A rapid and sensitive ELISA to quantify an HBsAg specific monoclonal antibody and a plant-derived antibody during their downstream purification process.

An enzyme-linked immunosorbent assay to quantify the mAb CB.Hep-1 during downstream purification process was standardized and validated. This assay is characterized by a short time of incubation at high temperature, allowing the detection of this antibody with high specificity and sensitivity. Detection of antigen-antibody reaction was achieved using a horseradish peroxidase conjugated anti-mouse IgG whose enzyme activity was revealed with o-phenylenediamine substrate. The immunoassay is linear in a range between 3.12 and 50 ng/mL, with a recovery of 98.55-107.62%. According to results, it is possible to estimate the mAb CB.Hep-1 concentration with high precision and reproducibility. The intra- and interassay coefficient of variation ranged from 0.25 to 8.64% and 1.84 to 9.43%, respectively. Significant differences were not observed in the plant-derived antibody quantification by HRP-ELISA and PhoA-ELISA (n=18), demonstrating that plant endogenous peroxidases do not produce interferences in the quantification of this molecule. Therefore, both antibodies can be tested with the same immunoassay with high precision, specificity and accuracy during their respective purification processes without interference of the buffers and sample characteristics.     

An improved radiological method for the evaluation of Achilles tendon xanthomatosis

Xanthomatous infiltration of tendons is a clinical feature common to many cases of hyperlipidemia. The xanthomas can be detected and only grossly assessed by palpation. This report describes a radiological technique used to assess these lesions at the Achilles tendon level. The “mammography technique” applied to the study of Achilles tendons was used in 32 normolipemic subjects and 32 hyperlipidemic patients. Both tendons could be observed in their entire length and their thickness, greatly increased in xanthomatosis, could be accurately measured. The results of this survey suggest that the radiological approach may provide a useful tool for the routine evaluation and follow-up of tendon xanthomas.     

Ecological and socio-economic factors affecting extinction risk in parrots

Parrots (Psittaciformes) are among the most threatened bird orders with 28 % (111 of 398) of extant species classified as threatened under IUCN criteria. We confirmed that parrots have a lower Red List Index (higher aggregate extinction risk) than other comparable bird groups, and modeled the factors associated with extinction risk. Our analyses included intrinsic biological, life history and ecological attributes, external anthropogenic threats, and socio-economic variables associated with the countries where the parrot species occur, while we controlled for phylogenetic dependence among species. We found that the likelihood of parrot species being classified as threatened was less for species with larger historical distribution size, but was greater for species with high forest dependency, large body size, long generation time, and greater proportion of the human population living in urban areas in the countries encompassing the parrots’ home ranges. The severity of extinction risk (from vulnerable to critically endangered) was positively related to the per capita gross domestic product (GDP) of the countries of occurrence, endemism to a single country, and lower for species used as pets. A disproportionate number of 16 extinct parrot species were endemic to islands and single countries, and were large bodied, habitat specialists. Agriculture, hunting, trapping, and logging are the most frequent threats to parrots worldwide, with variation in importance among regions. We use multiple methods to rank countries with disproportionately high numbers of threatened parrot species. Our results promote understanding of global and regional factors associated with endangerment in this highly threatened taxonomic group, and will enhance the prioritization of conservation actions.     

ASG2 is a farnesylated DWD protein that acts as ABA negative regulator in Arabidopsis

The tagging‐via‐substrate approach designed for the capture of mammal prenylated proteins was adapted to Arabidopsis cell culture. In this way, proteins are in vivo tagged with an azide‐modified farnesyl moiety and captured thanks to biotin alkyne Click‐iT® chemistry with further streptavidin‐affinity chromatography. Mass spectrometry analyses identified four small GTPases and ASG2 (ALTERED SEED GERMINATION 2), a protein previously associated to the seed germination gene network. ASG2 is a conserved protein in plants and displays a unique feature that associates WD40 domains and tetratricopeptide repeats. Additionally, we show that ASG2 has a C‐terminal CaaX‐box that is farnesylated in vitro. Protoplast transfections using CaaX prenyltransferase mutants show that farnesylation provokes ASG2 nucleus exclusion. Moreover, ASG2 interacts with DDB1 (DAMAGE DNA BINDING protein 1), and the subcellular localization of this complex depends on ASG2 farnesylation status. Finally, germination and root elongation experiments reveal that asg2 and the farnesyltransferase mutant era1 (ENHANCED RESPONSE TO ABSCISIC ACID (ABA) 1) behave in similar manners when exposed to ABA or salt stress. To our knowledge, ASG2 is the first farnesylated DWD (DDB1 binding WD40) protein related to ABA response in Arabidopsis that may be linked to era1 phenotypes.     

