Cooperative, heparan sulfate-dependent cellular uptake of dimeric guanidinoglycosides

Oligoarginine and guanidinium-rich molecular transporters have been shown to facilitate the intracellular delivery of a diverse range of biologically relevant cargos. Several such transporters have been suggested to interact with cell-surface heparan sulfate proteoglycans as part of their cell-entry pathway. Unlike for other guanidinium-rich transporters, the cellular uptake of guanidinoglycosides at nanomolar concentrations is exclusively heparan sulfate dependent. As distinct cells differ in their expression levels and/or the composition of cell-surface heparan sulfate proteoglycans, one might be able to exploit such differences to selectively target certain cell types. To systematically investigate the nature of their cell-surface interactions, monomeric and dimeric guanidinoglycosides were synthesized by using neomycin, paromomycin, and tobramycin as scaffolds. These transporters differ in the number and 3D arrangement of their guanidinium groups. Their cellular uptake was measured by flow cytometry in wild-type and mutant Chinese hamster ovary cells after the corresponding fluorescent streptavidin-phycoerythrin-Cy5 conjugates had been generated. All derivatives showed negligible uptake in mutant cells lacking heparan sulfate. Decreasing the number of guanidinium groups diminished uptake, but the three dimensional arrangement of these groups was less important for cellular delivery. Whereas conjugates prepared with the monomeric carriers showed significantly reduced uptake in mutant cells expressing heparan sulfate chains with altered patterns of sulfation, conjugates prepared with the dimeric guanidinoglycosides could overcome this deficiency and maintain high levels of uptake in such deficient cells. This finding suggests that cellular uptake depends on the valency of the transporter and both the content and arrangement of the sulfate groups on the cell-surface receptors. Competition studies with chemically desulfated or carboxy-reduced heparin derivatives corroborated these observations. Taken together, these findings show that increasing the valency of the transporters retains heparan sulfate specificity and provides reagents that could distinguish different cell types based on the specific composition of their cell-surface heparan sulfate proteoglycans.     

AFM Characterization of Raman Laser-Induced Damage on CdZnTe Crystal Surfaces

Raman laser studies of detector-grade CdZnTe crystals show an increase in intensity of the Te peaks of the Raman spectra even at very low laser powers. In this study, atomic force microscopy (AFM) was used to characterize the extent of damage to the CdZnTe crystal surface following exposure to the Raman laser. AFM images revealed localized surface damage in the areas exposed to the Raman laser beam. Additional studies using conductive-probe AFM techniques provided localized electrical information for the laser-induced Te-rich areas.     

Quantitative and qualitative changes in endogenous nitrogen components along the small intestine of the calf

The aim of this study was to investigate the quantitative and qualitative changes in endogenous protein components along the calf's small intestine. A protein‐free diet and three milk substitutes based on skim milk powder and differing in their protein level (100, 200 and 280 g kg⁻¹ respectively) were prepared. Duodenal, jejunal and ileal digesta were collected through simple cannulae. The flow of non‐specific endogenous amino acids (AAs) determined with the protein‐free diet was 8, 13 and 12 g kg⁻¹ of dry matter intake in the duodenum, jejunum and ileum respectively. The flow of endogenous amino acids in digesta from calves fed diets containing protein was determined using multiple linear regression analysis. It was higher than the non‐specific flow at the duodenum and jejunum owing to additional specific endogenous components. At the ileum, milk proteins were completely digested and the flow of total AAs assayed was equal to the non‐specific flow. Seventy per cent of protein flowing at the jejunum was apparently reabsorbed before the terminal ileum. Endogenous protein at the duodenum resembled a theoretical mixture of proteins from saliva, gastric and pancreatic juices (140, 270 and 590 g kg⁻¹ respectively). Ileal protein losses corresponded to a mixture of proteins from the host and gut bacteria in a 50:50 ratio. By contrast, no theoretical mixture of reference proteins fitted the AA composition of jejunal endogenous protein. © 2000 Society of Chemical Industry     

Identification of Specific and Nonspecific Inhibitors of Bacillus anthracis Type III Pantothenate Kinase (PanK)

Antibiotics with novel mechanisms of action are desperately needed to combat the increasing rates of multidrug-resistant infections. Bacterial pantothenate kinase (PanK) has emerged as a target of interest to cut off the biosynthesis of coenzyme A. Herein we report the results of an in vitro high-throughput screen of over 10 000 small molecules against Bacillus anthracis PanK, as well as a follow-up screen of hits against PanK isolated from Pseudomonas aeruginosa and Burkholderia cenocepacia. Nine hits are structurally categorized and analyzed to set the stage for future drug development.     

