Aggregated Multicast – A Comparative Study

Though IP multicast is resource efficient in delivering data to a group of members simultaneously, it suffers from scalability problem with the number of concurrently active multicast groups because it requires a router to keep forwarding state for every multicast tree passing through it. To solve this state scalability problem, we proposed a scheme, called aggregated multicast. The key idea is that multiple groups are forced to share a single delivery tree. In our earlier work, we introduced the basic concept of aggregated multicast and presented some initial results to show that multicast state can be reduced. In this paper, we develop a more quantitative assessment of the cost/benefit trade-offs. We propose an algorithm to assign multicast groups to delivery trees with controllable cost and introduce metrics to measure multicast state and tree management overhead for multicast schemes. We then compare aggregated multicast with conventional multicast schemes, such as source specific tree scheme and shared tree scheme. Our extensive simulations show that aggregated multicast can achieve significant routing state and tree management overhead reduction while containing the expense of extra resources (bandwidth waste and tunnelling overhead). We conclude that aggregated multicast is a very cost-effective and promising direction for scalable transit domain multicast provisioning.     

Capsaicin pretreatment attenuates LPS-induced hypothermia through TRPV1-independent mechanisms in chicken

It has been demonstrated that chicken TRPV1 (transient receptor potential vanilloid of subtype-1) is insensitive to capsaicin (CAP), and therefore, a chicken model is suitable to analyze the CAP-sensitive TRPV1-independent pathway. We elucidated here the possible involvement of the pathway in hypothermia induced by bacterial endotoxin (lipopolysaccharide, LPS) in chickens. Chicks were pretreated with CAP (10 mg/kg, iv) at 1, 2 and 3 days of age to desensitize them towards the CAP-sensitive pathway. An intravenous injection of LPS in 4-day-old chicks caused progressive hypothermia, ending with collapse and 78% mortality within 12 h after injection. The CAP pretreatment rescued the LPS-induced endotoxin shock and hypothermia in chicks. LPS-induced iNOS expression as well as NO production in liver and lung was suppressed by CAP pretreatment. CAP pretreatment also attenuated hypothermia due to exposure of chicks to cold ambient temperature. These findings suggest that a CAP-sensitive TRPV1-independent pathway may be involved in pathophysiological hypothermic reactions through the mediation of NO in chickens.     

Identification of the oriI-binding site of poliovirus 3C protein by nuclear magnetic resonance spectroscopy

Replication of picornaviral genomes requires recognition of at least three cis-acting replication elements: oriL, oriI, and oriR. Although these elements lack an obvious consensus sequence or structure, they are all recognized by the virus-encoded 3C protein. We have studied the poliovirus 3C-oriI interaction in order to begin to decipher the code of RNA recognition by picornaviral 3C proteins. oriI is a stem-loop structure that serves as the template for uridylylation of the peptide primer VPg by the viral RNA-dependent RNA polymerase. In this report, we have used nuclear magnetic resonance (NMR) techniques to study 3C alone and in complex with two single-stranded RNA oligonucleotides derived from the oriI stem. The (1)H-(15)N spectra of 3C recorded in the presence of these RNAs revealed site-specific chemical shift perturbations. Residues that exhibit significant perturbations are primarily localized in the amino terminus and in a highly conserved loop between residues 81 and 89. In general, the RNA-binding site defined in this study is consistent with predictions based on biochemical and mutagenesis studies. Although some residues implicated in RNA binding by previous studies are perturbed in the 3C-RNA complex reported here, many are unique. These studies provide unique site-specific insight into residues of 3C that interact with RNA and set the stage for detailed structural investigation of the 3C-RNA complex by NMR. Interpretation of our results in the context of an intact oriI provides insight into the architecture of the picornavirus VPg uridylylation complex.     

Synthesis of bis-diosgenin pyrazine dimers: new cephalostatin analogs

A convenient synthesis for bis-diosgenin pyrazine dimers, cephalostatin analogues is reported. These symmetrical dimeric steroid-pyrazines were obtained by the classical condensation of -amino ketones, the most efficient method for pyrazine ring construction.     

Crosslinking of alginic acid/chitosan matrices using polycarboxylic acids and their utilization for sodium diclofenac release

Three different ratios of alginic acid/chitosan matrices of ratios 3/1, 1/1 and 1/2, respectively, were crosslinked in their dry state using citric acid (CA)/sodium hypophosphite (SHP) at different conditions controlling the crosslinking process such as citric acid concentration, citric acid/sodium hypophosphite molar ratio as well as time and temperature of reaction. Results indicate that such matrices were crosslinked efficiently on curing at 180 °C for 9 min in presence of CA/SHP ratio 1 and the citric acid concentration of 0.6 based on the weight of any matrices. The crosslinked matrices were characterized by investigating their swelling properties, FT-IR and thermalgravimetric analysis. Furthermore, such crosslinked matrices were tested as drug release for sodium diclofenac.     

