Anti-PlGF inhibits growth of VEGF(R)-inhibitor-resistant tumors without affecting healthy vessels

Novel antiangiogenic strategies with complementary mechanisms are needed to maximize efficacy and minimize resistance to current angiogenesis inhibitors. We explored the therapeutic potential and mechanisms of alphaPlGF, an antibody against placental growth factor (PlGF), a VEGF homolog, which regulates the angiogenic switch in disease, but not in health. alphaPlGF inhibited growth and metastasis of various tumors, including those resistant to VEGF(R) inhibitors (VEGF(R)Is), and enhanced the efficacy of chemotherapy and VEGF(R)Is. alphaPlGF inhibited angiogenesis, lymphangiogenesis, and tumor cell motility. Distinct from VEGF(R)Is, alphaPlGF prevented infiltration of angiogenic macrophages and severe tumor hypoxia, and thus, did not switch on the angiogenic rescue program responsible for resistance to VEGF(R)Is. Moreover, it did not cause or enhance VEGF(R)I-related side effects. The efficacy and safety of alphaPlGF, its pleiotropic and complementary mechanism to VEGF(R)Is, and the negligible induction of an angiogenic rescue program suggest that alphaPlGF may constitute a novel approach for cancer treatment.     

Plant-insect communities and predator-prey ratios in field margin strips,adjacent crop fields,and fallows

The management of field margin strips for the enhancement of biodiversity of plant-insect communities and natural-enemy populations was studied on experimental farms near Göttingen (Germany). Young and old, sown and naturally developed field margin strips were compared and differences to large fallows established. The five types of field margin strips (around cereal fields) were: (1, 2) 1- or 6-year-old naturally developed strips, (3) strips sown with a Phacelia mixture, (4) strips sown with a mixture of 19 wild flower species, and (5) strips sown with winter wheat or oat as a control. The naturally developed vegetation of the field margin strips was dominated by aggressive weeds, presumably due to the intensive farming practices and the fertile soils. Cirsium arvense populations decreased, while Elymus repens populations increased with age of habitat. Sowings were suitable to suppress these aggressive weeds. Potted plants of mugwort (Artemisia vulgaris) and red clover (Trifolium pratense) were exposed in the field margin strips to study arthropod colonization of these experimentally standardized plant patches. Arthropod species richness did not differ between field margin types, reflecting the overall similarity in floristic diversity, but sprayed and strip-free edges of cereal fields had a reduced diversity. Dispersal of insect populations of red clover into the cereal fields decreased with increasing distance, but benefited from adjacent field margin strips. Populations of predators (mainly spiders) as well as predator-prey ratios were significantly larger in 6-year-old than in 1-year-old strips emphasizing the importance of habitat age for natural enemies and possible biological control. Predator-prey ratios were also higher on old than young fallows. Large fallows had greater predator-prey ratios than small field margin strips emphasizing the trophic-level hypothesis of island biogeography in that the relative importance of natural enemies increased with habitat area. Insect species richness was only marginally influenced by area and not by age. As species richness of predators did not increase with area and age, species diversity and the possible biological-control function did not covary.     

Influence of morphology and rheology on the production characteristics of the basidiomycete Cyathus striatus

The influence of the pellet morphology of the basidiomycete Cyathus striatus on the production of the antibiotics striatals A, B, and C was investigated. The main operating parameters in fermenters of different sizes were the tip speed and the volumetric power input. Different methods were developed for quantification of morphological characteristics. The apparent viscosity of the suspension was measured with a cylinder rheometer. Sediment density was measured with a sedimentation apparatus. Particle size distributions were recorded with an image analysis system. By means of the presented measuring methods, morphological characteristics could be determined and enabled an early assessment of the fermentation course and the antibiotics production. During the exponential growth phase of the fungus the relative sediment height correlated with the biomass concentration. The pellet morphology at this stage influenced the later production of striatals. The yield of the striatals was markedly influenced by pellet size and sediment density. Since these morphological characteristics determine the rheological properties of the culture the measurement of the apparent viscosity of the culture in the production phase allowed predictions of the production yield.     

