the other hand, interesting results were obtained with the use of PC. There is almost linear relationship between classified HPLC columns in the space of new PCs, which is connected with meaning of the PC's reflected in their loading values. The first component describes non-polar properties of peptides, whereas the second component is loaded with polar peptides having much lower logP values. PCA and CA were also used in peptides comparison however, complete explanation of peptides grouping still remains unclear.     

Aftiphilin is a component of the clathrin machinery in neurons

Aftiphilin was identified through a database search for proteins containing binding motifs for the gamma-ear domain of clathrin adaptor protein 1 (AP-1). Here, we demonstrate that aftiphilin is expressed predominantly in brain where it is enriched on clathrin-coated vesicles. In addition to eight gamma-ear-binding motifs, aftiphilin contains two WXXF-acidic motifs that mediate binding to the alpha-ear of clathrin adaptor protein 2 (AP-2) and three FXXFXXF/L motifs that mediate binding to the alpha- and beta2-ear. We demonstrate that aftiphilin uses these motifs for interactions with AP-1 and AP-2 and that it immunoprecipitates these APs but not AP-3 or AP-4 from brain extracts. Aftiphilin demonstrates a brefeldin A sensitive localization to the trans-Golgi network in hippocampal neurons where it co-localizes with AP-1. Aftiphilin is also found at synapses where it co-localizes with synaptophysin and AP-2. Our data suggest a role for aftiphilin in clathrin-mediated trafficking in neurons.     

RS-alpha-lipoic acid protects cholinergic cells against sodium nitroprusside and amyloid-beta neurotoxicity through restoration of acetyl-CoA level

The work presented here verifies the hypothesis that RS-alpha-lipoic acid may exert its cholinoprotective and cholinotrophic activities through the maintenance of appropriate levels of acetyl-CoA in mitochondrial and cytoplasmic compartments of cholinergic neurons. Sodium nitroprusside (SNP) and amyloid-beta decreased pyruvate dehydrogenase, choline acetyltransferase activities, acetyl-CoA content in mitochondria and cytoplasm, as well as increased fraction of non-viable, trypan blue positive cells in cultured differentiated cholinergic SN56 neuroblastoma cells. Lipoic acid totally reversed toxin-evoked suppression of choline acetyltrasferase and pyruvate dehydrogenase activities, as well as mitochondrial and cytoplasmic acetyl-CoA levels, and partially attenuated increase of cell mortality. Significant negative correlations were found between enzyme activities, acetyl-CoA levels and cell mortality in different neurotoxic and neuroprotective conditions employed here. The level of cytoplamic acetyl-CoA correlated with mitochondrial acetyl-CoA, whereas choline acetyltransferase activity followed shifts in cytoplasmic acetyl-CoA. Thus, we conclude that, in cholinergic neurons, particular elements of the pyruvate-acetyl-CoA-acetylcholine pathway form a functional unit responding uniformly to nerotoxic and neuroprotectory conditions.     

Evidence for significant heritability of apoptotic and cell cycle responses to ionising radiation

Genetic factors are likely to affect individual cancer risk, but few quantitative estimates of heritability are available. Public health radiation protection policies do not in general take this potentially important source of variation in risk into account. Two surrogate cellular assays that relate to cancer susceptibility have been developed to gain an insight into the role of genetics in determining individual variation in radiosensitivity. These flow cytometric assays for apoptosis induction and cell cycle delay following radiation are sufficiently sensitive to distinguish lymphocytes from a healthy donor population from those of a sample of obligate carriers of ATM mutations (P = 0.01 and P = 0.02, respectively). Analysis of 54 unselected twin pairs (38 dizygotic, 16 monozygotic) indicated much greater intrapair correlation in response in monozygotic than in dizygotic pairs. Structural equation modelling indicated that models including unique environmental factors only fitted the data less well than those incorporating two or more of additive genetic factors, common environmental factors and unique environmental factors. A model incorporating additive genetic factors and unique environmental factors yielded estimates of heritability for the two traits of 68% (95% CI 40–82%, cell cycle) and 59% (95% CI 22–79%, apoptosis). Thus, these data suggest that genetic factors contribute significantly to human variation in these two measures of radiosensitivity that relate to cancer susceptibility.     

Small things are important: the value of singular point elements for birds in agricultural landscapes

Farmland birds belong to the most endangered group of vertebrates in Europe. They are an important component of farmland biodiversity considering the numerous functions they perform (e.g. seed dispersal, improving germination, increasing gene flow, nutrient recycling, and pest control). Therefore, their decline imposes substantial risks on agricultural ecosystems. In general, farmland bird conservation includes land-use and management alterations leading to less-intensive farming and land-sparing for breeding habitats (e.g. agri-environment-climate schemes, and organic farming). However, theoretical concepts describing farmland biodiversity maintenance and applied conservation measures usually ignore the role of singular, often very small, natural or man-made elements in an agricultural landscape. These elements play a role in the populations of certain species, their biology and in the general species richness of farmland. Furthermore, the importance of these elements has never been empirically tested, which means that conservationists and practitioners are not aware of their measurable value for birds. Herein, we define and identify singular point elements in the agricultural landscape (SPELs) which are potentially important for breeding farmland birds. We also describe each SPEL and evaluate its importance for birds in farmland based on a systematic review of the available literature. Using a horizon-scanning technique, we then polled field ornithologists about their personal observations of birds in relation to SPELs and the evaluation of the potential roles of such structures for birds. We identified 17 SPELs that vary in naturalness and age: singular trees, singular shrubs, erratic boulders, puddles, electricity pylons, wind turbines, spiritual sites, hunting platforms, fence and border posts, wells, road signs, scarecrows, piles of manure, piles of brushwood/branches, piles of stones/debris, piles of lime, and haystacks. Analysis of the literature revealed knowledge gaps, because some SPELs are frequently mentioned in ecological studies (e.g. trees, shrubs, pylons), but others such as spiritual sites, stones, hunting platforms, wells, road signs, or piles of lime are ignored. Despite the fact that some authors incorporate the effects of some SPELs in their studies, little research to date has aimed to assess the impact of various SPELs on farmland bird species numbers and distribution. Horizon scanning revealed that ornithologists often observe birds on various SPELs and thus, attribute to SPELs many functions that are important for maintaining bird populations. Horizon scanning also highlighted the importance of SPELs for many declining bird species and suggested possible mitigation of negative changes in the agricultural landscape by retaining SPELs within fields. We suggest that a better understanding of the role of SPELs for farmland birds is required. We also recommend that SPELs are considered as a potential tool for the conservation of birds, and existing conservation programs such as agri-environment-climate schemes and organic farming should be updated accordingly. Finally, we suggest that SPELs are included in predictive models that evaluate habitat suitability for farmland biodiversity.     

Successful elimination of non-neural cells and unachievable elimination of glial cells by means of commonly used cell culture manipulations during differentiation of GFAP and SOX2 positive neural progenitors (NHA) to neuronal cells

Background  

Although extensive research has been performed to control differentiation of neural stem cells – still, the response of those cells to diverse cell culture conditions often appears to be random and difficult to predict. To this end, we strived to obtain stabilized protocol of NHA cells differentiation – allowing for an increase in percentage yield of neuronal cells.