Caspase-Dependent Inhibition of Mousepox Replication by gzmB

Background

Ectromelia virus is a natural mouse pathogen, causing mousepox. The cytotoxic T (Tc) cell granule serine-protease, granzyme B, is important for its control, but the underlying mechanism is unknown. Using ex vivo virus immune Tc cells, we have previously shown that granzyme B is able to activate several independent pro-apoptotic pathways, including those mediated by Bid/Bak/Bax and caspases-3/-7, in target cells pulsed with Tc cell determinants.

Methods and Findings

Here we analysed the physiological relevance of those pro-apoptotic pathways in ectromelia infection, by incubating ectromelia-immune ex vivo Tc cells from granzyme A deficient (GzmB⁺ Tc cells) or granzyme A and granzyme B deficient (GzmA×B^−/− Tc cell) mice with ectromelia-infected target cells. We found that gzmB-induced apoptosis was totally blocked in ectromelia infected or peptide pulsed cells lacking caspases-3/-7. However ectromelia inhibited only partially apoptosis in cells deficient for Bid/Bak/Bax and not at all when both pathways were operative suggesting that the virus is able to interfere with apoptosis induced by gzmB in case not all pathways are activated. Importantly, inhibition of viral replication in vitro, as seen with wild type cells, was not affected by the lack of Bid/Bak/Bax but was significantly reduced in caspase-3/-7-deficient cells. Both caspase dependent processes were strictly dependent on gzmB, since Tc cells, lacking both gzms, neither induced apoptosis nor reduced viral titers.

Significance

Out findings present the first evidence on the biological importance of the independent gzmB-inducible pro-apoptotic pathways in a physiological relevant virus infection model.     

Discovery of a potent and selective aurora kinase inhibitor

This communication describes the discovery of a novel series of Aurora kinase inhibitors. Key SAR and critical binding elements are discussed. Some of the more advanced analogues potently inhibit cellular proliferation and induce phenotypes consistent with Aurora kinase inhibition. In particular, compound 21 (SNS-314) is a potent and selective Aurora kinase inhibitor that exhibits significant activity in pre-clinical in vivo tumor models.     

Cognitive behavioral therapy and fluoxetine for binge eating disorder: two-year follow-up

Objective: This study assessed the long‐term effects of group behavioral treatment plus individual cognitive behavioral therapy (CBT) and/or fluoxetine in binge eating disorder (BED) patients. Research Methods and Procedures: A total of 116 individuals were randomized to an initial five‐month trial and were followed up over two years. Assessments, including binge frequency, weight, and self‐report measures, were administered at pre‐treatment, post‐treatment, and ～6, 12, 18, and 24 months after initial treatment. Results: Across treatment groups, there was overall improvement over 29 months in binge frequency and in binge abstinence. The odds of binge abstinence 2 years post‐treatment were 1.373 times the odds of binge abstinence immediately post‐treatment. There was no significant change in weight over the two‐year period. Subjects who received individual CBT evidenced lower binge frequency over the two‐year follow‐up period than patients who had not received individual CBT. Similarly, CBT was associated with increased rates of binge abstinence. There were no main effects of treatment assignment on weight over the two‐year follow‐up period. There was a significant advantage for fluoxetine assignment over the two‐year follow‐up period on depressive symptoms. Discussion: The major significance of the study rests in its examination of the long‐term effects of standardized interventions for BED. Our findings provide support for the ideas that short‐term treatment may confer long‐term benefit and that not all treatments are equivalent in the benefits they confer.     

Sexually dimorphic SCAMP1 expression in the forebrain motor pathway for song production of juvenile zebra finches

Mechanisms regulating sexual differentiation of the zebra finch song system are not well understood. The present study was designed to more fully characterize secretory carrier membrane protein 1 (SCAMP1), which was identified in a cDNA microarray screen as showing increased expression in the forebrains of developing male compared with female zebra finches. We completed the sequence of the open reading frame and used in situ hybridization to compare mRNA in song control regions of juvenile (25-day-old) individuals. Expression was significantly greater in the HVC (used as a proper name) and robust nucleus of the arcopallium (RA) in males than in females. Immunohistochemistry revealed that SCAMP1 protein is also expressed in these two brain regions, and qualitatively appears greater in males. Western analysis confirmed that the protein is increased in the telencephalon of males when compared with females at 25 days of age. These results are consistent with the idea that SCAMP1 is involved in masculinization of these brain areas, perhaps facilitating the survival of cells within them.     

Acorn dispersal estimated by radio-tracking

Bird-dispersed seeds are difficult to track, especially in the case of long-distance dispersal events. To estimate the oak dispersal distance and the seed shadow generated by the European jay (Garrulus glandarius), we inserted radio-transmitters in 239 acorns, placed them in bird-feeders and then located them by radio-tracking. Using this methodology we located the exact caching site of 94 Quercus ilex and 54 Q. suber acorns and determined the caching habitat characteristics (vegetation type, distance, spatial distribution). The results show that: (1) there is no differences in the dispersal distance distribution between the different acorn species or sizes, (2) dispersal distances range from approximately 3 m up to approximately 550 m (mean = 68.6 m; median = 49.2 m), (3) recently abandoned fields and forest tracks were the sites preferred by jays to cache acorns, whereas fields and shrublands were avoided and (4) seed shadows showed acorn aggregation zones (i.e. clusters of caches) close to the feeder as well as isolated caches at longer distances. The results also suggest that radio-transmitters are a cheap and reliable way to determine seed shadows and quantify both seed dispersal and post-dispersal seed predation for medium to large seeds.     