Interactor-mediated nuclear translocation and retention of protein phosphatase-1

Protein Ser/Thr phosphatase-1 (PP1) is a ubiquitous eukaryotic enzyme that controls numerous cellular processes by the dephosphorylation of key regulatory proteins. PP1 is expressed in various cellular compartments but is most abundant in the nucleus. We have examined the determinants for the nuclear localization of enhanced green fluorescent protein-tagged PP1 in COS1 cells. Our studies show that PP1gamma(1) does not contain a functional nuclear localization signal and that its nuclear accumulation does not require Sds22, which has previously been implicated in the nuclear accumulation of PP1 in yeast (Peggie, M. W., MacKelvie, S. H., Bloecher, A., Knatko, E. V., Tatchell, K., and Stark, M. J. R. (2002) J. Cell Sci. 115, 195-206). However, the nuclear targeting of PP1 isoforms was alleviated by the mutation of their binding sites for proteins that interact via an RVXF motif. Moreover, one of the mutants with a cytoplasmic accumulation and decreased affinity for RVXF motifs (PP1gamma(1)-F257A) could be re-targeted to the nucleus by the overexpression of nuclear interactors (NIPP1 (nuclear inhibitor of PP1) and PNUTS (PP1 nuclear targeting subunit)) with a functional RVXF motif. Also, the addition of a synthetic RVXF-containing peptide to permeabilized cells resulted in the loss of nuclear enhanced green fluorescent protein-PP1gamma(1). Finally, NIPP1(-/-) mouse embryos showed a nuclear hyperphosphorylation on threonine, consistent with a role for NIPP1 in the nuclear targeting and/or retention of PP1. Our data suggest that both the nuclear translocation and the nuclear retention of PP1 depend on its binding to interactors with an RVXF motif.     

Exercise flow-volume loops in prepubescent aerobically trained children.

We studied mechanical ventilatory constraints in 13 aerobically trained (Tr) and 11 untrained (UT) prepubescent children by plotting the exercise flow-volume (F-V) loops within the maximal F-V loop (MFVL) measured at rest. The MFVL allowed to determine forced vital capacity (FVC) and maximal expiratory flows. Expiratory and inspiratory reserve volumes relative to FVC (ERV/FVC and IRV/FVC, respectively) were measured during a progressive exercise test until exhaustion. Breathing reserve (BR) and expiratory flow limitation (expFL), expressed in percentage of tidal volume (V(T)) and defined as the part of the tidal breath meeting the boundary of the MFVL, were measured. Higher FVC and maximal expiratory flows were found in Tr than UT (P < 0.05) at rest. Our results have shown that during exercise, excepting one subject, all Tr regulated their V(T) within FVC similarly during exercise, by breathing at low lung volume at the beginning of exercise followed breathing at high lung volume at strenuous exercise. In UT, ERV/FVC and IRV/FVC were regulated during exercise in many ways. The proportion of children who presented an expFL was nearly the same in both groups (approximately 70% with a range of 14 to 65% of V(T)), and no significant difference was found during exercise concerning expFL. However, higher ventilation (V(E)), ERV/FVC, and dyspnea associated with lower BR, IRV/FVC, and SaO2 were reported at peak power in Tr than UT (P < 0.05). These results suggest that, because of their higher Ve level, trained children presented higher ventilatory constraints than untrained. These may influence negatively the SaO2 level and dyspnea during strenuous exercise.     

Phosphoproteomics data classify hematological cancer cell lines according to tumor type and sensitivity to kinase inhibitors

Background

Tumor classification based on their predicted responses to kinase inhibitors is a major goal for advancing targeted personalized therapies. Here, we used a phosphoproteomic approach to investigate biological heterogeneity across hematological cancer cell lines including acute myeloid leukemia, lymphoma, and multiple myeloma.

Results

Mass spectrometry was used to quantify 2,000 phosphorylation sites across three acute myeloid leukemia, three lymphoma, and three multiple myeloma cell lines in six biological replicates. The intensities of the phosphorylation sites grouped these cancer cell lines according to their tumor type. In addition, a phosphoproteomic analysis of seven acute myeloid leukemia cell lines revealed a battery of phosphorylation sites whose combined intensities correlated with the growth-inhibitory responses to three kinase inhibitors with remarkable correlation coefficients and fold changes (> 100 between the most resistant and sensitive cells). Modeling based on regression analysis indicated that a subset of phosphorylation sites could be used to predict response to the tested drugs. Quantitative analysis of phosphorylation motifs indicated that resistant and sensitive cells differed in their patterns of kinase activities, but, interestingly, phosphorylations correlating with responses were not on members of the pathway being targeted; instead, these mainly were on parallel kinase pathways.

Conclusion

This study reveals that the information on kinase activation encoded in phosphoproteomics data correlates remarkably well with the phenotypic responses of cancer cells to compounds that target kinase signaling and could be useful for the identification of novel markers of resistance or sensitivity to drugs that target the signaling network.     

