Scission of COPI and COPII Vesicles Is Independent of GTP Hydrolysis

Intracellular transport and maintenance of the endomembrane system in eukaryotes depends on formation and fusion of vesicular carriers. A seeming discrepancy exists in the literature about the basic mechanism in the scission of transport vesicles that depend on GTP‐binding proteins. Some reports describe that the scission of COP‐coated vesicles is dependent on GTP hydrolysis, whereas others found that GTP hydrolysis is not required. In order to investigate this pivotal mechanism in vesicle formation, we analyzed formation of COPI‐ and COPII‐coated vesicles utilizing semi‐intact cells. The small GTPases Sar1 and Arf1 together with their corresponding coat proteins, the Sec23/24 and Sec13/31 complexes for COPII and coatomer for COPI vesicles were required and sufficient to drive vesicle formation. Both types of vesicles were efficiently generated when GTP hydrolysis was blocked either by utilizing the poorly hydrolyzable GTP analogs GTPγS and GMP‐PNP, or with constitutively active mutants of the small GTPases. Thus, GTP hydrolysis is not required for the formation and release of COP vesicles.     

DNA repair and free radicals, new insights into the mechanism of spore photoproduct lyase revealed by single amino acid substitution

The major DNA photoproduct in UV-irradiated Bacillus subtilis spores is the thymine dimer named spore photoproduct (SP, 5-(alpha-thyminyl)-5,6-dihydrothymine). The SP lesion has been found to be efficiently repaired by SP lyase (SPL) a very specific enzyme that reverses the SP to two intact thymines, at the origin of the great resistance of the spores to UV irradiation. SPL belongs to a superfamily of [4Fe-4S] iron-sulfur enzymes, called "Radical-SAM." Here, we show that the single substitution of cysteine 141 into alanine, a residue fully conserved in Bacillus species and previously shown to be essential for spore DNA repair in vivo, has a major impact on the outcome of the SPL-dependent repair reaction in vitro. Indeed the modified enzyme catalyzes the almost quantitative conversion of the SP lesion into one thymine and one thymine sulfinic acid derivative. This compound results from the trapping of the allyl-type radical intermediate by dithionite, used as reducing agent in the reaction mixture. Implications of the data reported here regarding the repair mechanism and the role of Cys-141 are discussed.     

Visual perception skills: a comparison between patients with Noonan syndrome and 22q11.2 deletion syndrome

Ventral and dorsal streams are visual pathways deputed to transmit information from the photoreceptors of the retina to the lateral geniculate nucleus and then to the primary visual cortex (V1). Several studies investigated whether one pathway is more vulnerable than the other during development, and whether these streams develop at different rates. The results are still discordant. The aim of the present study was to understand the functionality of the dorsal and the ventral streams in two populations affected by different genetic disorders, Noonan syndrome (NS) and 22q11.2 deletion syndrome (22q11.2DS), and explore the possible genotype–phenotype relationships. ‘Form coherence’ abilities for the ventral stream and ‘motion coherence’ abilities for the dorsal stream were evaluated in 19 participants with NS and 20 participants with 22q11.2DS. Collected data were compared with 55 age‐matched controls. Participants with NS and 22q11.2DS did not differ in the form coherence task, and their performance was significantly lower than that of controls. However, in the motion coherence task, the group with NS and controls did not differ, and both obtained significantly higher scores than the group with 22q11.2DS. Our findings indicate that deficits in the dorsal stream are related to the specific genotype, and that in our syndromic groups the ventral stream is more vulnerable than the dorsal stream.     

Effects of a Longitudinal Training Program on Responses to Exercise in Overweight Men

Objective: The aim of this study was to determine how training modifies metabolic responses and lipid oxidation in overweight young male subjects. Research Methods and Procedures: Eleven overweight subjects were selected for a 4‐month endurance training program. Before and after the training period, they cycled for 60 minutes at 50% of their Vo ₂max after an overnight fast or 3 hours after eating a standardized meal. Various metabolic and endocrine parameters, and respiratory exchange ratio values were evaluated. Results: Exercise‐induced plasma norepinephrine concentration increases were similar before and after training in fasted or fed conditions. After food intake, exercise promoted a decrease in plasma glucose and a higher increase in epinephrine than in fasting conditions. The increase in epinephrine after the meal was more marked after training (264 ± 32 vs. 195 ± 35 pg/mL). Training lowered the resting plasma nonesterified fatty acids. During exercise, changes in glycerol were similar to those found before training. Lipid oxidation during exercise was higher in fasting than in fed conditions (15.5 ± 1.4 vs. 22.3 ± 1.7 g/h). Training did not significantly increase fat oxidation when exercise was performed in fed conditions, but it did in fasting conditions (18.6 ± 1.4 vs. 27.2 ± 1.8 g/h). Discussion: Endurance training decreased plasma nonesterified fatty acids, cholesterol, and insulin concentrations. Training increased lipid oxidation during exercise, in fasting conditions, and not when exercise was performed after the meal. During exercise in overweight subjects, the fasting condition seems more suited to oxidizing fat and maintaining glucose homeostasis than a 3‐hour wait after a standard meal.     

Pest pressure,hurricanes, and genotype interact to strongly impact stem form in young loblolly pine (Pinus taeda L.) along the coastal plain of North Carolina

Key message

Stem defects in loblolly pine due to insect pests and wind damage can decrease economic value and affect ecosystem function, however silvicultural management can decrease the impact of these stresses.

