Autoinhibition Mechanism of the Ubiquitin-Conjugating Enzyme UBE2S by Autoubiquitination

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The Diversity and Evolution of Wolbachia Ankyrin Repeat Domain Genes

Ankyrin repeat domain-encoding genes are common in the eukaryotic and viral domains of life, but they are rare in bacteria, the exception being a few obligate or facultative intracellular Proteobacteria species. Despite having a reduced genome, the arthropod strains of the alphaproteobacterium Wolbachia contain an unusually high number of ankyrin repeat domain-encoding genes ranging from 23 in wMel to 60 in wPip strain. This group of genes has attracted considerable attention for their astonishing large number as well as for the fact that ankyrin proteins are known to participate in protein-protein interactions, suggesting that they play a critical role in the molecular mechanism that determines host-Wolbachia symbiotic interactions. We present a comparative evolutionary analysis of the wMel-related ankyrin repeat domain-encoding genes present in different Drosophila-Wolbachia associations. Our results show that the ankyrin repeat domain-encoding genes change in size by expansion and contraction mediated by short directly repeated sequences. We provide examples of intra-genic recombination events and show that these genes are likely to be horizontally transferred between strains with the aid of bacteriophages. These results confirm previous findings that the Wolbachia genomes are evolutionary mosaics and illustrate the potential that these bacteria have to generate diversity in proteins potentially involved in the symbiotic interactions.     

Respiratory distress syndrome (RDS) in premature infants is underscored by the magnitude of Th1 cytokine polarization

Respiratory distress syndrome (RDS) is a common problem and the leading cause of death in premature infants (PI). The introduction of surfactant treatment for RDS management has lowered mortality and morbidity; nevertheless, some neonates do not improve and are at increased risk of pulmonary hemorrhage. Inflammation, not only local but also systemic, seems to play an important role in the pathogenesis of RDS. To determine whether cytokine patterns characterize RDS and its outcome, we measured type-1 (IL-2, TNF-α, IFN-γ, IL-6) and type-2 (IL-4, IL-5, IL-10, TGF-β1) serum cytokines of 47 PI with established RDS and a control group of 30 healthy, appropriate for gestational age, full-term neonates. Cord blood samples were obtained at the time of delivery from PI and controls. Venous blood samples were collected from PI who received surfactant treatment and/or developed pulmonary hemorrhage. Significantly elevated cord blood cytokine levels were observed in PI at time of delivery, compared to controls, except for IL-5 and TNF-α levels that were within control range. The type-1/type-2 cytokine ratio was significantly increased in PI vs controls. Neonates who developed pulmonary hemorrhage between 2 and 3 days of life and/or died, presented the strongest Th1 and type-1 cytokine polarization that was mainly due to increased IFN-γ and TNF-α, and decreased TGF-β1. The majority of these PI were female with very low gestational age. Overall, PI with RDS present a Th1/type-1 cytokine polarization, which persists irrespective of the treatment provided, and is amplified when complications appear. Th1 polarization is associated with poor prognosis.     

Little effect of delayed mating on fecundity or fertility of female fungus gnats Lycoriella ingenua

The effect of delayed female mating for the mushroom fungus gnat Lycoriella ingenua is investigated. We examine the effect of delaying female mating on the fertility and egg viability of female flies that have a mating delay of 0–5 days after emergence. Male fly age is held constant. Female age does not impact male acceptance and most flies copulate within seconds of pairing. We find that female flies experiencing mating delays of 0–4 days after emergence lay a similar number of eggs onto artificial substrates. Females that experience a mating delay of 5 days lay 54% fewer eggs than those that mate on day 0 (day of emergence). There is no effect of mating delay on the percentage of larvae that emerge. The results of the present study indicate that mating delays have little effect on the fertility or fecundity of the mushroom fungus pest L. ingenua.     

Cerebral metabolism during upper and lower body exercise.

When continuation of exercise calls for a "will," the cerebral metabolic ratio of O2 to (glucose + lactate) decreases, with the largest reduction (30-50%) at exhaustion. Because a larger effort is required to exercise with the arms than with the legs, we tested the hypothesis that the reduction in the cerebral metabolic ratio would become more pronounced during arm cranking than during leg exercise. The cerebral arterial-venous differences for blood-gas variables, glucose, and lactate were evaluated in two groups of eight subjects during exhaustive arm cranking and leg exercise. During leg exercise, exhaustion was elicited after 25 +/- 6 (SE) min, and the cerebral metabolic ratio was reduced from 5.6 +/- 0.2 to 3.5 +/- 0.2 after 10 min and to 3.3 +/- 0.3 at exhaustion (P < 0.05). Arm cranking lasted for 35 +/- 4 min and likewise decreased the cerebral metabolic ratio after 10 min (from 6.7 +/- 0.4 to 5.0 +/- 0.3), but the nadir at exhaustion was only 4.7 +/- 0.4, i.e., higher than during leg exercise (P < 0.05). The results demonstrate that exercise decreases the cerebral metabolic ratio when a conscious effort is required, irrespective of the muscle groups engaged. However, the comparatively small reduction in the cerebral metabolic ratio during arm cranking suggests that it is influenced by the exercise paradigm.     

