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41.
42.
A plant lectin was isolated from barley (Hordeum vulgare) coleoptiles using acidic extraction and different chromatographic methods. Sequencing of more than 50% of the protein sequence
by Edman degradation confirmed a full-length cDNA clone. The subsequently identified open reading frame encodes for a 15 kDa
protein which could be found in the soluble fraction of barley coleoptiles. This protein exhibited specificity towards mannose
sugar and is therefore, accordingly named as Horcolin (Hordeum vulgare coleoptile lectin). Database searches performed with
the Horcolin protein sequence revealed a sequence and structure homology to the lectin family of jacalin-related lectins.
Together with its affinity towards mannose, Horcolin is now identified as a new member of the mannose specific subgroup of
jacalin-related lectins in monocot species. Horcolin shares a high amino acid homology to the highly light-inducible protein
HL#2 and, in addition to two methyl jasmonic acid-inducible proteins of 32.6 and 32.7 kDa where the jasmonic acid-inducible
proteins are examples of bitopic chimerolectins containing a dirigent and jacalin-related domain. Immunoblot analysis with
a cross-reactive anti-HL#2 antibody in combination with Northern blot analysis of the Horcolin cDNA revealed tissue specific
expression of Horcolin in the coleoptiles. The function of Horcolin is discussed in the context of its particular expression
in coleoptiles and is then compared to other lectins, which apparently share a related response to biotic or abiotic stress
factors. 相似文献
43.
Jarutat T Nickels C Frisch C Stellmacher F Hofig KP Knappik A Merz H 《Biological chemistry》2007,388(6):651-658
We describe the direct isolation of specific antibodies on formalin-fixed, paraffin-embedded (FFPE) tissue. The technique involves subtractive selection of a large and highly diverse combinatorial human antibody phage library (HuCAL) on lymphocyte FFPE tissue sections. Tissue sections from normal human tonsil tissue were used to deplete the library of binders to most housekeeping proteins. Mantle-cell lymphoma tissue was used for positive selection and enrichment of mantle cell or tumor-specific antibody phage. We established a high-throughput immunohistochemical method for screening of antibody clones selected from FFPE tissue. One recombinant antibody showed specific staining for interfollicular and mantle cells in FFPE tissue. Immunoprecipitation with this antibody and subsequent mass spectrometric analysis revealed specificity for vimentin. 相似文献
44.
Achim Brockmeier Manuel Skopnik Brigitte Koch Christian Herrmann Stefan Welti Klaus Scheffzek 《Biochemical and biophysical research communications》2009,388(4):630-636
Eubacteria can import and simultaneously phosphorylate a range of different carbohydrates by means of sugar specific phosphoenolpyruvate (PEP) dependent sugar phosphotransferase systems (PTSs). Here, we report the biochemical characterization of the gluconate specific PTS component EIIAgnt from Enterococcus faecalis and its unexpectedly strong complex with EIIBgnt. We analyze the activity of the complex regarding phosphoryl transfer using kinetic measurements and demonstrate by mutagenesis that His-9 of EIIAgnt is essential for this process and represents most likely the phosphoryl group carrier of EIIAgnt. With a combination of isothermal titration calorimetry (ITC), analytical ultracentrifugation (AUC), native gel electrophoresis and chemical crosslinking experiments we show that EIIAgnt and EIIBgnt form a strong 2:2 heterotetrameric complex, which seems to be destabilized upon phosphorylation of EIIBgnt. 相似文献
45.
Cristina L. Vázquez Thomas R. Lerner Bahram Kasmapour Gang Pei Achim Gronow Maria V. Bianco Federico C. Blanco Christopher K. E. Bleck Robert Geffers Fabiana Bigi Wolf‐Rainer Abraham Maximiliano G. Gutierrez 《Cellular microbiology》2014,16(9):1425-1440
Some intracellular bacteria are known to cause long‐term infections that last decades without compromising the viability of the host. Although of critical importance, the adaptations that intracellular bacteria undergo during this long process of residence in a host cell environment remain obscure. Here, we report a novel experimental approach to study the adaptations of mycobacteria imposed by a long‐term intracellular lifestyle. Selected Mycobacterium bovis BCG through continuous culture in macrophages underwent an adaptation process leading to impaired phenolic glycolipids (PGL) synthesis, improved usage of glucose as a carbon source and accumulation of neutral lipids. These changes correlated with increased survival of mycobacteria in macrophages and mice during re‐infection and also with the specific expression of stress‐ and survival‐related genes. Our findings identify bacterial traits implicated in the establishment of long‐term cellular infections and represent a tool for understanding the physiological states and the environment that bacteria face living in fluctuating intracellular environments. 相似文献
46.