Harvest prediction in ‘Algerie’ loquat

Plant phenology is in great measure driven by air temperature. To forecast harvest time for ‘Algerie’ loquat accurately, the growing degree days (GDD) needed from bloom to ripening were determined using data from nine seasons. The methods proposed by Zalom et al. (Zalom FG, Goodell PB, Wilson LT, Barnett WW, Bentley W, Degree-days: the calculation and use of heat units in pest management, leaflet no 21373, Division Agriculture and Natural Resources, University of California 10 pp, 1983) were compared as regards their ability to estimate heat summation based on hourly records. All the methods gave remarkably similar results for our cultivation area, although the double-sine method showed higher performance when temperatures were low. A base temperature of 3°C is proposed for ‘Algerie’ loquat because it provides a coefficient of variation in GDD among seasons of below 5%, and because of its compatibility with loquat growth. Based on these determinations, ‘Algerie’ loquat requires 1,715 GDD from bloom to harvest; under our conditions this heat is accumulated over an average of 159 days. Our procedure permits the ‘Algerie’ harvest date to be estimated with a mean error of 4.4 days (<3% for the bloom-harvest period). GDD summation did not prove superior to the use of the number of calendar days for predicting ‘Algerie’ harvest under non-limiting growing conditions. However, GDD reflects the developmental rate in water-stressed trees better than calendar days. Trees under deficit irrigation during flower development required more time and more heat to ripen their fruits.     

Pangnirtung Inuit and the Greenland Shark: Co-producing Knowledge of a Little Discussed Species

When faced with a species that is seldom encountered or discussed, can local or indigenous people piece together their accumulated experience to make inferences about the ecology of that species? In this paper the Greenland shark acts as a model to study how the Inuit of southern Baffin Island are able to produce ecological knowledge. We examine experiential information, reflections, variations in knowledge, and sense-making related to the Greenland Shark, and present a knowledge co-production process based on heuristic reasoning. The process of knowledge co-production has similarities to fuzzy logic, and highlights the adaptability and versatility of indigenous knowledge systems to generate new understandings about the species and its role in the Arctic marine environment. Interactions between the Inuit and researchers can provide a forum to facilitate knowledge co-production, and can be used as a strategy to engage the Indigenous and traditional peoples in resource management and conservation.     

Liver Stiffness Using Transient Elastography is Applicable to Canines for Hepatic Disease Models

Background

The objectives of this study were to evaluate the best position and best exploration probe for determining liver stiffness (LS) in dogs using transient liver elastography (TE). Thirteen dogs were used in the study.

Methodology/Principal Findings

Morphometric measurements taken were thoracic circumference, weight and height. Elastographic measurements were taken in 4 anatomical positions using two different probes: medium (M) and small (S). The exploration was considered correct when the success rate was above 60% and the interquartile range of the measurements did not exceed 30%. The best measurements were obtained in the middle of the 6th–9th intercostal spaces, with the dog in the left lateral position and using probe M for preference in adults and probe S mandatory for animals <2 years. The correlation between probes was 99%. Intra-observer variability showed an intra-class correlation of 97.6%.

Conclusions/Significance

TE is a technique that is reproducible in dogs.     

Inhibition of Angiotensin-Converting Enzyme Activity by Flavonoids: Structure-Activity Relationship Studies

Previous studies have demonstrated that certain flavonoids can have an inhibitory effect on angiotensin-converting enzyme (ACE) activity, which plays a key role in the regulation of arterial blood pressure. In the present study, 17 flavonoids belonging to five structural subtypes were evaluated in vitro for their ability to inhibit ACE in order to establish the structural basis of their bioactivity. The ACE inhibitory (ACEI) activity of these 17 flavonoids was determined by fluorimetric method at two concentrations (500 µM and 100 µM). Their inhibitory potencies ranged from 17 to 95% at 500 µM and from 0 to 57% at 100 µM. In both cases, the highest ACEI activity was obtained for luteolin. Following the determination of ACEI activity, the flavonoids with higher ACEI activity (i.e., ACEI >60% at 500 µM) were selected for further IC₅₀ determination. The IC₅₀ values for luteolin, quercetin, rutin, kaempferol, rhoifolin and apigenin K were 23, 43, 64, 178, 183 and 196 µM, respectively. Our results suggest that flavonoids are an excellent source of functional antihypertensive products. Furthermore, our structure-activity relationship studies show that the combination of sub-structures on the flavonoid skeleton that increase ACEI activity is made up of the following elements: (a) the catechol group in the B-ring, (b) the double bond between C2 and C3 at the C-ring, and (c) the cetone group in C4 at the C-ring. Protein-ligand docking studies are used to understand the molecular basis for these results.