Photoinhibition of Photosystem I damages both reaction centre proteins PSI-A and PSI-B and acceptor-side located small Photosystem I polypeptides

Photoinhibition of Photosystem I at chilling temperatures was investigated. Illumination of barley and cucumber leaves at 4°C induced a lowered Photosystem I activity. In barley, the reaction centre proteins PSI-A and PSI-B were both partially degraded as was the nuclear-encoded PSI-D polypeptide. Barley leaves infiltrated with KCN to increase oxidative stress, showed increased photoinhibition of Photosystem I, including reduced photochemical activity and marked degradation of several Photosystem I polypeptides. The most rapid and pronounced degradation was found in the PSI-D and PSI-E polypeptides exposed at the Photosystem I acceptor side. The PSI-A, -B, -C, -G, -H, -K and -L polypeptides were less extensively damaged. No damage of the lumenally oriented PSI-F and -N polypeptides was detected. The elevated photoinhibition of Photosystem I seen in KCN treated barley is most likely induced by a combination of increased active oxygen due to inhibited scavenging and increased accumulation of reducing power due to inhibition of the Calvin cycle. In barley, photo-inactivation of Photosystem I closely followed the degradation of PSI-A and PSI-B. Illumination of cucumber resulted in a pronounced loss of activity and appearance of specific PSI-A and PSI-B degradation products whereas the total PSI-A/B degradation was small. The PSI-A/B degradation identified in barley is interpreted to reflect a physiologically relevant process being part of a repair cycle, whereas the much smaller PSI-A/B degradation observed in cucumber is interpreted to represent an irreversible damage induced far below the temperature tolerance for cucumber.     

Nitric oxide‐mediated dispersal in single‐ and multi‐species biofilms of clinically and industrially relevant microorganisms

Strategies to induce biofilm dispersal are of interest due to their potential to prevent biofilm formation and biofilm‐related infections. Nitric oxide (NO), an important messenger molecule in biological systems, was previously identified as a signal for dispersal in biofilms of the model organism Pseudomonas aeruginosa. In the present study, the use of NO as an anti‐biofilm agent more broadly was assessed. Various NO donors, at concentrations estimated to generate NO levels in the picomolar and low nanomolar range, were tested on single‐species biofilms of relevant microorganisms and on multi‐species biofilms from water distribution and treatment systems. Nitric oxide‐induced dispersal was observed in all biofilms assessed, and the average reduction of total biofilm surface was 63%. Moreover, biofilms exposed to low doses of NO were more susceptible to antimicrobial treatments than untreated biofilms. For example, the efficacy of conventional chlorine treatments at removing multi‐species biofilms from water systems was increased by 20‐fold in biofilms treated with NO compared with untreated biofilms. These data suggest that combined treatments with NO may allow for novel and improved strategies to control biofilms and have widespread applications in many environmental, industrial and clinical settings.     

A G protein-coupled receptor-like module regulates cellulose synthase secretion from the endomembrane system in Arabidopsis

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NickFects,Phosphorylated Derivatives of Transportan 10 for Cellular Delivery of Oligonucleotides

Oligonucleotide-based gene regulation has a high potential in gene therapy, but the plasma membrane is impermeable for nucleic acid polymers and, consequently, an efficient and non-toxic transfection agent is needed for their delivery into the cell. In this study we present a novel series, NickFects, of chemically modified TP10 peptide-based delivery vectors used for the cellular delivery of single-stranded oligonucleotides. These carriers, obtained by replacement of Ile8 by threonine in stearyl-TP10 and by modifying of tyrosine and/or threonine, respectively, by phosphorylation formed 300–500 nm in size peptide:oligonucleotide nanocomplexes with negative surface charges. The highest splice-correcting effect was obtained when phosphorotiate 2′-O-methyl oligonucleotides were transduced into cells by NickFect1 (NF1) or NickFect2 (NF2). In addition, we also show how a small modification (one or two negative charges) in peptide sequence can affect its ability to deliver ONs into cells and increase their potency in the splicing redirection assay. Our studies demonstrate that NF1 and NF2 have higher transfection efficacy for oligonucleotides as compared to the most commonly used transfection agent Lipofectamine™ 2000 and lead to higher biological response in cells.     

