Specificity of coenzyme Q10 for a balanced function of respiratory chain and endogenous ubiquinone biosynthesis in human cells

Coenzyme Q (Q) is an obligatory component of both respiratory chain and uncoupling proteins. Also, Q acts as an antioxidant in cellular membranes. Several neurodegenerative diseases are associated with modifications of Q10 levels. For these reasons, therapies based on Q supplementation in the diet are currently studied in order to mitigate the symptoms of these diseases. However, the incorporation of exogenous Q also affects aging process in nematodes probably affecting reactive oxygen species (ROS) production. The aim of the present work is to clarify if supplementation with both Q10 and Q6 isoforms affects mitochondrial Q10 content, respiratory chain activity and ROS levels in human cells. Cells incorporated exogenously added Q10 and Q6 isoforms into mitochondria that produced changes in mitochondrial activity depending on the side chain length. Supplementation with Q10, but not with Q6, increased mitochondrial Q-dependent activities. However, Q6 affected the mitochondrial membrane potential, ROS production, and increased the protein levels of both catalase and Mn-superoxide dismutase (Mn-SOD). Also, Q6 induced a transient decrease in endogenous mitochondrial Q10 levels by increasing its catabolism. These results show that human cells supplemented with Q6 undergo a mitochondrial impairment, which is not observed with Q10 supplementation.     

Trichomonas vaginalis: characterization of a 39-kDa cysteine proteinase found in patient vaginal secretions

Trichomonosis, a chronic sexually transmitted disease, remains a public health problem affecting yearly over 170 million people worldwide. This disease is caused by Trichomonas vaginalis, a protozoan flagellate rich in cysteine proteinases (CPs). Although CPs are involved in trichomonal cytopathogenicity, only few of them have been defined as virulence factors. In this study, we characterize a T. vaginalis 39-kDa proteinase (CP39) found in vaginal secretions from patients with trichomonosis. The CP39 proteinase bound to HeLa epithelial cells, vaginal epithelial cells (VECs), and human prostatic cancer cells (DU-145). CP39 did not bind to a human colon cancer (CaCo) cell line, suggesting tissue-specific binding. CP39 was found in six fresh trichomonad isolates tested. In two-dimensional gels, CP39 appeared as a single spot with a pI 4.5. CP39 is inhibited by E-64, stable at 50 degrees C, and active in a wide pH range (3.6-9.0), with an optimum pH at 7.0. In addition, CP39 degraded collagens I, III, IV, and V, human fibronectin, human hemoglobin, and human immunoglobulins A and G. Indirect immunofluorescence detected CP39 on the parasite surface with specific polyclonal antibody to purified CP39. Finally, CP39 was found to be immunogenic, as evidenced by detection on immunoblots with serum of patients with trichomonosis, but not control individuals. These data suggest that CP39 may play a role during trichomonal infection.     

Caveolae are a novel pathway for membrane-type 1 matrix metalloproteinase traffic in human endothelial cells

The extracellular matrix (ECM) distinctly modulates membrane type 1-matrix metalloproteinase (MT1-MMP) in human endothelial cells (ECs). Herein, ECM-dependent RhoA activation is shown to regulate MT1-MMP localization and activity as well as clathrin-independent internalization in confluent ECs. In this regard, caveolae are revealed as the major MT1-MMP endocytic pathway in human ECs. Thus, MT1-MMP is present at caveolae with caveolin-1 and both proteins together with alpha v beta 3 integrin colocalize at endothelial motility-associated extensions. Remarkably, caveolae traffic is required for proper MT1-MMP localization, activity, and function in migratory ECs as demonstrated by both treatment with caveolae-disrupting agents or selective targeting caveolin-1 expression by interference RNA. Thus, caveolae-mediated traffic constitutes a novel mechanism for MT1-MMP regulation in ECs during angiogenesis.     

Decoy presentations as a means to manipulate the risk of extrapair copulation: an experimental study in a semicolonial raptor, the Montagu's harrier (Circus pygargus)

Mate guarding and frequent copulations are two alternative paternity assurance strategies found in birds. In species with intensecourtship feeding, like raptors, the "frequent copulation"strategy is expected because male food provisioning conflictswith mate guarding. We evaluated experimentally the paternityassurance behavior of a semicolonial raptor, the Montagu'sharrier Circus pygargus, using decoy presentations to simulateterritorial intrusions. Breeding pairs were exposed to maleand female decoys at different periods during the female's reproductive cycle. Agonistic responses to decoys were intra-sexual,and the timing and intensity of male attacks toward male decoyssupported responses related to the risk of extrapair copulation(EPC): Male aggression peaked during the presumed fertile periodand almost disappeared after clutch completion. During thefertile period, copulation rate was significantly higher, andcopulations lasted longer, during male decoy presentations than during controls. Males also spent more time close to the femaleduring male decoy presentations compared to controls, bothduring the early prelaying and fertile periods, but not duringincubation. In the fertile period, males also increased presencetime close to the female in the hour following the removal of the male decoy. Conversely, female decoy presentations hadno significant effect on copulatory behavior or male presencetime. These results showed that the risk of EPC can be experimentallymanipulated by the means of decoy presentations, simulatingmale territorial intrusions, and that male Montagu's harriersincrease their short-term copulation frequency and female surveillancewhen they perceive themselves at an increased EPC risk.     

