Antigen-specific human T lymphocyte clones: mechanisms of inhibition of proliferative responses by xenoantiserum to human nonpolymorphic HLA-DR antigens

We studied the effects of xenoantiserum to human nonpolymorphic Ia-like antigens upon in vitro antigen-specific T cell proliferative responses in unfractionated PBL populations and at the monoclonal level. Our findings suggest that the xenoantiserum, although it inhibits the antigen-specific response of unfractionated PBL and allospecific T cell clones, does not inhibit the proliferative response to cloned influenza virus immune human T lymphocytes, and therefore may be mediating inhibition by dual mechanisms: direct inhibition of alloantigen recognition and induction of nonspecific suppression. Kinetic differences may explain these phenomena. In cocultivation experiments with a virus-specific clone, the RaIa antiserum appears to induce an OKT3+,8+,4-, radiosensitive regulatory subset of lymphocytes. When adoptively transferred, these induced cells inhibit the TLC response in an antigen-nonspecific and genetically nonrestricted manner. We discuss the various modes and levels of inhibition of antigen-specific proliferation by anti-Ia antisera and their multiple activities.     

Properties of Photosystem II lacking the PsbJ subunit

Photosynthesis Research - Photosystem II (PSII), the oxygen-evolving enzyme, consists of 17 trans-membrane and 3 extrinsic membrane proteins. Other subunits bind to PSII during assembly, like...     

Exploring the temporal variability of a food web using long‐term biomonitoring data

Ecological communities are constantly being reshaped in the face of environmental change and anthropogenic pressures. Yet, how food webs change over time remains poorly understood. Food web science is characterized by a trade‐off between complexity (in terms of the number of species and feeding links) and dynamics. Topological analysis can use complex, highly resolved empirical food web models to explore the architecture of feeding interactions but is limited to a static view, whereas ecosystem models can be dynamic but use highly aggregated food webs. Here, we explore the temporal dynamics of a highly resolved empirical food web over a time period of 18 years, using the German Bight fish and benthic epifauna community as our case study. We relied on long‐term monitoring ecosystem surveys (from 1998 to 2015) to build a metaweb, i.e. the meta food web containing all species recorded over the time span of our study. We then combined time series of species abundances with topological network analysis to construct annual food web snapshots. We developed a new approach, ‘node‐weighted’ food web metrics by including species abundances to represent the temporal dynamics of food web structure, focusing on generality and vulnerability. Our results suggest that structural food web properties change through time; however, binary food web structural properties may not be as temporally variable as the underlying changes in species composition. Further, the node‐weighted metrics enabled us to detect that food web structure was influenced by changes in species composition during the first half of the time series and more strongly by changes in species dominance during the second half. Our results demonstrate how ecosystem surveys can be used to monitor temporal changes in food web structure, which are important ecosystem indicators for building marine management and conservation plans.     

Validation of a video-based motion analysis technique in 3-D dynamic scapular kinematic measurements

BackgroundCurrent non-invasive 3-D scapular kinematic measurement techniques such as electromagnetic tracking are subjected to restrictions of wired sensors and limited capture space. Video-based motion analysis provides greater freedom with relatively less movement restriction. However, video-based motion analysis was rarely used in and not validated for scapular kinematics.MethodsScapular kinematics of five subjects performing abduction, scaption, and internal/external rotation was captured simultaneously with video-based motion analysis and dynamic stereo X-ray, a gold standard for tracking scapular movements. The data from video-based motion analysis was correlated with the data from dynamic stereo X-ray for validity evaluation.FindingsStrong and significant correlations were identified in scapular protraction/retraction and medial/lateral rotation during abduction and scaption, and scapular medial/lateral rotation and anterior/posterior tilt during internal/external rotation.InterpretationVideo-based motion analysis is valid for evaluating a single subject's scapular movement pattern in protraction/retraction during abduction and scaption, and medial/lateral-rotation during internal/external rotation. Anterior/posterior-tilt during abduction and scaption should be investigated with caution. Video motion analysis is also valid for evaluating group average of scapular kinematics except for protraction/retraction during internal/external rotation. While acknowledging the inherent limitations, video-based motion analysis is an appropriate technique for tracking scapular kinematics.     

