Sexual dimorphism in human facial expressions by 3D surface processing.

Human face is a dynamic system where facial expressions can rapidly modify geometry of facial features. Facial expressions are believed to be universal across world populations, but only a few studies have explored whether grimacing is sexually dimorphic and if so to what extent. The present paper explores inter- and intra-individual variation of human facial expressions with respect to individual's sex based on a set of neutral and expression-varying 3D facial scans. The study sample composed of 20 individuals (10 males and 10 females) for whom 120 scans featuring grimaces associated with disgust, surprise, "u" sound, smile and wide smile were collected by an optical scanner Vectra XT. In order to quantify the dissimilarity among 3D images, surface comparison approach based on aligned 3D meshes and closest point-to-point distances was carried out in Fidentis Analyst application. The study revealed that sexual dimorphism was indeed one of the factors which determined the extent and characteristics of facial deformations recorded for the studied expressions. In order to produce a grimace, males showed a tendency towards extending their facial movements while females were generally more restrained. Furthermore, the facial movements linked to the wide smile and "u" sound were revealed as the most extensive relative to the other expressions, while the smile and surprise were shown indistinguishable from the neutral face.

The morphology of human hyoid bone in relation to sex, age and body proportions.

Morphological aspects of the human hyoid bone are, like many other skeletal elements in human body, greatly affected by individual's sex, age and body proportions. Still, the known sex-dependent bimodality of a number of body size characteristics overshadows the true within-group patterns. Given the ambiguity of the causal effects of age, sex and body size upon hyoid morphology the present study puts the relationship between shape of human hyoid bone and body proportions (height and weight) under scrutiny of a morphological study. Using 211 hyoid bones and landmark-based methods of geometric morphometrics, it was shown that the size of hyoid bones correlated positively with measured body dimensions but showed no correlation if the individual's sex was controlled for. For shape variables, our results revealed that hyoid morphology is clearly related to body size as expressed in terms of the height and weight. Yet, the hyoid shape was shown to result primarily from the sex-related bimodal distribution of studied body size descriptors which, in the case of the height-dependent model, exhibited opposite trends for males and females. Apart from the global hyoid shape given by spatial arrangements of the greater horns, body size dependency was translated into size and position of the hyoid body. None of the body size characters had any impact on hyoid asymmetry. Ultimately, sexually dimorphic variation was revealed for age-dependent changes in both size and shape of hyoid bones as male hyoids tend to be more susceptible to modifications with age than female bones.

Effects of fluorescent pseudo-ATP and ATP-metal analogs on secondary structure of Na(+)/K(+)-ATPase.

The secondary structure of Na(+)/K(+)-ATPase after modification of the ATP-binding sites was analyzed. Consistently with recent reports, we found in trypsin-treated Na(+)/K(+)-ATPase additionally to alpha-helix also beta-sheet structures in the transmembrane segments. However, binding of fluorescein 5'-isothiocyanate (FITC), the pseudo-ATP analog, to the ATP-binding site did not affect the secondary structure of undigested Na(+)/K(+)-ATPase. Consequently, fluorescence intensity changes of FITC-labeled Na(+)/K(+)-ATPase commonly used to observe conformational transitions of the enzyme reflect physiological changes of the native structure. The metal complex analogues of ATP, Cr(H(2)O)(4)ATP and Co(NH(3))(4)ATP, on the other hand, affected the secondary structure of Na(+)/K(+)-ATPase. We propose that these changes in the secondary structure are responsible for inhibition of backdoor phosphorylation.

Molecular distance measurements reveal an (alpha beta)2 dimeric structure of Na+/K+-ATPase. High affinity ATP binding site and K+-activated phosphatase reside on different alpha-subunits.

