Hyoid bone morphology in patients with isolated robin sequence - A case-control study utilizing 3D morphable models.

Background: Abnormalities of the hyoid bone are associated with impairment of oropharyngeal functions including feeding, swallowing, and breathing. Few studies have characterized anatomic abnormalities of the hyoid in patients with Robin sequence (RS), e.g. a less mineralized and voluminous hyoid. The purpose of this study was to compare normal hyoid bone morphology and hyoid bone morphology in children with isolated RS. Methods: Three-dimensional (3D) reconstructions of the hyoid bone were obtained from CT-imaging of children with RS and unaffected controls. A 3D morphable model was constructed using Principal Component Analysis (PCA). Partial least squares - Discriminant Analysis (PLS-DA) and multivariate analysis of variance (MANOVA) were used to characterize and compare hyoid shape differences between patients with RS and an age-matched control group. Results: The study included 23 subjects with RS (mean age 9.8 ± 10.3 months) and 46 age-matched control samples. A less voluminous hyoid was observed for the RS group with a larger lateral divergence of the greater horns compared to controls (MANOVA, p-value<0.001). The first shape variable from the PLS-DA model showed a significant correlation for the observed variance between the two groups (Spearman R = -0.56, p-value<0.001). The control samples and 151 CT-scans of subjects up to age 4 years were used to create a 3D morphable model of normal hyoid shape variation (n = 197, mean age 22.1 ± 13.1 months). For the normal 3D morphable model, a high degree of allometric shape variation was observed along the first principal component. Conclusions: The 3D morphable models provide a comprehensive and quantitative description of variation in normal hyoid bone morphology, and allow detection of distinct differences between patients with isolated RS and controls.

Developmental regulation of DNA replication timing at the human beta globin locus.

The human beta globin locus replicates late in most cell types, but becomes early replicating in erythroid cells. Using FISH to map DNA replication timing around the endogenous beta globin locus and by applying a genetic approach in transgenic mice, we have demonstrated that both the late and early replication states are controlled by regulatory elements within the locus control region. These results also show that the pattern of replication timing is set up by mechanisms that work independently of gene transcription.

Protection against pulmonary infection with Pseudomonas aeruginosa following immunization with P. aeruginosa-pulsed dendritic cells.

To develop a Pseudomonas aeruginosa vaccine that allows the host immune system to select the antigens, we hypothesized that dendritic cells (DC) pulsed with P. aeruginosa would induce protective immunity against pulmonary infections with P. aeruginosa. Incubation of murine bone marrow-derived DC with P. aeruginosa in vitro led to uptake of P. aeruginosa and activation of the DC. Spleen-derived CD4(+) cells from mice immunized with P. aeruginosa-pulsed DC showed increased proliferation, demonstrating that DC pulsed with P. aeruginosa were capable of eliciting a P. aeruginosa-specific immune response. To evaluate if P. aeruginosa-pulsed DC can induce protective immunity against P. aeruginosa pulmonary infection, DC incubated with P. aeruginosa in vitro were administered systemically to syngeneic mice, and the mice were then challenged by intrapulmonary infection with P. aeruginosa (5 x 10(4) CFU/mouse) 13 days later. Unimmunized control mice and mice who had previously received naive DC or DC stimulated with lipopolysaccharide or Escherichia coli died within 72 h. In contrast, 45% of mice receiving P. aeruginosa-pulsed DC demonstrated prolonged survival (>14 days). Finally, DC-pulsed with heat-inactivated P. aeruginosa protected CD8(-/-) but not CD4(-/-) mice, demonstrating that CD4(+) T cells were required for the DC pulsed with P. aeruginosa to induce protective immunity.

Noninvasive evaluation of chronic venous insufficiency. Use of foot mercury strain-gauge plethysmography.

Twenty-seven patients (40 extremities) with chronic venous insufficiency and 30 controls (60 extremities) were studied using noninvasive measurements of foot volume (foot mercury strain-gauge plethysmography [FMSGP]). Tests were performed with exercise (sitting) and with elevation (Trendelenburg), with and without tourniquets, to evaluate muscle pump efficiency and valvular competence. Empiric venous sufficiency indexes were calculated for exercise and for Trendelenburg's test. Test results were compared with invasive ambulatory venous pressure (AVP) data (13) and with ascending and retrograde venography (25). Distinctive plethysmographic patterns revealed the insufficiency was saphenous (four), calf perforators (14), saphenous plus perforators (seven), and deep venous (15), including occlusion (four). Correlation with AVP and venography was good but FMSGP was more discriminating, providing precise anatomical information, better reproducibility, and distinguishing poor muscle pump function from regurgitation. Patients accept FMSGP well. Repeated postoperative data are readily obtained for evaluation of surgical procedures.

Factors modulating the activity of ornithine decarboxylase in rat HTC cells.

Dibutyryl cyclic AMP (dibu-cAMP), N1-acetylputrescine and both N1-acetylspermidine and N8-acetylspermidine regularly induce ornithine decarboxylase activity in both a time- and a concentration-dependent manner when added to HTC cells in suspension culture. N,N'-Diacetyl-1,6-diaminohexane and N-acetyl-1,3-diaminopropane, aliphatic analogues of acetylated putrescine containing the carboxamide bond but not known to occur naturally in animal tissues, are also effective inducing agents. The addition of 10(-2)M putrescine to the cell culture at the time of maximal induction of ODC in HTC cells by dibu-cAMP causes a marked decrease in the apparent half-life of ODC in less than 10 minutes. Ten-fold concentration of HTC cells at the time of their maximal induction by dibu-cAMP also results in an immediate decrease both in the specific activity of ODC and in its apparent half-life. When the population density of induced HTC cells is increased the relative effect of exposure of cells to increasing levels of putrescine is lessened. Some of the implications of these observations are discussed.