Sexual conflict in Gerris gillettei (Insecta: Hemiptera): intraspecific intersexual correlated morphology and experimental assessment of behaviour and fitness.

The contemporary dynamics of sexually antagonistic coevolution caused by sexual conflicts have seldom been investigated at the intraspecific level. We characterized natural populations of Gerris gillettei and documented significant intersexual correlations for morphological traits previously related to sexual conflict in water striders. These results strongly indicate that sexually antagonistic coevolution contributed to population differentiation and resulted in different balances of armaments between the sexes within natural populations of this species. No-choice mating experiments further revealed that both male and male-female relative arms levels influence copulation duration. However, there were no asymmetries in reproductive behaviour and fitness between sympatric and allopatric mating pairs, suggesting that differentiation by sexual conflict was not sufficient to influence the outcome of mating interactions. Altogether, these results question the relative importance of female connexival spines vs. genitalia traits in mediating pre- and post-copulatory conflict in Gerris.

The determinant role of temporary proglacial drainages on the genetic structure of fishes.

Phylogeographic studies have shed light on Pleistocene glaciations as a key factor in shaping present-day genetic structure of many organisms. In formerly glaciated regions, the combined action of several factors such as refuges origin, physiological capacities and demographic parameters have contributed importantly to this process but specifically for each species. Therefore, a fine-scale genetic structure is not expected to be similar for different species, unless it has been modulated by the action of a strong environmental pressure. The aim of this study is to investigate the effects of postglacial environment on the genetic structure of fishes. To achieve this objective, three fish species (northern pike, lake whitefish and yellow perch) commonly found in sympatry in Laurentian Shield lakes but displaying different ecological and physiological characteristics were analysed. The comparison of these unrelated species was performed to identify the factors determining the organization of their genetic structure. Populations of all species mostly originated from the Mississippian refuge. Low genetic differentiation was observed among populations but significant structures were detected for the three species. Despite marked differences among species, these structures presented common characteristics: a lack of congruence with drainage and a longitudinal organization. This suggested that the dispersion of species occurred independently, leading to a species-specific structure. However, the settling of populations appeared to be mediated by a dynamic system of proglacial meltwater streams associated to the glacial Lake Ojibway-Barlow, providing such similarities among species.

Single-point mutations at the surface of MB-1Trp lead to important changes in its conformational properties.

Protein design is currently used for the creation of new proteins with desirable traits. In our lab we focus on the synthesis of proteins with high essential amino acid content having potential applications in animal nutrition. One of the limitations we face in this endeavour is achieving stable proteins despite a highly biased amino acid content. We report here the synthesis and the characterization of three variants of MB-1Trp in which two solvent-exposed Leu have been replaced by Glu allowing for the formation of new salt bridges at the surface of the protein. Although both mutations were expected to be similar (i.e. same mutation in a comparable local environment), they appear to have different effects on MB-1Trp as shown by far-UV circular dichroism, thermal denaturation, fluorescence and proteolytic resistance measurements. For the mutation Leu68Glu, an increase in the protein melting temperature of 6 degrees C was observed. Surprisingly, the mutation in position Leu19Glu led to a decrease in melting temperature and a modification of tertiary structure.

Design of high essential amino acid proteins: two design strategies for improving protease resistance of the nutritious MB-1 protein.

Protein design is currently used for the creation of new proteins with desirable traits. In our lab, we focus on the synthesis of proteins with high essential amino acid content having potential applications in animal nutrition. One of the limitations we face in this endeavour is achieving stable proteins despite a highly biased amino acid content. We report here the synthesis and characterisation of two mutants derived from our MB-1 designer protein. The first mutant contains a disulphide bridge designed to cross-link remote segments of the polypeptide chain. The second one is a Tyr62-Trp mutant, where position 62 is buried in the core of the protein. Both mutants were found to be largely helical as per design, and based on thermal denaturation experiments, were substantially more stable than the MB-1 parent molecule. Enhancement of conformational stability in MB-1Trp translated into an impressive improvement of its ability to resist proteolytic degradation. Furthermore, digestion experiments intended to model degradation of proteins in a cow's rumen revealed that MB-1Trp's resistance to degradation compared to that of cytochrome c. Design strategies used for these mutants are discussed with regards to their applicability in creating efficient nutritional proteins.

Replacement of tyr62 by trp in the designer protein milk bundle-1 results in significant improvement of conformational stability.