The side-effects of plant protection products used in olive cultivation on the hymenopterous egg parasitoid Trichogramma cacoeciae Marchal

Abstract:  Six insecticides (Malathion, Quik, Cidial, Dimethoate, Actellic, Deltamethrin) and two mineral oils (Super misrona and Kemesol) are currently used to control the most important pests in Mediterranean olive cultivation (olive fly, olive moth and black scale). Potential side-effects of these compounds were tested on the egg parasitoid Trichogramma cacoeciae Marchal, following recommendations of the working group 'Pesticides and Beneficial Organisms' of the International Organization for Biological Control, West Palaearctic Regional Section (IOBC/WPRS). In the present study, three different types of test methods were carried out: (1) initial toxicity dose–response test on adult wasps; (2) initial toxicity test on pupae, using field recommended rates; and (3) persistent test on olive foliage to assess the duration of harmful activity. The six insecticides tested reduced parasitism by 80–95% and rated as moderately harmful at the field recommended doses. The two mineral oils reduced parasitism up to 25% and were therefore rated as harmless to the adult stage of the parasitoid. The results of the pupal test (parasitoid pupa inside the host egg) showed that Malathion was harmless; Quik, Actellic and Cidial were slightly harmful, while Dimethoate and Deltamethrin were moderately harmful. The results of the persistence test on olive foliage showed that Malathion, Quik and Actellic were slightly persistent, while Cidial, Dimethoate and Deltamethrin were moderately persistent.     

CFTR gene mutations – including three novel nucleotide substitutions – and haplotype background in patients with asthma, disseminated bronchiectasis and chronic obstructive pulmonary disease

In order to investigate the incidence of cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations and unclassified variants in chronic pulmonary disease in children and adults, we studied 20 patients with asthma, 19 with disseminated bronchiectasis (DB) of unknown aetiology, and 12 patients with chronic obstructive pulmonary disease (COPD), and compared the results to 52 subjects from the general Greek population. Analysis of the whole coding region of the CFTR gene and its flanking intronic regions revealed that the proportion of CFTR mutations was 45% in asthma (P<0.05), 26.3% in DB (P>0.05), 16.7% in COPD (P>0.05), compared to 15.4% in the general population. Seventeen different molecular defects involved in disease predisposition were identified in 16 patients. Three potentially disease-causing mutations, T388 M, M1R and V11I, are novel, found so far only in three asthma patients. The hyperactive M470 allele was found more frequently in COPD patients (frequency 70.8%, P<0.01) than in the controls. The study of the TGmTnM470 V polyvariant CFTR allele revealed the presence of CFTR function-modulating haplotypes TG13/T5/M470, TG11/T5/M470, TG12/T5/V470 and TG12/T7, combined with M470 or V470, in six asthma patients, four DB patients (P<0.01), and two COPD patients (P<0.05). These results confirm the involvement of the CFTR gene in asthma, DB and possibly in COPD.     

Correlation of the Insecticidal Activity of the Bacillus thuringiensis A4 Strain Against Bactrocera oleae (Diptera) with the 140-kDa Crystal Polypeptide

The crystals of the soil-isolated Bacillus thuringiensis (Bt) strain A4 consist of two polypeptides with molecular mass of 140 kDa and 32 kDa that exhibit insecticidal activity against adult flies of Bactrocera oleae (Diptera). Plasmid curing applied to this strain resulted in the isolation of several subclones exhibiting alterations in their crystal polypeptides as well as two acrystalliferous subclones. The crystals of subclone 1.1 lacked the 32-kDa polypeptide and consisted uniquely of a 140-kDa polypeptide antigenically related to the parental 140-kDa crystal polypeptide. Additionally, the crystals of this subclone exhibited insecticidal activity against B. oleae equivalent to that of the parental strain. Therefore, the 32-kDa crystal polypeptide is dispensable for insecticidal activity, which appears to be dependent on the presence of the 140-kDa crystal polypeptide. Received: 5 April 2000 / Accepted 2 May 2000     

Wolf depredation on livestock in central Greece

We studied wolfCanis lupus Linnaeus, 1758 — livestock conflict in central Greece by investigating patterns of 267 verified wolf attacks on livestock for 21 months. Wolves attacked adult goats 43% and cattle 218% more than expected, whereas sheep 41% less than expected from their availability. Wolves killed less than four sheep or goats in 79%, and one cow or calf in 74% of depredation events, respectively. We recorded higher attack rates during wolf post-weaning season. Wolf attacks on strayed, or kept inside non predator-proof enclosures, sheep and goats, were on average two to four times respectively more destructive than those when livestock was guarded by a shepherd. Sheepdog use reduced losses per attack. Optimal sheepdog number ranged from 3 to 9 animals depending on flock size. Losses per attack were positively related to the number of wolves involved. Total losses per farm were positively correlated with the size of livestock unit but percentage losses per capita increased with decreasing flock size. Management implications to mitigate livestock depredation are discussed.