AhR, ARNT, and CYP1A1 mRNA quantitation in cultured human embryonic palates exposed to TCDD and comparison with mouse palate in vivo and in culture.
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is developmentally toxic in many species and induces cleft palate in the C57BL/6N mouse embryo. Palatogenesis in mouse and human embryos involves homologous processes at the morphological, cellular, and molecular levels. In organ culture, mouse and human palates respond similarly to TCDD. The present study quantitates the expression of AhR, ARNT, and CYP1A1 mRNA in human embryonic palates in organ culture. Palatal tissues were exposed to 1 x 10(-10), 1 x 10(-9), or 1 x 10(-8) M TCDD or control medium and sampled at 0, 2, 4, and 6 hours for quantitative RT-PCR using a synthetic RNA internal standard. Similar measurements of CYP1A1 gene expression were collected for mouse palates cultured in this model. In human palates, AhR expression correlated with ARNT and CYP1A1 mRNA expression. TCDD induction of CYP1A1 was time- and concentration-dependent. The expression of these genes presented a uniform and continuous distribution across the group of embryos, with no subset of either high or low expressors/responders. The ratio of AhR to ARNT was approximately 4:1. AhR mRNA increased during the culture period in both treated and control subjects; however, ARNT expression was relatively constant. TCDD did not alter either AhR or ARNT expression in a consistent dose- or time-related manner. Comparison of human and mouse data showed a high correlation across species for the induction of CYP1A1. Human embryos expressed approximately 350 times less AhR mRNA than the mouse, and in earlier studies it was shown that human palates required 200 times more TCDD to produce the same effects. When the morphological, cellular, and molecular responses to TCDD between mouse and human are compared, it seems highly unlikely that human embryos could be exposed to sufficient TCDD to achieve changes in palatal differentiation that would lead to cleft palate. (+info)
RT-PCR quantification of AHR, ARNT, GR, and CYP1A1 mRNA in craniofacial tissues of embryonic mice exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin and hydrocortisone.
C57BL/6N mouse embryos exposed to hydrocortisone (HC) or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) develop cleft palate. An interaction between these agents produces clefts at doses which alone are not teratogenic. The glucocorticoid receptor (GR) and dioxin receptor (AhR) mediated these responses and their gene expression was altered by TCDD and/or HC in palates examined on gestation day (GD) 14 by Northern blot analysis and in situ hybridization. The present study quantifies AhR, AhR nuclear translocator (ARNT), and GR mRNA at 4, 12, 24, and 48 h after exposure (time 0 = dose administration at 8 A.M. on gestation day 12) on GD12 to TCDD (24 micrograms/kg), HC (100 mg/kg) or HC (25 mg/kg) + TCDD (3 micrograms/kg). The induction of CYP1A1 mRNA was also quantified at 2, 4, 6, 12, 24, and 48 h for control and TCDD-exposed samples. Total RNA was prepared from midfacial tissue of 4-6 embryos/litter at each time and dose. An RNA internal standard (IS) for each gene was synthesized, which included the gene's primer sequences separated by a pUC19 plasmid sequence. Reverse transcription-polymerase chain reaction (RT-PCR) was performed on total RNA + IS using a range of 5-7 IS concentrations across a constant level of total RNA. PCR products were separated in gels (mRNA and IS-amplified sequences differed by 30-50 bases), ethidium bromide-stained, imaged (Hamamatsu Photonics Systems, Bridgewater, NJ), and quantified with NIH Image. CYP1A1 mRNA was significantly induced in the TCDD-exposed samples at all time points examined (p = 0.005 at 2 h and 0.001 after 2 h). During palatal shelf outgrowth on GD12, AhR mRNA levels increased significantly and this was not affected by treatment with TCDD or HC + TCDD. A significant increase in GR was detected at 24 h (p < 0.05) and this was unaffected by any of the exposures. Expression of ARNT increased at 12 h (p < 0.001); however, treatment with HC or HC + TCDD blocked this increase (p < 0.05). At 24 h, the TCDD-treated embryos had significantly lower ARNT mRNA compared with controls (p < 0.001). The relative overall expression level of the genes was AhR > ARNT > GR. Within individuals, expression of AhR and/or ARNT was highly correlated with GR level. In conclusion, CYP1A1 mRNA was expressed in developing craniofacial tissue and was highly induced by TCDD exposure. AhR, ARNT, and GR mRNA are upregulated in early palatogenesis, although not on the same schedule. The TCDD-induced decrease in ARNT at 24 h after dosing and the HC and HC + TCDD-induced delay in upregulation of ARNT may affect the dynamics of heterodimer formation between AhR and ARNT. The changes in ARNT mRNA level could also affect availability of this transcriptional regulator to interact with other potential partners, and these effects, separately or in combination, may be involved in disruption of normal embryonic development. (+info)
Neck soft tissue and fat distribution: comparison between normal men and women by magnetic resonance imaging.