Protein design is currently used for the creation of new proteins with desirable traits. In our lab, we focus on the synthesis of proteins with high essential amino acid content, having potential application in animal nutrition. One of the limitations we face in this endeavor is the achievement of stable proteins in spite of a highly biased amino acid content. We report here the synthesis and characterization of MB-1Trp, a protein with a tailored content in selected essential amino acids. The protein is a Tyr62-Trp mutant of the parent molecule MB-1 described earlier. The new protein is largely helical as per design, is well folded, and has a melting temperature of 55 degrees C. Its resistance to proteolytic degradation compares to that of cytochrome c, a protein of similar size. Design strategy used for MB-1Trp is discussed with regards to its applicability toward the creation of efficient nutritional proteins.

Do proficiency test results correlate with the work performance of screeners who screen Papanicolaou smears?

We rescreened Papanicolaou smear slides from 40,245 women, which had been examined by 81 cytology screeners, scored the screeners' work performance, and compared these scores with the results of the screeners' performance on glass slide and computer-based proficiency tests. All diagnoses (i.e., from the proficiency tests, the original slides, and the rescreened slides) were classified in the 4 diagnostic categories specified in the Clinical Laboratory Improvement Amendments. The rescreening scores were standardized to account for different distributions of abnormalities in the proficiency tests and rescreened slides. We compared a standardized score with the proficiency test scores. Of the cases, 91% were categorized as normal, benign, or reactive changes when rescreened, and 98% of these agreed with the original diagnosis. Sixteen percent of low-grade and 15% of high-grade intraepithelial lesions were classified as normal. The rank correlation between the rescreening scores and both proficiency tests was 0.24 using a scoring scheme for cytotechnologists. The correlation between the rescreening and proficiency testing scores indicates that performance on a 10-slide test gives some indication of the true performance of screeners. The computer-based test shows promise as an alternative to the glass slide test but needs further development and validation.

Ontogeny and species differences in the pancreatic expression and localization of the CCK(A) receptors.

We have evaluated the presence and localization of the CCK(A) receptor in rat, mouse, pig and human fetal pancreas by Northern, Western blots and immunofluorescence techniques. In the rat, parallelism exists between development of the CCK(A) receptor mRNA and protein with maximal peaks of expression during the suckling period. In the course of pancreatitis induction, CCK(A) receptor mRNA were maximally expressed and sustained during the gland's regeneration. In the rat and mouse pancreas, the CCK(A) receptor protein is localized around the acinar cells and beta cells of the islets of Langerhans. In the adult pig and fetal human pancreas, the CCK(A) receptor proteins were detected by Western blot. By immunofluorescence, its detection was possible only in the islet of Langerhans of the pig pancreas. These new findings support the views that CCK plays important and various roles in specific physiological systems of the pancreas of different species.

Linoleic acid transport by human placental syncytiotrophoblast membranes.

The placenta syncytiotrophoblast is the site of exchange of nutrients, lipids and minerals between the mother and the fetus. In order to characterize the transport of fatty acids by the placenta, we purified bipolar syncytiotrophoblast brush border and basal plasma membranes from human placenta. These purified brush border and basal plasma membranes enriched 3-fold and 22-fold, respectively, in sodium/potassium-ATPase and 27-fold and 6-fold in alkaline phosphatase activity, compared with the placental homogenates. Fatty acid transport was performed at different fatty acid/albumin ratios to evaluate the optimal uptake conditions. The maximal transport efficiency, for linoleic acid bound to albumin by sonication, was obtained with a 6:1 fatty acid/albumin ratio in brush border and basal plasma membranes. The linoleic acid transport observed with brush border membranes followed Michaelis-Menten kinetics, with a Michaelis constant of 7.89 +/- 0.01 microM and a maximal incorporation rate of 30.80 +/- 6.39 pmol.mg-1.min-1. Linoleic acid transport was very low in basal plasma membranes and we obtained a Michaelis constant of 0.95 +/- 0.01 microM and a maximal incorporation rate of 1.62 +/- 5.06 pmol.mg-1.min-1. In order to show that linoleic acid accumulated within brush border and plasma membrane vesicles, and to eliminate the possibility of a non-specific binding of fatty acid to these membranes, we demonstrated by an osmolarity experiment, the decrease of the linoleic acid transport in brush border and basal plasma membranes obtained in the presence of 455 microM essential fatty acid at 23 degrees C for 180 min. The results presented in this study suggest that linoleic acid is transported significantly by syncytiotrophoblast brush border membranes and basal plasma membranes. Thus, it may represent a unidirectional transport from mother to fetus through the brush border membranes facing the mother, followed by transport at a slower rate through basal plasma membranes facing the fetus.