Effect of mineralocorticoids on interdialytic weight gain in hemodialysis patients with perdialytic hypotension

Fludrocortisone is recommended in patients with orthostatic hypotension and a benefit has been suggested in hemodialysis patients with severe hypokaliemia. We report 2 patients who suffered from chronic severe perdialytic hypotension resistant to midodrine and who were treated in a long-term period with fludrocortisone. A rise of post-dialytic BP was observed with a decrease of the interdialytic weight gain (IWG). We suggest that the IWG decrease is induced by a lessening of the renin angiotensin aldosterone system that could be less stimulated at the end of the dialysis session because of a better-preserved arterial pressure. The decrease of angiotensin could lessen the feeling of thirst.     

Microstructure and electromigration in copper damascene lines

Grain sizes and crystallographic orientations of Cu were analyzed versus linewidth in damascene Cu interconnects. Pure bamboo lines were not obtained because grain size decreased as linewidth was reduced. Comparison of electromigration results, for wide line Chemical vapor deposition-Cu (3 μm) polycrystalline structure, and narrow lines (0.5 μm) quasi-bamboo structure, provided almost the same activation energy E_a0.65 eV, even though the poor (2 0 0) texture has rotated in the film plane for the narrow damascene lines. These results are in agreement with copper diffusion involving surface diffusion. Besides, even with a polycrystalline crystallographic orientation, PVD-Cu samples showed a better activation energy value E_a=1.02 eV.     

Anti-wear and Friction Reducing Mechanisms of Carbon Nano-onions as Lubricant Additives

Carbon nano-onions have better tribological properties than graphite powder when used as additives dispersed in a poly-alpha-olefin base oil. Carbon nano-onions give a better dispersion in the liquid base oil due to their nanometre-scale size. In particular, the anti-wear efficiency of carbon onions under boundary lubrication and mild wear regime is much better than that of graphite powder. This effect can be attributed to the different structure of the carbon layers in the two species. High-resolution transmission electron microscopy and electron energy-loss spectroscopy were used to characterize the carbon samples, and significant differences in the carbon layer spacing and the density were shown. Wear debris were also observed by the same techniques. In the tribofilm material we depicted new chemical and crystal nano-structures species similar to some microstructures observed in chondrite meteorites, the most interesting one being maghemite iron oxide. The origin of the low friction and wear is still largely unknown.     

Automatic fire pixel detection using image processing: a comparative analysis of rule-based and machine learning-based methods

This paper presents a comparative analysis of state-of-the art image processing-based fire color detection rules and methods in the context of geometrical characteristics measurement of wildland fires. Two new rules and two new detection methods using an intelligent combination of the rules are presented, and their performances are compared with their counterparts. The benchmark is performed on approximately two hundred million fire pixels and seven hundred million non-fire pixels extracted from five hundred wildland images under diverse imaging conditions. The fire pixels are categorized according to fire color and existence of smoke; meanwhile, non-fire pixels are categorized according to the average intensity of the corresponding image. This characterization allows to analyze the performance of each rule by category. It is shown that the performances of the existing rules and methods from the literature are category dependent, and none of them is able to perform equally well on all categories. Meanwhile, a new proposed method based on machine learning techniques and using all the rules as features outperforms existing state-of-the-art techniques in the literature by performing almost equally well on different categories. Thus, this method, promises very interesting developments for the future of metrologic tools for fire detection in unstructured environments.     

Potential dietary sources of ellagic acid and other antioxidants among fruits consumed in Brazil: jabuticaba (Myrciaria jaboticaba (Vell.) Berg)

BACKGROUND: The objectives of this study were (1) to evaluate the content of ellagic acid in fruits consumed by the Brazilian population, including native ones; (2) to further characterize rich sources in relation to ascorbic acid, phenolics contents and in vitro antioxidant capacity; and (3) to study the distribution and effect of ripening stage on ellagitannins content of jabuticaba (Myrciaria jaboticaba). The content of free ellagic acid and ellagic acid derivatives such as ellagitannins was analyzed using high‐performance liquid chromatography (HPLC). RESULTS: Ellagic acid was detected in 10 out of a total of 35 fruits analyzed. The content of free ellagic acid in fruits varied from 0.0028 to 0.085 g kg^?1 (FW) and total ellagic acid varied from 0.215 to 3.11 g kg^?1 (FW). All the seven fruits belonging to the Myrtaceae family evaluated in this study presented high contents of ellagitannins in their composition, with jabuticaba, grumixama and cambuci (all native from Brazil) showing the highest total ellagic acid contents. Jabuticaba, the most consumed in Brazil among those and already adapted to commercial plantations, contained concentrated phenolics compounds, including ellagitannins, in the peel. Anthocyanins (cyanidin derivatives) increased significantly through ripening of jabuticaba and were not present in the pulp or seeds. Samples collected from three different locations during summer, winter and spring had total ellagic contents varying from 1.88 to 3.31 g kg^?1 (FW). The decrease in ellagic acid content with ripening was more accentuated for pulp (eight times) compared to seeds (2.3 times) and peel (2.0 times). CONCLUSION: These results showed the potential of jabuticaba as dietary source of ellagic acid and reinforced consumption of the whole fruit by the population. Copyright © 2011 Society of Chemical Industry