Characteristics of brain gastrin/cholecystokinin-binding proteins

Gastrin/cholecystokinin-binding proteins were purified using the column affinity chromatography on immobilized pig tetragastrin and cholecystokinin. Immunoblotting analysis of different human tissue extracts with specific antisera obtained against gastrin-binding proteins was performed. It was found that high molecular weight polypeptide zones of 120 kDa and 35 kDa were characteristic of the brain only. Autoantisera of patients with type A gastric disease reacted with some gastrin/cholecystokinin-binding proteins in human brain and mucosa including human brain polypeptide of 120 kDa. It is supposed that there are neurospecific gastrin-binding proteins (possibly gastrin/cholecystokinin receptors in the brain).     

Crosses of two independently derived transgenic mice demonstrate functional complementation of the genes encoding heavy (HLA-B27) and light (beta 2-microglobulin) chains of HLA class I antigens

In man a number of diseases are associated with certain alleles of MHC antigens. The most pronounced example is ankylosing spondylitis, which is strongly associated with HLA-B27. As a first step towards a model system to study the basis of this association, transgenic mice were generated that showed cell surface expression of the HLA-B27 antigen biochemically indistinguishable from HLA-B27 antigen expressed on human cells. This result was obtained by crossing two independently derived strains of mice, one of which is transgenic for the HLA-B27 heavy chain gene, and the other carrying and expressing the human beta 2m gene. Examination of HLA-B27 and human beta 2m mRNA in various tissues shows the two genes to be expressed in a coordinate fashion. The mRNA levels follow those of endogenous H-2 Class I genes.     

Use of drug-coated stents to treat coronary bifurcation lesions

Bifurcation stenoses are one of the most complex categories for endovascular treatment as such lesion requires that the lumen of both a great vessel and a lateral branch be recovered. Introduction of drug-coated stents into clinical practice has changed surgical approaches in this type of an atherosclerotic lesion. The use of two drug-coated endoprostheses or more within the afflicted segment to stent a great vessel or lateral branch does not increase a risk for restenosis. The considerable opportunities of endovascular technologies and the absence of a unified surgical approach in this type of lesion generate a need for systematizing the experience accumulated. The purpose of the present study is to develop an algorithm of bifurcation stenting in different types of X-ray anatomy and morphology of a coronary lesion.     

Photosystem II associated carbonic anhydrase activity in higher plants is situated in core complex

The thylakoid membrane containing photosystem II (PSII membranes) from pea and wheat leaves catalyzed the reaction of CO2 hydration with low rate, which increased after their incubation either with Triton X-100, up to Triton/chlorophyll ratio 1:1, or 1 M CaCl2. The presence of the inhibitor of CAs, p-aminomethylbenzensulfonamide (mafenide), at the start line in the course of electrophoresis of PSII membranes solubilized by n-dodecyl-beta-maltoside (DM) decreased the amount of PSII core complex in the gel. The elution of PSII core complex from the column with immobilized mafenide occurred only either by mafenide or another inhibitor of CAs, ethoxyzolamide. The above results led to a conclusion that membrane-bound CA activity associated with PSII is situated in the core complex.     

Modulation of the homophilic interaction between the first and second Ig modules of neural cell adhesion molecule by heparin

The second Ig module (IgII) of the neural cell adhesion molecule (NCAM) is known to bind to the first Ig module (IgI) of NCAM (so-called homophilic binding) and to interact with heparan sulfate and chondroitin sulfate glycoconjugates. We here show by NMR that the heparin and chondroitin sulfate-binding sites (HBS and CBS, respectively) in IgII coincide, and that this site overlaps with the homophilic binding site. Using NMR and surface plasmon resonance (SPR) analyses we demonstrate that interaction between IgII and heparin indeed interferes with the homophilic interaction between IgI and IgII. Accordingly, we show that treatment of cerebellar granule neurons (CGNs) with heparin inhibits NCAM-mediated outgrowth. In contrast, treatment with heparinase III or chondroitinase ABC abrogates NCAM-mediated neurite outgrowth in CGNs emphasizing the importance of the presence of heparan/chondroitin sulfates for proper NCAM function. Finally, a peptide encompassing HBS in IgII, termed the heparin-binding peptide (HBP), is shown to promote neurite outgrowth in CGNs. These observations indicate that neuronal differentiation induced by homophilic NCAM interaction is modulated by interactions with heparan/chondroitin sulfates.     

Blocking variant surface glycoprotein synthesis alters endoplasmic reticulum exit sites/Golgi homeostasis in Trypanosoma brucei

The predominant secretory cargo of bloodstream form Trypanosoma brucei is variant surface glycoprotein (VSG), comprising ~10% total protein and forming a dense protective layer. Blocking VSG translation using Morpholino oligonucleotides triggered a precise pre‐cytokinesis arrest. We investigated the effect of blocking VSG synthesis on the secretory pathway. The number of Golgi decreased, particularly in post‐mitotic cells, from 3.5 ± 0.6 to 2.0 ± 0.04 per cell. Similarly, the number of endoplasmic reticulum exit sites (ERES) in post‐mitotic cells dropped from 3.9 ± 0.6 to 2.7 ± 0.1 eight hours after blocking VSG synthesis. The secretory pathway was still functional in these stalled cells, as monitored using Cathepsin L. Rates of phospholipid and glycosylphosphatidylinositol‐anchor biosynthesis remained relatively unaffected, except for the level of sphingomyelin which increased. However, both endoplasmic reticulum and Golgi morphology became distorted, with the Golgi cisternae becoming significantly dilated, particularly at the trans‐face. Membrane accumulation in these structures is possibly caused by reduced budding of nascent vesicles due to the drastic reduction in the total amount of secretory cargo, that is, VSG. These data argue that the total flux of secretory cargo impacts upon the biogenesis and maintenance of secretory structures and organelles in T. brucei, including the ERES and Golgi.      

Mid- and late-Holocene vegetation history,climate and human impact in the forest-steppe ecotone of European Russia: new data and a regional synthesis

Appropriate management of contemporary environments requires knowledge of their long-term history. We use palaeoecological data to explore how contemporary forest-steppe environments have been shaped by climate change and human impacts through the Holocene using the western Mid-Russian Upland as a case-study. Our paper presents new reconstructions of Mid- and Late Holocene climate, vegetation dynamics and local environmental change based on pollen, plant macrofossil and testate amoeba records from a site at Selikhovo (Mid-Russian Upland, Russia). Eutrophic fen vegetation dominated by Phragmites australis developed around 6800 cal year BP and has been resilient to episodes of local burning and variable input of mineral material through the Holocene. New and previously-published data show that the boundary between broadleaf forest and steppe occupied a similar position to present during the period 7000–4800 cal year BP, despite a warmer and drier climate, but shifted to the south following climate cooling and an increase in precipitation from 4800–2500 cal year BP. A subsequent decline in woodland cover was caused by both climate change and human impacts, with human activity becoming increasingly significant over the last two millennia. Prior to major human disturbance (about 1700 cal year BP) the landscape was dominated by mixed broadleaf-pine forests with some spruce covering about 60 % of the study area. Our results emphasize the variability of steppe-forest habitats over long time periods and the need to consider human impacts and climate change when setting targets for habitat conservation.