Ku80 facilitates chromatin binding of the telomere binding protein,TRF2

The Ku70/80 heterodimer is central to non-homologous end joining repair of DNA double-strand breaks and the Ku80 gene appears to be essential for human but not rodent cell survival. The Ku70/80 heterodimer is located at telomeres but its precise function in telomere maintenance is not known. In order to examine the role of Ku80 beyond DNA repair in more detail, we have taken a knockdown approach using a human fibroblast strain. Following targeted Ku80 knockdown, telomere defects are observed and the steady state levels of the TRF2 protein are reduced. Inhibitor studies indicate that this loss of TRF2 is mediated by the proteasome and degradation of TRF2 following Ku depletion appears to involve a decrease in chromatin binding of TRF2, suggesting that the Ku heterodimer enhances TRF2 chromatin association and that non-chromatin bound TRF2 is targeted to the proteasome.Key words: Ku80, TRF2, chromatin, telomere, fibroblast     

Conversion of arginine into ornithine by advanced glycation in senescent human collagen and lens crystallins

Long lived proteins undergo age-related postsynthetic modifications that destabilize them by altering their conformation, charge, and helicity, thereby enhancing their resistance toward proteolysis and propensity to aggregate. The unexpected finding of substantial amounts of ornithine, the nonprotein amino acid, and decarbamidation product of arginine in acid hydrolysates of lens crystallins and skin collagen led us to investigate its source and mechanism of formation. In order to exclude ornithine formation as an artifact of acid hydrolysis, proteins were reductively alkylated with formaldehyde to convert ornithine to dimethyl-ornithine. The proteins were assayed for carboxymethyl-ornithine and glycated ornithine ("furornithine") by liquid chromatography coupled to electrospray ionization mass spectrometry. Ornithine in acid hydrolysates of human lens and skin proteins increased from 1 to 15 nmol/mg protein from ages 10 to 90 years, whereas dimethyl-ornithine increased from 0.5 to 15 and from 0 to 5 nmol/mg protein, respectively. Carboxymethyl-ornithine and furornithine increased with age in lens and skin from approximately 0 to 60 and 0 to 180 pmol/mg protein, respectively. In collagen, ornithine was elevated above levels of nondiabetic controls only when both diabetes and end stage renal disease were present. The age-related increase of these modifications provides evidence for substantial in vivo formation of ornithine in aging human tissue proteins. The mechanism of ornithine formation is not known, but data suggest that arginine-derived advanced glycation end products might serve as precursors for the in vivo conversion of ornithine from arginine.     

Extension of replicative lifespan in WI-38 human fibroblasts by dexamethasone treatment is accompanied by suppression of p21 Waf1/Cip1/Sdi1 levels

Numerous studies have shown that supplementation of the growth medium of human fibroblasts with dexamethasone at physiologic concentrations extends replicative lifespan up to 30%. While this extension of lifespan has been used to probe various aspects of the senescent phenotype, no mechanism for the increased lifespan of human fibroblasts grown in the presence of dexamethasone has ever been identified. In the present study we present evidence that the extended lifespan of human lung fibroblasts (WI-38 cells) that occurs when these cells are maintained in culture medium supplemented with dexamethasone is accompanied by a suppression of p21(Waf1/Cip1/Sdi1) levels, which normally increase as these cells enter senescence, while p16(INK4a) levels are unaffected. These results suggest that the delay of senescence in cultures grown in the presence of dexamethasone is due to a suppression of the senescence related increase in p21(Waf1/Cip1/Sdi1). These results are consistent with models of replicative senescence in which p53 and p21(Waf1/Cip1/Sdi1) play a role in the establishment of the senescent arrest.