Fibrinogen alpha chain O-glycopeptides as possible markers of urinary tract infection

Urinary tract infection (UTI) is the most common bacterial infection leading to substantial morbidity and considerable health care expenditures across all ages. Here we present an exploratory UPLC-MS study of human urine in the context of febrile, complicated urinary tract infection aimed to reveal and identify possible markers of a host response on infection. A UPLC-MS based workflow, taking advantage of Ultra High Resolution (UHR) Qq-ToF-MS, and multivariate data handling were applied to a carefully selected group of 39 subjects with culture-confirmed febrile Escherichia coli UTI. Using a combination of unsupervised and supervised multivariate modeling we have pinpointed a number of peptides specific for UTI. An unequivocal structural identification of these peptides, as O-glycosylated fragments of the human fibrinogen alpha 1 chain, required MS2 and MS3 experiments on two different MS platforms: ESI-UHR-Qq-ToF and ESI-ion trap, a blast search and, finally, confirmation was achieved by matching experimental tandem mass spectra with those of custom synthesized candidate-peptides.In conclusion, exploiting non-targeted UPLC-MS based approach for the investigation of UTI related changes in urine, we have identified and structurally characterized unique O-glycopeptides, which are, to our knowledge, the first demonstration of O-glycosylation of human fibrinogen alpha 1-chain.     

Fungal dual-domain LysM effectors undergo chitin-induced intermolecular,and not intramolecular,dimerization

Chitin is a homopolymer of β-(1,4)-linked N-acetyl-D-glucosamine (GlcNAc) and a major structural component of fungal cell walls. In plants, chitin acts as a microbe-associated molecular pattern (MAMP) that is recognized by lysin motif (LysM)-containing plant cell surface-localized pattern recognition receptors (PRRs) that activate a plethora of downstream immune responses. To deregulate chitin-induced plant immunity and successfully establish infection, many fungal pathogens secrete LysM domain-containing effector proteins during host colonization. The LysM effector Ecp6 from the tomato (Solanum lycopersicum) leaf mold fungus Cladosporium fulvum can outcompete plant PRRs for chitin binding because two of its three LysM domains cooperate to form a composite groove with ultra-high (pM) chitin-binding affinity. However, most functionally characterized LysM effectors contain only two LysMs, including Magnaporthe oryzae MoSlp1, Verticillium dahliae Vd2LysM, and Colletotrichum higginsianum ChElp1 and ChElp2. Here, we performed modeling, structural, and functional analyses to investigate whether such dual-domain LysM effectors can also form ultra-high chitin-binding affinity grooves through intramolecular LysM dimerization. However, our study suggests that intramolecular LysM dimerization does not occur. Rather, our data support the occurrence of intermolecular LysM dimerization for these effectors, associated with a substantially lower chitin binding affinity than monitored for Ecp6. Interestingly, the intermolecular LysM dimerization allows for the formation of polymeric complexes in the presence of chitin. Possibly, such polymers may precipitate at infection sites to eliminate chitin oligomers, and thus suppress the activation of chitin-induced plant immunity.

Fungal LysM effectors composed of two LysM domains bind chitin via intermolecular LysM dimerization, leading to polymers that may precipitate to eliminate chitin from infection sites to prevent the activation of host immune receptors.     

Circulating Lipoproteins Are a Crucial Component of Host Defense against Invasive Salmonella typhimurium Infection

Background

Circulating lipoproteins improve the outcome of severe Gram-negative infections through neutralizing lipopolysaccharides (LPS), thus inhibiting the release of proinflammatory cytokines.

Methods/Principal Findings

Low density lipoprotein receptor deficient (LDLR−/−) mice, with a 7-fold increase in LDL, are resistant against infection with Salmonella typhimurium (survival 100% vs 5%, p<0.001), and 100 to 1000-fold lower bacterial burden in the organs, compared with LDLR+/+ mice. Protection was not due to differences in cytokine production, phagocytosis, and killing of Salmonella organisms. The differences were caused by the excess of lipoproteins, as hyperlipoproteinemic ApoE−/− mice were also highly resistant to Salmonella infection. Lipoproteins protect against infection by interfering with the binding of Salmonella to host cells, and preventing organ invasion. This leads to an altered biodistribution of the microorganisms during the first hours of infection: after intravenous injection of Salmonella into LDLR+/+ mice, the bacteria invaded the liver and spleen within 30 minutes of infection. In contrast, in LDLR−/− mice, Salmonella remained constrained to the circulation from where they were efficiently cleared, with decreased organ invasion.

Conclusions

plasma lipoproteins are a potent host defense mechanism against invasive Salmonella infection, by blocking adhesion of Salmonella to the host cells and subsequent tissue invasion.