Lek mating systems: a case study in the Little Bustard Tetrax tetrax

Leks have recently been defined as male display aggregations that females attend primarily for the purpose of mating. This is an extended version of previous definitions, as a clear-cut definition of leks is difficult to obtain. Four criteria should be verified to identify a lekking species: (i) there is no male parental investment beyond the sperm; (ii) males aggregate at specific sites for display; (iii) the only resource females find on the lek is the male, i.e. the male genes; (iv) females can select her mate(s), although the necessity of this latter condition for lekking species has been highly debated. We applied these criteria to the endangered little bustard Tetrax tetrax, a species that is claimed to show an exploded lek mating system, but for which this has never been fully investigated. We monitored a population of little bustards in western France during 2 years to investigate the two central criteria in the assessment of their mating system: male aggregation in arenas and lack of consistent resources in male territories. We analysed the spatial distribution of little bustard male territories, the individual variation in size, and the land use characteristics of male territories, with particular attention to the habitats that may be considered as defensible resources. Displaying males showed an aggregated spatial distribution over the study area during the 2 years of survey. Male territories were rather large (19+/-16 ha), but a large among-male variability in territory size was observed. Land use within the territories included mainly permanent and semi-permanent crops. The variability in land use among territories suggests also that resources found within male territories were selected according to male needs (food and display) rather than to female needs (permanent crops that are more appropriate for reproduction). The mating system of the little bustard seems to match the general (and extended) definition of leks, at least in some populations. However, limits between resource defence polygyny and extreme exploded or resource-based leks are thin and unclear, and the little bustard is a good example of how lek definitions may be difficult to apply in non clear-cut empirical situations.     

Composition of free and adherent ruminal bacteria: inaccuracy of the microbial nutrient supply estimates obtained using free bacteria as reference samples and (15)N as the marker

Previous studies have indicated that (15)N enrichment of solid-associated bacteria (SAB) may be predicted from the same value in liquid-associated bacteria (LAB). The aims of this study were to confirm this and to measure the error in the nutrient supply from SAB, when LAB are used as the reference sample. For this purpose, the chemical and amino acid (AA) compositions of both the bacterial populations were studied in four experiments carried out on different groups of three rumen cannulated wethers. Diets (one in Experiments 1 and 4 and three in Experiments 2 and 3) had forage-to-concentrate ratios (dry matter (DM) basis) between 2 : 1 and 40 : 60, and were consumed at intake levels between 40 and 75 g DM/kg (BW)(0.75). The bacteria samples were isolated after continuous infusion of ((15)NH(4))(2)SO(4) (40, 18, 30 and 25 mg (15)N/day, in Experiments 1 to 4, respectively) for at least 14 days. In all experiments, SAB had consistently higher concentrations of organic matter (826 v. 716 g/kg DM, as average) and total lipids (192 v. 95 g/kg DM, as average) than LAB. Similar CP concentrations of both populations were observed, except a higher concentration in SAB than in LAB in Experiment 3. A consistent (in Experiment 4 only as tendency) higher AA-N/total N ratio (on average 17.5%) was observed in SAB than in LAB. The (15)N enrichment in SAB was systematically lower than in LAB. On the basis of the results of all studies a close relationship was found between the (15)N enrichment in SAB and LAB, which was shown irrespective of experiments. This relationship was established from Experiments 1 and 2 and the above cited previous results (n = 20; P < 0.001; R(2) = 0.996), and then confirmed from the results of Experiments 3 and 4. These relationships between SAB and LAB demonstrate that CP supply from SAB is underevaluated by, on average, 21.2% when LAB are used as the reference. This underevaluation was higher for true protein and even higher for the lipid supply (32.5% and 59.6%, respectively, as an average of the four experiments). Large differences in AA profile were observed between SAB and LAB. The prediction equation obtained using (15)N as the marker may be used to correct the errors associated with the traditional use of LAB as the reference sample, and therefore to obtain more accurate estimates of the microbial nutrient supply to the ruminants.     

