Active Mechanics Reveal Molecular-Scale Force Kinetics in Living Oocytes

Active diffusion of intracellular components is emerging as an important process in cell biology. This process is mediated by complex assemblies of molecular motors and cytoskeletal filaments that drive force generation in the cytoplasm and facilitate enhanced motion. The kinetics of molecular motors have been precisely characterized in vitro by single molecule approaches, but their in vivo behavior remains elusive. Here, we study the active diffusion of vesicles in mouse oocytes, where this process plays a key role in nuclear positioning during development, and combine an experimental and theoretical framework to extract molecular-scale force kinetics (force, power stroke, and velocity) of the in vivo active process. Assuming a single dominant process, we find that the nonequilibrium activity induces rapid kicks of duration τ ～ 300 μs resulting in an average force of F ～ 0.4 pN on vesicles in in vivo oocytes, remarkably similar to the kinetics of in vitro myosin-V. Our results reveal that measuring in vivo active fluctuations allows extraction of the molecular-scale activity in agreement with single-molecule studies and demonstrates a mesoscopic framework to access force kinetics.     

Approaches to the study of mycorrhizas in Romania

Research on mycorrhizas in Romania started some 60 years ago, first in forestry, then in agriculture and finally in horticulture. The main studies have covered the following topics: (i) The mycorrhizal symbiosis; (ii) Ectomycorrhizas in natural forest ecosystems; (iii) Mycorrhizas in agriculture and forestry; (iv) Field methods, e.g., the development and strategies for obtaining high colonization rates in crops; interactions between mycorrhizal fungi, and soil microorganisms including Rhizobium spp.; factors influencing the efficiency of the mycorrhizas; (v) Management of mycorrhizas in agriculture, horticulture, and forestry as well as management of edible mycorrhizal mushrooms; (vi) Biological protection of crops against root pathogens (Pythium, Phytophthora, Fusarium etc.) using inoculation with mycorrhizal fungi; (vii) Architecture, anatomy and physiology of mycorrhizal plants; (viii) Isolation techniques; methods for investigating mycorrhizas; (ix) Identification by classical and modern methods and characterization of mycorrhizal fungi.     

Genome-Wide Prediction Methods in Highly Diverse and Heterozygous Species: Proof-of-Concept through Simulation in Grapevine

Nowadays, genome-wide association studies (GWAS) and genomic selection (GS) methods which use genome-wide marker data for phenotype prediction are of much potential interest in plant breeding. However, to our knowledge, no studies have been performed yet on the predictive ability of these methods for structured traits when using training populations with high levels of genetic diversity. Such an example of a highly heterozygous, perennial species is grapevine. The present study compares the accuracy of models based on GWAS or GS alone, or in combination, for predicting simple or complex traits, linked or not with population structure. In order to explore the relevance of these methods in this context, we performed simulations using approx 90,000 SNPs on a population of 3,000 individuals structured into three groups and corresponding to published diversity grapevine data. To estimate the parameters of the prediction models, we defined four training populations of 1,000 individuals, corresponding to these three groups and a core collection. Finally, to estimate the accuracy of the models, we also simulated four breeding populations of 200 individuals. Although prediction accuracy was low when breeding populations were too distant from the training populations, high accuracy levels were obtained using the sole core-collection as training population. The highest prediction accuracy was obtained (up to 0.9) using the combined GWAS-GS model. We thus recommend using the combined prediction model and a core-collection as training population for grapevine breeding or for other important economic crops with the same characteristics.     

The Adult Cystic Fibrosis Airway Microbiota Is Stable over Time and Infection Type,and Highly Resilient to Antibiotic Treatment of Exacerbations

Cystic fibrosis (CF) is characterized by defective mucociliary clearance and chronic airway infection by a complex microbiota. Infection, persistent inflammation and periodic episodes of acute pulmonary exacerbation contribute to an irreversible decline in CF lung function. While the factors leading to acute exacerbations are poorly understood, antibiotic treatment can temporarily resolve pulmonary symptoms and partially restore lung function. Previous studies indicated that exacerbations may be associated with changes in microbial densities and the acquisition of new microbial species. Given the complexity of the CF microbiota, we applied massively parallel pyrosequencing to identify changes in airway microbial community structure in 23 adult CF patients during acute pulmonary exacerbation, after antibiotic treatment and during periods of stable disease. Over 350,000 sequences were generated, representing nearly 170 distinct microbial taxa. Approximately 60% of sequences obtained were from the recognized CF pathogens Pseudomonas and Burkholderia, which were detected in largely non-overlapping patient subsets. In contrast, other taxa including Prevotella, Streptococcus, Rothia and Veillonella were abundant in nearly all patient samples. Although antibiotic treatment was associated with a small decrease in species richness, there was minimal change in overall microbial community structure. Furthermore, microbial community composition was highly similar in patients during an exacerbation and when clinically stable, suggesting that exacerbations may represent intrapulmonary spread of infection rather than a change in microbial community composition. Mouthwash samples, obtained from a subset of patients, showed a nearly identical distribution of taxa as expectorated sputum, indicating that aspiration may contribute to colonization of the lower airways. Finally, we observed a strong correlation between low species richness and poor lung function. Taken together, these results indicate that the adult CF lung microbiome is largely stable through periods of exacerbation and antibiotic treatment and that short-term compositional changes in the airway microbiota do not account for CF pulmonary exacerbations.     