Abstract

Loblolly pine (Pinus taeda L.) is one of the most important tree crops in the southern United States, comprising 45 % of commercial forestry land. Stem defects can reduce timber product quality and influence competitive interactions. We examined the effects of controlling Nantucket pine tip moth (Rhyacionia frustrata Scudder in Comstock, 1880) and site management (fertilizer or herbicide use) on stem defects, of two full-sib families (C1 and C2) and two clonal varieties (V1 and V2) of loblolly pine at upper and lower coastal plain sites in North Carolina (UCP and LCP, respectively). At UCP, V1 and V2 had fewer stem defects with insecticide treatment indicating that pest pressure affected stem form. C2 had twice as many defects as other genotypes at LCP, while C1 had the most defects at UCP, showing that carefully matching site and genotype could improve plantation performance without increasing costs. Additionally, we examined the effects of Hurricane Irene on stem leaning at LCP. V1, the tallest genotype, was most strongly affected, indicating genotype differences in tolerance to this form of abiotic stress. Interestingly, insecticide treatment decreased the negative effects of the hurricane on C1 stem lean, indicating that tip moth pressure may make C1 more susceptible. Our results illustrate that the interaction of biotic and abiotic stressors, such as pest infestation and climate, can strongly impact stem form, potentially affecting ecophysiological function and economic value. Cost-effective silvicultural options, such as pest control and management of genetic resources can potentially decrease exposure to such environmental risk.     

External validation of Adjuvant! Online breast cancer prognosis tool. Prioritising recommendations for improvement

Background

Adjuvant! Online is a web-based application designed to provide 10 years survival probability of patients with breast cancer. Several predictors have not been assessed in the original Adjuvant! Online study. We provide the validation of Adjuvant! Online algorithm on two breast cancer datasets, and we determined whether the accuracy of Adjuvant! Online is improved with other well-known prognostic factors.

Patients and Methods

The French data set is composed of 456 women with early breast cancer. The Dutch data set is composed of 295 women less than 52 years of age. Agreement between observation and Adjuvant! Online prediction was checked, and logistic models were performed to estimate the prognostic information added by risk factors to Adjuvant! Online prediction.

Results

Adjuvant! Online prediction was overall well-calibrated in the French data set but failed in some subgroups of such high grade and HER2 positive patients. HER2 status, Mitotic Index and Ki67 added significant information to Adjuvant! Online prediction. In the Dutch data set, the overall 10-year survival was overestimated by Adjuvant! Online, particularly in patients less than 40 years old.

Conclusion

Adjuvant! Online needs to be updated to adjust overoptimistic results in young and high grade patients, and should consider new predictors such as Ki67, HER2 and Mitotic Index.     

Ajuba: a new microtubule-associated protein that interacts with BUBR1 and Aurora B at kinetochores in metaphase

Background information. The role of the LIM‐domain‐containing protein Ajuba was initially described in cell adhesion and migration processes and recently in mitosis as an activator of the Aurora A kinase. Results. In the present study, we show that Ajuba localizes to centrosomes and kinetochores during mitosis. This localization is microtubule‐dependent and Ajuba binds microtubules in vitro. A microtubule regrowth assay showed that Ajuba follows nascent microtubules from centrosomes to kinetochores. Owing to its contribution to mitotic commitment and its microtubule‐dependent localization, Ajuba could also play a role during the metaphase—anaphase transition. We show that Ajuba interacts with Aurora B and BUBR1 [BUB (budding uninhibited by benomyl)‐related 1], two major components of the mitotic checkpoint. Inhibition of BUBR1 by siRNA (small interfering RNA) disrupts chromosome alignment at the metaphase plate and modifies Ajuba localization due to premature mitotic exit. Conclusions. Ajuba is a microtubule‐associated protein that collaborates with Aurora B and BUBR1 at the metaphase—anaphase transition and this may be important to ensure proper chromosome segregation.     

Tracing hotspots of soil erosion in high mountain environments: how forensic science based on plant eDNA can lead the way. An opinion

Plant and Soil - High mountain environments are among the most fragile on Earth. Due to anthropogenic disturbances and the exposure to extreme weather events, the rates of soil erosion have...     

Shear Viscosity Investigation on Mango Juice with High Frequency Longitudinal Ultrasonic Waves and Rotational Viscosimetry

Ultrasonic velocity and attenuation measurements were performed on mango juices at 25 MHz in order to estimate longitudinal viscosity. Juices were extracted from fruits, removed periodically from fruit batches undergoing ripening for 3 weeks under controlled conditions. The correlation between longitudinal viscosity and apparent dynamic shear viscosity, obtained from flow tests, showed that up to 12–13 wt.% of Soluble Solids Content (SSC), the juices presented a Newtonian behavior. In this case the relation between longitudinal viscosity measured by ultrasound and shear viscosity measured by flow tests was very simple leading to the conclusion that ultrasound could replace rotating viscosimeters for specific applications. Over this limit, the results were also clearly correlated but the correlation depended on the shear rate because of the shear thinning behavior of the juices certainly due to soluble pectins. The use of longitudinal ultrasonic waves as a tool for viscosity determination on large batches of samples is discussed at the end of this communication.