Contrasting heterozygosity-fitness correlations between populations of a self-compatible shrub in a fragmented landscape

The mechanisms underlying heterozygosity-fitness correlations (HFCs) are subject of intense debates, especially about how important population features such as size or degree of isolation influence HFCs. Here, we report variation in HFCs between Large and Small populations of a self-compatible shrub (Myrtus communis) occurring within an extremely fragmented landscape. In each of the five study populations, we obtained data on both heterozygosity and fitness for 9-12 maternal families (i.e. offspring from the same mother plant). Whereas heterozygosity explained most of the variance (60-86?%) in growth rate of seedling families within Large populations, this relationship was absent within Small populations. Our results suggest that inbreeding may explain the observed HFCs within Large populations, and that different genetic processes (such as genetic drift and/or selection) could have overridden HFCs within Small populations. While it is difficult to draw general conclusions from five populations, we think our results open new research perspectives on how different genetic processes underlie variation in HFCs under different population contexts. Our study also points to a need for further attention on the complex relationships between heterozygosity in self-compatible plants and their progeny in relation to mating system variation. Finally, our results provide interesting new insights into how population genetic diversity is maintained or lost in a highly fragmented landscape.     

Presence of a large β(1-3)glucan linked to chitin at the Saccharomyces cerevisiae mother-bud neck suggests involvement in localized growth control

Previous results suggested that the chitin ring present at the yeast mother-bud neck, which is linked specifically to the nonreducing ends of β(1-3)glucan, may help to suppress cell wall growth at the neck by competing with β(1-6)glucan and thereby with mannoproteins for their attachment to the same sites. Here we explored whether the linkage of chitin to β(1-3)glucan may also prevent the remodeling of this polysaccharide that would be necessary for cell wall growth. By a novel mild procedure, β(1-3)glucan was isolated from cell walls, solubilized by carboxymethylation, and fractionated by size exclusion chromatography, giving rise to a very high-molecular-weight peak and to highly polydisperse material. The latter material, soluble in alkali, may correspond to glucan being remodeled, whereas the large-size fraction would be the final cross-linked structural product. In fact, the β(1-3)glucan of buds, where growth occurs, is solubilized by alkali. A gas1 mutant with an expected defect in glucan elongation showed a large increase in the polydisperse fraction. By a procedure involving sodium hydroxide treatment, carboxymethylation, fractionation by affinity chromatography on wheat germ agglutinin-agarose, and fractionation by size chromatography on Sephacryl columns, it was shown that the β(1-3)glucan attached to chitin consists mostly of high-molecular-weight material. Therefore, it appears that linkage to chitin results in a polysaccharide that cannot be further remodeled and does not contribute to growth at the neck. In the course of these experiments, the new finding was made that part of the chitin forms a noncovalent complex with β(1-3)glucan.     

Yellow fever vector live-virus vaccines: West Nile virus vaccine development

By combining molecular-biological techniques with our increased understanding of the effect of gene sequence modification on viral function, yellow fever 17D, a positive-strand RNA virus vaccine, has been manipulated to induce a protective immune response against viruses of the same family (e.g. Japanese encephalitis and dengue viruses). Triggered by the emergence of West Nile virus infections in the New World afflicting humans, horses and birds, the success of this recombinant technology has prompted the rapid development of a live-virus attenuated candidate vaccine against West Nile virus.     

Rendimiento diagnóstico de los hallazgos ecográficos de los nódulos tiroideos en la detección de lesiones malignas

ObjetiveTo evaluate the diagnostic efficiency of sonographic findings (nodule size, the presence of microcalcifications and echogenicity) compared with the results of fine-needle aspiration biopsy (FNAB) of thyroid nodules.MethodsThe results of cytology and the ultrasound characteristics of 341 thyroid nodules were analyzed.ResultsA total of 25.5% of the FNAB were inadequate, 65.1% were benign and 7.9% were suspicious or malignant. The percentage of inadequate samples was clearly larger in nodules smaller that 15 mm but that of malignant cytologies was also larger in this group of nodules (11.1 versus 2.8%: p=0.04). The percentage of inadequate samples among nodules smaller than 10 mm was 66.7% and no malignancies were detected. A notable finding was the absence of suspicious or malignant FNAB in hyperechogenic and anechogenic nodules. In contrast, the echogenic feature most frequently associated with malignancy was hypoechogenicity, although echogenicity, as a whole, was not significantly associated with malignancy (p=0.313). Most cases with microcalcifications (6 of 10) were malignant compared with four of the 239 nodules (1.7%) without calcifications (p<0.001). Multivariate logistic regression revealed that the only variable maintaining a significant association with malignancy was the presence of microcalcifications.ConclusionsThyroid cytology is an efficient method to evaluate thyroid nodules larger than 10 mm. The presence of nodule microcalcifications is significantly associated with malignancy, while hyperechogenicity and anechogenicity are associated with benign nodules.