Structure-activity relationships of 4-hydroxy-4-biaryl-proline acylsulfonamide tripeptides: A series of potent NS3 protease inhibitors for the treatment of hepatitis C virus

The design and synthesis of a series of tripeptide acylsulfonamides as potent inhibitors of the HCV NS3/4A serine protease is described. These analogues house a C4 aryl, C4 hydroxy-proline at the S2 position of the tripeptide scaffold. Information relating to structure-activity relationships as well as the pharmacokinetic and cardiovascular profiles of these analogues is provided.     

Distinct sources of gene flow produce contrasting population genetic dynamics at different range boundaries of a Choristoneura budworm

Populations are often exposed to multiple sources of gene flow, but accounts are lacking of the population genetic dynamics that result from these interactions or their effects on local evolution. Using a genomic clines framework applied to 1,195 single nucleotide polymorphisms, we documented genomewide, locus‐specific patterns of introgression between Choristoneura occidentalis biennis spruce budworms and two ecologically divergent relatives, C. o. occidentalis and Choristoneura fumiferana, that it interacts with at alternate boundaries of its range. We observe contrasting hybrid indexes between the two hybrid zones, no overlap in “gene‐flow outliers” (clines showing relatively extreme extents or rates of locus‐specific introgression) and variable linkage disequilibrium among those outliers. At the same time, correlated genomewide rates of introgression between zones suggest the presence of processes common to both boundaries. These findings highlight the contrasting population genetic dynamics that can occur at separate frontiers of a single population, while also suggesting that shared patterns may frequently accompany cases of divergence‐with‐gene‐flow that involve a lineage in common. Our results point to potentially complex evolutionary outcomes for populations experiencing multiple sources of gene flow.     

Pyrano-[2,3b]-pyridines as potassium channel antagonists

The design and synthesis of a series of highly functionalized pyrano-[2,3b]-pyridines is described. These compounds were assayed for their ability to block the I(Kur) channel encoded by the gene hKV1.5 in patch-clamped L-929 cells. Six of the compounds in this series showed sub-micromolar activity, the most potent being 4-(4-ethyl-benzenesulfonylamino)-3-hydroxy-2,2-dimethyl-3,4-dihydro-2H-pyrano[2,3b]-pyridine-6-carboxylic acid ethyl-phenyl-amide with an IC(50) of 378 nM.     

Increased liver glycogen levels enhance exercise capacity in mice

Muscle glycogen depletion has been proposed as one of the main causes of fatigue during exercise. However, few studies have addressed the contribution of liver glycogen to exercise performance. Using a low-intensity running protocol, here, we analyzed exercise capacity in mice overexpressing protein targeting to glycogen (PTG) specifically in the liver (PTG^OE mice), which show a high concentration of glycogen in this organ. PTG^OE mice showed improved exercise capacity, as determined by the distance covered and time ran in an extenuating endurance exercise, compared with control mice. Moreover, fasting decreased exercise capacity in control mice but not in PTG^OE mice. After exercise, liver glycogen stores were totally depleted in control mice, but PTG^OE mice maintained significant glycogen levels even in fasting conditions. In addition, PTG^OE mice displayed an increased hepatic energy state after exercise compared with control mice. Exercise caused a reduction in the blood glucose concentration in control mice that was less pronounced in PTG^OE mice. No changes were found in the levels of blood lactate, plasma free fatty acids, or β-hydroxybutyrate. Plasma glucagon was elevated after exercise in control mice, but not in PTG^OE mice. Exercise-induced changes in skeletal muscle were similar in both genotypes. These results identify hepatic glycogen as a key regulator of endurance capacity in mice, an effect that may be exerted through the maintenance of blood glucose levels.