Humoral and cellular immune reactions against tumor cells in patients with urinary bladder carcinoma

Serum IgG fractions from a large and homogeneous group of patients with transitional cell carcinoma of the urinary bladder (TCC) were tested for their capacity to induce antibody-dependent cellular cytotoxicity (ADCC) with lymphocytes from healthy donors against a TCC-derived target cell and one derived from adenocarcinoma of the colon. Both targets have previously been shown to be of comparable susceptibility to cell-mediated lysis in vitro. Some of the IgG preparations showed strong and dose-dependent ADCC against either one or both targets, while others gave weak reactions or none at all. Similar results were obtained with IgG from a matched group of patients with prostatic carcinoma who were used as clinical controls (CC). In parallel experiments, lymphocytes taken from the two donor groups at the same time as the serum samples were tested for their direct cytotoxicity (CMC) against the two targets. CMC gave similar results to ADCC. The differences in cytotoxicity displayed by either IgG or lymphocytes from individual donors were analysed statistically, using nonparametric statistics. To avoid introducing bias due to arbitrary data selection, the entire set of results, comprising both high and low reactors, was included in the statistical assessment. ADCC of the TCC donors' IgG against the TCC target was significantly stronger than against the colon carcinoma and also significantly stronger than that of the control donors. Similarly, the TCC patients' lymphocytes displayed a significantly higher CMC against the TCC target than against the control targets. This was not seen when the lymphocytes from the patients with prostatic carcinoma were tested. When CMC and ADCC of individual donors were compared, a statistically significant correlation between these activities was seen in three of the four donor/target combinations. These results support earlier findings and suggest that a significant fraction of both the disease-related and the 'non-selective' CMC (NK) displayed by cancer patients lymphocytes against allogeneic tumor cells in vitro reflects antibody-dependent reactions.     

Non-stationary Response of Tree Growth to Climate Trends Along the Arctic Margin

Ecosystems - Climate change modulates cold-marginal forest ecosystems through changing growth constraints. Understanding spatiotemporal variations in climate–growth relationships is essential...     

Characterization of Lymphocyte Fibronectin

In vitrocultured “activated” peripheral blood lymphocytes and T-cell lines synthesized a high-molecular-weight gelatin binding molecule (MW 500 kDa), whereas resting lymphocytes showed poor or negligible synthesis of the same component. Concanavalin A-mediated anchorage of the lymphocytes to a substratum potentiated synthesis of the high-molecular-weight molecule. Western blotting of the gelatin-binding lymphocyte molecule demonstrated reactivity with antibodies specific for human fibronectin. Furthermore, immunocytochemistry showed reactivity of anti-fibronectin antibodies with T-lymphocytes at the single-cell level. The lymphocyte-derived fibronectin was preferentially cell associated and relatively small amounts were present in the culture medium. RT-PCR of total RNA from CD4⁺T-cells and the lymphoid T-cell line MOLT-4 showed that the most abundant species of fibronectin mRNA lacked the entire III CS exon encoding the α₄β₁binding region LDV. Amplification of the III CS region from other T-cell lines revealed that these cells expressed several fibronectin mRNA isoforms most of which were lacking the LDV coding sequence. In conclusion, synthesis of fibronectin is demonstrated to occur in T-lymphocytes and to be regulated by signals which activate the cells.     

Lack of correlation between the rate of cholesterol biosynthesis and the activity of 3-hydroxy-3-methylgutaryl coenzyme A reductase in rats and in fibroblasts treated with ML-236B

The phospholipase-2 inhibitor, p-bromophenacyl bromide, has been shown to inhibit strongly the elongation of endogenous fatty acids in preparations of brain mitochondria and microsomes. On the other hand, it does not inhibit the elongation of added palmitic or linoleic acids. The implication is that the normal first step in alteration of membrane lipid fatty acids is their release to other membrane-bound enzyme systems by a membrane-bound phospholipase A.     

Factors controlling the in vitro growth pattern of human microvascular endothelial cells

Monolayer cultures of endothelial cells of human dermal microvascular origin were exposed to a variety of culture conditions and in vitro differentiation of the cells assessed by light and electron microscopic examination. Restoration of a cytologic and fine structural appearance which resembled most closely that present in vivo was possible by raising the intracellular cAMP level. These cells formed junctional complexes seen in uncontracted microvessels and specialized attachment sites at their basal cell membrane, contained a complex network of bundled micro- and intermediate filaments and numerous Weibel-Palade bodies and accumulated electron-opaque deposits between the cells and the culture dish surface.     

Life cycle assessment of base–load heat sources for district heating system options

Purpose  

There has been an increased interest in utilizing renewable energy sources in district heating systems. District heating systems are centralized systems that provide heat for residential and commercial buildings in a community. While various renewable and conventional energy sources can be used in such systems, many stakeholders are interested in choosing the feasible option with the least environmental impacts. This paper evaluates and compares environmental burdens of alternative energy source options for the base–load of a district heating center in Vancouver, British Columbia (BC) using the life cycle assessment method. The considered energy sources include natural gas, wood pellet, sewer heat, and ground heat.