ATP hydrolysis by Na+/K+-ATPase proceeds via the interaction of simultaneously existing and cooperating high (E1ATP) and low (E2ATP) substrate binding sites. It is unclear whether both ATP sites reside on the same or on different catalytic alpha-subunits. To answer this question, we looked for a fluorescent label for the E2ATP site that would be suitable for distance measurements by Förster energy transfer after affinity labeling of the E1ATP site by fluorescein 5'-isothiocyanate (FITC). Erythrosin 5'-isothiocyanate (ErITC) inactivated, in an E1ATP site-blocked enzyme (by FITC), the residual activity of the E2ATP site, namely K+-activated p-nitrophenylphosphatase in a concentration-dependent way that was ATP-protectable. The molar ratios of FITC/alpha-subunit of 0.6 and of ErITC/alpha-subunit of 0.48 indicate 2 ATP sites per (alpha beta)2 diprotomer. Measurements of Förster energy transfer between the FITC-labeled E1ATP and the ErITC-labeled or Co(NH3)4ATP-inactivated E2ATP sites gave a distance of 6.45 +/- 0.64 nm. This distance excludes 2 ATP sites per alpha-subunit since the diameter of alpha is 4-5 nm. Förster energy transfer between cardiac glycoside binding sites labeled with anthroylouabain and fluoresceinylethylenediamino ouabain gave a distance of 4.9 +/- 0.5 nm. Hence all data are consistent with the hypothesis that Na+/K+-ATPase in cellular membranes is an (alpha beta)2 diprotomer and works as a functional dimer (Thoenges, D., and Schoner, W. (1997) J. Biol. Chem. 272, 16315-16321).

Oligosaccharide organization of the beta-subunits of pig kidney Na+/K+-ATPase.

The distance between the beta-subunits of Na+/K+-ATPase isolated from pig dark red kidney medulla was determined by Förster energy transfer. First, oligosaccharides of the beta-subunit were shown to be labelled with three fluorophores: Lucifer yellow (LY), Lissamine rhodamine B sulfonyl hydrazine (LRSH) and Cascade blue (CB). Further, LY and LRSH were used as the donor and the acceptor, respectively, for Förster energy transfer studies to determine the localization of the beta-subunit in the native enzyme which is known to be formed as a tetramer (alphabeta)2. It was found that the beta-subunits in the functional enzyme complex in the membrane are not localized next to each other but are spatially separated. The distance between fluorophores covalently attached to the beta-subunits was found to be 5.1 nm. This conclusion was confirmed by measurements with another donor-acceptor pair CB-LY. The results also support the idea of a direct interaction of the beta-subunit with the extracellular part of the alpha-subunit. These interactions were modified in the presence of millimolar concentrations of magnesium ions. This indicates a crucial role of magnesium in extracellular interactions between the alpha and beta subunits.

Quenching of 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole-modified Na+/K+-ATPase reveals a higher accessibility of the low-affinity ATP-binding site.

7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) labeled Na+/K+-ATPase covalently with two different inactivation constants (Ki = 2.5 microM; Ki' = 10 microM). It apparently modified the two different ATP-binding sites of the enzyme since it decreased the activity of the E2ATP site, i.e. the K+-activated para-nitrophenylphosphatase activity, in an enzyme whose high-affinity E1ATP site had been blocked by fluorescein 5'isothiocyanate (FITC). It also reduced the activity of the E1ATP site, i.e. the Na+-activated protein phosphorylation, in an enzyme whose low-affinity E2ATP site had been blocked by Co(NH3)4PO4. Fluorescence quenching experiments with KI, CsCl and MnCl2 of the NBD-Cl-labeled Na+/K+-ATPase revealed two differently accessible types of fluorophores depending on the ATP site: The E2ATP site apparently differs from the E1ATP site in that it is more open because the fluorophore labeling in the E2ATP site was sterically better accessible for quenchers.

Possible nystatin-protein interaction in yeast plasma membrane vesicles in the presence of ergosterol. A Förster energy transfer study.

The Förster energy transfer from tryptophan residues of membrane proteins to nystatin was measured in reconstituted yeast plasma membrane vesicles free of, or doped with, ergosterol. We wanted to elucidate whether the functional change of membrane transport proteins from H+ symporters to facilitators, observed in ergosterol-containing plasma membrane vesicles on addition of nystatin [Opekarová and Tanner (1994) FEBS Lett. 350, 46-50], is reflected in altered protein-nystatin relations within the membrane. Both frequency-domain and time-domain time-resolved fluorescence spectroscopy showed that in the presence of ergosterol nystatin is located much closer to membrane proteins than in its absence.