Calcaneus ultrasound measurements in a convenience sample of healthy youth.

We examined age-related changes in quantitative ultrasound of the calcaneus in 311 healthy males and females ages 6.6-20 yr using the Lunar Achilles ultrasound device. This equipment has been adapted for pediatric use with the provision of shims designed to properly position smaller feet relative to the transducers. Broadband ultrasound attenuation (BUA) (decibels/megahertz), speed of sound (SOS) (meters/second), and stiffness index (SI) (percent) increased across the age range until a plateau was reached at 16-18 yr. BUA increased by 40%, SOS by 4%, and SI by 80% across this age range. There was no gender difference in age-related gains. Age, weight, height, and hours of weight-bearing physical activity were all significantly associated with BUA, SOS, and SI. After controlling for age and weight, hours of weight-bearing physical activity showed little to no additional effect on these parameters. Short-term in vivo precision using this device was similar in children to that observed in adults in our laboratory; coefficients of variation for between-day measurements were 1.8, 0.6, and 3.2% for BUA, SOS, and SI, respectively. These data support the feasibility of using the Lunar Achilles in evaluating pediatric bone mass. The ability of this technique to discriminate between osteopenic and normal children remains to be determined.

Targeting of zyxin to sites of actin membrane interaction and to the nucleus.

The localization of proteins to particular intracellular compartments often regulates their functions. Zyxin is a LIM protein found prominently at sites of cell adhesion, faintly in leading lamellipodia, and transiently in cell nuclei. Here we have performed a domain analysis to identify regions in zyxin that are responsible for targeting it to different subcellular locations. The N-terminal proline-rich region of zyxin, which harbors binding sites for alpha-actinin and members of the Ena/VASP family, concentrates in lamellipodial extensions and weakly in focal adhesions. The LIM region of zyxin displays robust targeting to focal adhesions. When overexpressed in cells, the LIM region of zyxin causes displacement of endogenous zyxin from focal adhesions. Upon mislocalization of full-length zyxin, at least one member of the Ena/VASP family is also displaced, and the organization of the actin cytoskeleton is perturbed. Zyxin also has the capacity to shuttle between the nucleus and focal adhesion sites. When nuclear export is inhibited, zyxin accumulates in cell nuclei. The nuclear accumulation of zyxin occurs asynchronously with approximately half of the cells exhibiting nuclear localization of zyxin within 2.3 h of initiating leptomycin B treatment. Our results provide insight into the functions of different zyxin domains.

Adenomatous polyposis coli protein contains two nuclear export signals and shuttles between the nucleus and cytoplasm.

Mutational inactivation of the adenomatous polyposis coli (APC) tumor suppressor initiates most hereditary and sporadic colon carcinomas. Although APC protein is located in both the cytoplasm and the nucleus, the protein domains required to maintain a predominantly cytoplasmic localization are unknown. Here, we demonstrate that nuclear export of APC is mediated by two intrinsic, leucine-rich, nuclear export signals (NESs) located near the amino terminus. Each NES was able to induce the nuclear export of a fused carrier protein. Both APC NESs were independently able to interact with the Crm1 nuclear export factor and substitute for the HIV-1 Rev NES to mediate nuclear mRNA export. Both APC NESs functioned within the context of APC sequence: an amino-terminal APC peptide containing both NESs interacted with Crm1 and showed nuclear export in a heterokaryon nucleocytoplasmic shuttling assay. Also, mutation of both APC NESs resulted in the nuclear accumulation of the full-length, approximately 320-kDa APC protein, further establishing that the two intrinsic APC NESs are necessary for APC protein nuclear export. Moreover, endogenous APC accumulated in the nucleus of cells treated with the Crm1-specific nuclear export inhibitor leptomycin B. Together, these data indicate that APC is a nucleocytoplasmic shuttle protein whose predominantly cytoplasmic localization requires NES function and suggests that APC may be important for signaling between the nuclear and cytoplasmic compartments of epithelial cells.

Nuclear-cytoplasmic shuttling of the focal contact protein, zyxin: a potential mechanism for communication between sites of cell adhesion and the nucleus.

Integrin-dependent cell adhesion to specific extracellular matrix molecules has been demonstrated to trigger dramatic changes in gene expression that can affect cell fate. However, little is understood about the molecular mechanism by which events at sites of cell- substratum adhesion are communicated to the cell interior to regulate the transcriptional apparatus. By analogy to classical mechanisms of cell surface receptor function, it seems likely that some components of the integrin-activated signal transduction machinery will be colocalized with cell adhesion molecules. Zyxin is a low abundance phosphoprotein that accumulates with integrins at sites of cell-substratum attachment. Here we show that zyxin exhibits a functional nuclear export signal that is required to keep zyxin concentrated in the cytoplasm and is sufficient to direct nuclear proteins to the cytosol. Furthermore, we demonstrate that native zyxin shuttles between the nucleus and sites of cell adhesion in fibroblasts and is thus an excellent candidate for relaying information between these two compartments.

Two cross-linguistic factors underlying tongue shapes for vowels.

Desirable characteristics of a vocal-tract parametrization include accuracy, low dimensionality, and generalizability across speakers and languages. A low-dimensional, speaker-independent linear parametrization of vowel tongue shapes can be obtained using the PARAFAC three-mode factor analysis procedure [Harshman et al., J. Acoust. Soc. Am. 62, 693-707 (1977)]. Harshman et al. applied PARAFAC to midsagittal x-ray vowel data from five English speakers, reporting that two speaker-independent factors are required to accurately represent the tongue shape measured along anatomically normalized vocal-tract diameter grid lines. Subsequently, the cross-linguistic generality of this parametrization was brought into question by the application of PARAFAC to Icelandic vowel data, where three nonorthogonal factors were reported [Jackson, J. Acoust. Soc. Am. 84, 124-143 (1988)]. This solution is shown to be degenerate; a reanalysis of Jackson's Icelandic data produces two factors that match Harshman et al.'s factors for English vowels, contradicting Jackson's distinction between English and Icelandic language-specific "articulatory primes". To obtain vowel factors not constrained by artificial measurement grid lines, x-ray tongue shape traces of six English speakers were marked with 13 equally spaced points. PARAFAC analysis of this unconstrained (x,y) coordinate data results in two factors that are clearly interpretable in terms of the traditional vowel quality dimensions front/back, high/low.

Nifedipine blocks retention of a visual discrimination task in chicks.

Reports that the dihydropyridine Ca2+ channel antagonists may facilitate memory led to the present study of the behavioral effects of nifedipine. Ninety-two 4-day-old male chicks received 0, 100 nM, 100 microM, or 10 mM nifedipine. Drugs were administered in volumes of 2 microliters into the fourth cerebral ventricle 5 min before training on a visual discrimination task. Nifedipine did not produce any detectable changes in behavior during acquisition trials. Retention, however, was impaired 24 h after training in the 100 nM and 100 microM nifedipine-treated chicks, which made significantly more errors than controls. Nifedipine did not affect the amount of time required to complete the task. No effects on body weight gain were detected, suggesting that the memory impairment was not due to a change in feeding behavior. These data are discussed in terms of the role of calcium-dependent processes in memory.