Inactivation of a Novel Gene Produces a Phenotypic Variant Cell and Affects the Symbiotic Behavior of Xenorhabdus nematophilus

Xenorhabdus nematophilus is an insect pathogen that lives in a symbiotic association with a specific entomopathogenic nematode. During prolonged culturing, variant cells arise that are deficient in numerous properties. To understand the genetic mechanism underlying variant cell formation, a transposon mutagenesis approach was taken. Three phenotypically similar variant strains of X. nematophilus, each of which contained a single transposon insertion, were isolated. The insertions occurred at different locations in the chromosome. The variant strain, ANV2, was further characterized. It was deficient in several properties, including the ability to produce antibiotics and the stationary-phase-induced outer membrane protein, OpnB. Unlike wild-type cells, ANV2 produced lecithinase. The emergence of ANV2 from the nematode host was delayed relative to the emergence of the parental strain. The transposon in ANV2 had inserted in a gene designated var1, which encodes a novel protein composed of 121 amino acid residues. Complementation analysis confirmed that the pleiotropic phenotype of the ANV2 strain was produced by inactivation of var1. Other variant strains were not complemented by var1. These results indicate that inactivation of a single gene was sufficient to promote variant cell formation in X. nematophilus and that disruption of genetic loci other than var1 can result in the same pleiotropic phenotype.     

Effect of production system and pruning on Aphis sambuci dynamics over time and on elderberry yield

In a 2‐year study, elder aphid (Aphis sambuci) dynamics over time and berry yield were evaluated in two production systems (integrated and organic) and in two winter pruning treatments (trees pruned to four and eight scaffolds) in two black elderberry orchards in Hungary. In the organic production system, the first aphid colony was observed 1–2 weeks earlier (late‐March) in both years and locations compared to the integrated programme. The number of aphid colonies then increased until mid‐May in both years, reaching a maximum number of aphid colonies of 11.2 on 100 scaffolds in the integrated production system and of 38.9 in the organic programme. Then, the number of colonies decreased and reached a zero value at mid‐June in the integrated production system and 2 weeks later (early July) in the organic one in both years and locations. First autumn aphid colonies were observed in early September in the integrated production system but 2 weeks earlier (late August) in the organic one in both years and locations. The number of aphid colonies between mid‐April and mid‐June indicated a larger increase on trees pruned to eight scaffolds compared to trees pruned to four scaffolds. Both the total number of aphid colonies and the area under the aphid colony curves (AUACC) were significantly lower (P < 0.001) in the integrated treatments compared with organic ones. Across all treatments, both measures were significantly lower (P < 0.05) on trees pruned to four scaffolds compared with trees pruned to eight scaffolds. However, when the effect of pruning on the number of aphid colonies was analysed separately for integrated and organic plots, pruning caused significant differences in aphid colony numbers and AUACC in the organic plots. Berry yield was significantly higher (P < 0.05) in the integrated treatments compared with the organic ones, but pruning showed no significant effect on yield. Overall, pruning to four scaffolds resulted in a lower aphid colony number in the organic production system compared to the integrated one. Thus, winter pruning may be useful as an aphid control strategy in organic elderberry orchards.     

An epidemiologic study of hypertension in Newfoundland

The distribution of arterial blood pressure (BP) values of 1499 adult inhabitants of four Newfoundland communities was surveyed. Mean age- and sex-adjusted BP scores were found to be higher in each of three fishing villages than in the logging and mining community, Badger, in the central part of the province. It is postulated that the observed differences may be caused by environmental factors. The prevalence of individuals with probable essential hypertension (diastolic BP > 100 mm. Hg) in individuals over 50 years of age was found to be three times higher in the island community, Fogo, than in Badger. In both communities less than one third of those with probable hypertensive disease had received treatment.