Angela Schippers Silke Mateika Blair Prochnow Achim D. Gruber Werner Müller Ursula Frischmann 《Mammalian genome》2008,19(4):279-291
EUMORPHIA (European Union Mouse Research for Public Health and Industrial Application) is a research program involved in developing new approaches in phenotyping, mutagenesis, and informatics to improve
characterization of mouse models for understanding human physiology and disease. Secondary screen experiments include the
development of assays to identify mice with altered susceptibility or resistance to infections. In this context we developed
a new model and established a standard operating procedure for the experimental infection of mice with Yersinia (Y.) enterocolitica. In contrast with previous studies that dealt with high-pathogenic Y. enterocolitica, we used the low-pathogenic Y. enterocolitica strain E40 to analyze differences in the immune response of four strains of inbred mice (BALB/c, C3H/HeN, 129P2, C57BL/6)
after oral infection. The determination of colony-forming units in Peyer’s patches and histologic analysis supported the observations
that BALB/c are less able to ameliorate the infection within 21 days. The immune defense of C57BL/6 mice against Yersinia was the most effective resulting in a nearly complete elimination of bacteria after 21 days. C3H/HeN and 129P2 were intermediate.
Analysis of serum immunoglobulins (Ig) by Luminex showed a significant increase of IgG2b levels 21 days after infection in
all four inbred strains. The other immunoglobulins remained nearly constant. Our infection model discriminates between the
efficiency of an infection at an early time point (3 days) and immunity at a later time point (21 days). It is furthermore
an appropriate model to characterize genetic differences in resistance and immunity of inbred and mutant mouse lines. 相似文献
47.
As research progress has led to programs for the elimination of onchocerciasis as a public health problem, research must now be intensified to protect elimination efforts. A profound understanding of the immunology in the human-parasite relationship is required for predicting the impacts of an altered immune response in a population post-microfilaricide treatment, and for the development of a vaccine against onchocerciasis, a highly desirable tool to guarantee sustained elimination success. This article summarizes the recent advancements in understanding the human immune mechanisms against onchocerciasis, and focuses on the new concept of T-cell suppressor responses as a major counterbalance mechanism for effector responses driven by T helper 1 and T helper 2 cells against the filarial worms. 相似文献
48.
SUMMARY 1. We investigated the effect of temperature on chemical signalling in a predator–prey model system (planktivorous fish and Daphnia galeata ). Life-history changes in Daphnia in response to chemical cues (kairomones) derived from fish have become a paradigm for chemically induced anti-predator defences.
2. As temperature can affect both predator and prey, we carried out two experiments to disentangle these effects. In order to test for temperature effects on the predator, we kept prey at a single temperature and exposed them to kairomones from fish exposed to two different temperatures. Daphnia exhibited a higher intrinsic rate of population increase ( r ) when exposed to fish kairomones produced at high rather than low temperature. Assuming a positive correlation between r (because of an earlier maturation and/or increased clutch sizes) and kairomone concentration, our results suggest that kairomone production increases with rising temperature.
3. In the second experiment, to study the influence of temperature on the prey, Daphnia were kept at two different temperatures and exposed to fish kairomones produced at one constant temperature. We found no interaction between the effects of fish kairomone and temperature on Daphnia life history, suggesting that temperature does not directly alter life-history responses to fish kairomones.
4. Our results suggest that temperature influences Daphnia life history through its effects on fish kairomone concentration, but that temperature does not affect the strength of the response of Daphnia to the presence of fish. 相似文献
2. As temperature can affect both predator and prey, we carried out two experiments to disentangle these effects. In order to test for temperature effects on the predator, we kept prey at a single temperature and exposed them to kairomones from fish exposed to two different temperatures. Daphnia exhibited a higher intrinsic rate of population increase ( r ) when exposed to fish kairomones produced at high rather than low temperature. Assuming a positive correlation between r (because of an earlier maturation and/or increased clutch sizes) and kairomone concentration, our results suggest that kairomone production increases with rising temperature.
3. In the second experiment, to study the influence of temperature on the prey, Daphnia were kept at two different temperatures and exposed to fish kairomones produced at one constant temperature. We found no interaction between the effects of fish kairomone and temperature on Daphnia life history, suggesting that temperature does not directly alter life-history responses to fish kairomones.
4. Our results suggest that temperature influences Daphnia life history through its effects on fish kairomone concentration, but that temperature does not affect the strength of the response of Daphnia to the presence of fish. 相似文献
49.
50.
In mammals, the circadian pacemaker, which controls daily rhythms, is located in the suprachiasmatic nucleus (SCN). Circadian oscillations are generated in individual SCN neurons by a molecular regulatory network. Cells oscillate with periods ranging from 20 to 28 h, but at the tissue level, SCN neurons display significant synchrony, suggesting a robust intercellular coupling in which neurotransmitters are assumed to play a crucial role. We present a dynamical model for the coupling of a population of circadian oscillators in the SCN. The cellular oscillator, a three-variable model, describes the core negative feedback loop of the circadian clock. The coupling mechanism is incorporated through the global level of neurotransmitter concentration. Global coupling is efficient to synchronize a population of 10,000 cells. Synchronized cells can be entrained by a 24-h light-dark cycle. Simulations of the interaction between two populations representing two regions of the SCN show that the driven population can be phase-leading. Experimentally testable predictions are: 1), phases of individual cells are governed by their intrinsic periods; and 2), efficient synchronization is achieved when the average neurotransmitter concentration would dampen individual oscillators. However, due to the global neurotransmitter oscillation, cells are effectively synchronized. 相似文献