Solution structure and model membrane interactions of temporins-SH, antimicrobial peptides from amphibian skin. A NMR spectroscopy and differential scanning calorimetry study

Temporin-SHa and temporin-SHc are 13 residue long antimicrobial peptides from frog skin that have similar sequences but differ markedly in their membrane-damaging properties. Temporin-SHa contains a single basic lysine residue and has a unique antimicrobial spectrum of action among temporins, being very potent against Gram-positive and Gram-negative bacteria, yeasts, fungi, and protozoa. Temporin-SHc, which contains a single basic histidine residue, is inactive against Gram-negative bacteria, has a reduced efficacy against Gram-positive bacteria, but is still active against yeasts and fungi. Temporin-SHb, with no basic residue, has no antimicrobial activity. The three-dimensional structures of the peptides bound to SDS micelles were analyzed by CD and NMR spectroscopy combined with restrained molecular dynamics calculations. The peptides adopt well-defined amphipathic alpha-helical structures extending from residue 3 to residue 12, when bound to SDS micelles. The structures are stabilized by extensive interactions between aliphatic and aromatic side chains on the nonpolar face. Relaxation enhancements caused by paramagnetic probes showed that the peptides adopt nearly parallel orientations to the micelle surface and do not deeply penetrate into the micelle. The interaction of the peptides with model membranes was investigated by differential scanning calorimetry on anionic and zwitterionic multilamellar vesicles and membrane-permeabilization assays on calcein-loaded large unilamellar vesicles. Calorimetric data indicated that both temporin-SHa and -SHc reside at the hydrocarbon core-water interface of the anionic lipid bilayer but interact with anionic bilayers in a very different manner. This suggests that the charge-induced activity of temporins-SH for bacterial cells is due to changes in the membrane-disturbing mechanism of the bound peptides.     

Coronary nitric oxide production controls cardiac substrate metabolism during pregnancy in the dog

The aim of this study was to examine the role of nitric oxide (NO) in the control of cardiac metabolism at 60 days of pregnancy (P60) in the dog. There was a basal increase in diastolic coronary blood flow during pregnancy and a statistically significant increase in cardiac output (55 +/- 4%) and in cardiac NOx production (44 +/- 4 to 59 +/- 3 nmol/min, P < 0.05). Immunohistochemistry of the left ventricle showed an increase in endothelial nitric oxide synthase staining in the endothelial cells at P60. NO-dependent coronary vasodilation (Bezold-Jarisch reflex) was increased by 20% and blocked by N(G)-nitro-l-arginine methyl ester (l-NAME). Isotopically labeled substrates were infused to measure oleate, glucose uptake, and oxidation. Glucose oxidation was not significantly different in P60 hearts (5.4 +/- 0.5 vs. 6.2 +/- 0.4 micromol/min) but greatly increased in response to l-NAME injection (to 19.9 +/- 0.9 micromol/min, P < 0.05). Free fatty acid (FFA) oxidation was increased in P60 (from 5.3 +/- 0.6 to 10.4 +/- 0.5 micromol/min, P < 0.05) and decreased in response to l-NAME (to 4.5 +/- 0.5 micromol/min, P < 0.05). There was an increased oxidation of FFA for ATP production but no change in the respiratory quotient during pregnancy. Genes associated with glucose and glycogen metabolism were downregulated, whereas genes involved in FFA oxidation were elevated. The acute inhibition of NO shifts the heart away from FFA and toward glucose metabolism despite the downregulation of the carbohydrate oxidative pathway. The increase in endothelium-derived NO during pregnancy results in a tonic inhibition of glucose oxidation and reliance on FFA uptake and oxidation to support ATP synthesis in conjunction with upregulation of FFA metabolic enzymes.     

Correlation Between Bacterial Hemoglobin and Carbon Sources: Their Effect on Copper Uptake by Transformed E. coli Strain αDH5

Escherichia coli αDH5 harboring Vitreoscilla hemoglobin gene, vgb (Escherichia:pUC8:16), and control strains (parental and vgb-lacking transformed strains, E. coli:pUC9) were employed for studying the potential ability of copper uptake. The strains were grown on a minimal medium, 5XM9, that contained different carbon (mannitol, fructose, lactose, and maltose) and nitrogen (casein and yeast extract) sources. A clear type–response relationship between the level of copper uptake and the type of carbon and nitrogen sources was observed. Copper uptake ability was better for vgb-bearing cells grown on either mannitol, lactose, fructose, or maltose as carbon source and casein as well as yeast extract as nitrogen source compared with the control medium. The increase in copper uptake was in the following order: parental cells, then pUC9-bearing cells, then VHb-expressing cells.     

Cellulose binding domains of a Phytophthora cell wall protein are novel pathogen-associated molecular patterns

The cellulose binding elicitor lectin (CBEL) from Phytophthora parasitica nicotianae contains two cellulose binding domains (CBDs) belonging to the Carbohydrate Binding Module1 family, which is found almost exclusively in fungi. The mechanism by which CBEL is perceived by the host plant remains unknown. The role of CBDs in eliciting activity was investigated using modified versions of the protein produced in Escherichia coli or synthesized in planta through the potato virus X expression system. Recombinant CBEL produced by E. coli elicited necrotic lesions and defense gene expression when injected into tobacco (Nicotiana tabacum) leaves. CBEL production in planta induced necrosis. Site-directed mutagenesis on aromatic amino acid residues located within the CBDs as well as leaf infiltration assays using mutated and truncated recombinant proteins confirmed the importance of intact CBDs to induce defense responses. Tobacco and Arabidopsis thaliana leaf infiltration assays using synthetic peptides showed that the CBDs of CBEL are essential and sufficient to stimulate defense responses. Moreover, CBEL elicits a transient variation of cytosolic calcium levels in tobacco cells but not in protoplasts. These results define CBDs as a novel class of molecular patterns in oomycetes that are targeted by the innate immune system of plants and might act through interaction with the cell wall.