BACKGROUND: Obesity and increased neck circumference are risk factors for the obstructive sleep apnoea/hypopnoea syndrome (SAHS). SAHS is more common in men than in women, despite the fact that women have higher rates of obesity and greater overall body fat. One factor in this apparently paradoxical sex distribution may be the differing patterns of fat deposition adjacent to the upper airway in men and women. A study was therefore undertaken to compare neck fat deposition in normal men and women. METHODS: Using T1 weighted magnetic resonance imaging, the fat and tissue volumes in the necks of 10 non-obese men and 10 women matched for age (men mean (SE) 36 (3) years, women 37 (3) years, p = 0.7), body mass index (both 25 (0. 6) kg/m2, p>0.9), and Epworth Sleepiness Score (both 5 (1), p = 0.9) were assessed; all denied symptoms of SAHS. RESULTS: Total neck soft tissue volume was greater in men (1295 (62) vs 928 (45) cm3, p<0. 001), but the volume of fat did not differ between the sexes (291 (29) vs 273 (18) cm3, p = 0.6). The only regions impinging on the pharynx which showed a larger absolute volume of fat in men (3.2 (0. 7) vs 1.1 (0.3) cm3, p = 0.01) and also a greater proportion of neck fat in men (1.3 (0.3)% vs 0.4 (0.1)%, p = 0.03) were the anterior segments inside the mandible at the palatal level. CONCLUSIONS: There are differences in neck fat deposition between the sexes which, together with the greater overall soft tissue loading on the airway in men, may be factors in the sex distribution of SAHS. (+info)
The maintenance of rat palatal mucosa in organ culture.
Palatal mucosa from neonatal rats was maintained under organ culture conditions in a chemically defined medium for periods up to 28 days. The histological state of the cultured palatal mucosa was compared with that of control tissue from growing animals of comparable age. The control tissues showed an increase in epithelial thickness, first noticeable at 17 days. Whilst the general structure of the tissues in organ culture was preserved for the duration of the experiment, some changes in epithelial behavior were evident. There was an increase in epithelial thickness up to 6 days, followed by a reduction in the nucleated cell layer of the epithelium to a thickness comparable with that at the start of the experiment. There was a loss of epithelial glycogen within the first day, with occasional reappearance of patchy and irregular deposits. Whereas the control of epithelial thickness appeared to be restored after 10 days in vitro, disturbances in the maturation of the keratinocytes, manifested as epithelial pearls and dyskeratotic cells, were evident at subsequent stages. Epiboly never occurred. The connective tissue component showed continued development, indicated by an increase in the thickness of collagen fibres. The overall palatal growth seen in vivo did not occur in organ culture. We suggest that the improved maintenance reported is partly the result of explanting tissues in such a way as to minimize trauma, and partly the result of incorporating serum albumin into the chemically defined medium. (+info)
Cognitive function and treatment of obstructive sleep apnea syndrome.
Among patients with obstructive sleep apnea syndrome (OSAS), impairment of cognitive function, i.e. deficits in memory, attention, and visuconstructive abilities are common. We applied different forms of treatment for patients with newly diagnosed OSAS in a randomized study with a one-year follow-up. Patients with BMI > 40 kg/m2 were excluded. After the initial diagnostic work-up, male patients were considered to be candidates for either nasal continuous airway pressure (nCPAP) (27 patients) or surgical treatment (uvulopalatopharyngoplasty with or without mandibular osteotomy) (23 patients). Within the groups, the patients were then randomized to active treatment (nCPAP/surgery) or to conservative management. Cognitive function and severity of OSAS were assessed prior to treatment and 3 and 12 months later. At 12 months, all patients on nCPAP had a normal ODI4 index (< 10), and were significantly less somnolent than their controls; 3/11 of the surgically treated patients had a normal ODI4 index. Daytime somnolence was significantly less severe in the surgically treated patients than in their controls. Cognitive function did not correlate importantly with daytime sleepiness or severity of OSAS; the best Pearson pairwise correlation coefficient was between ODI4 and the Bourdon-Wiersma (r = 0.36). Success in treatment of OSAS did not affect neuropsychological outcome. We concluded that the standard cognitive test battery is insufficiently sensitive to identify positive changes in patients with OSAS, especially among those with a high level of overall mental functioning. (+info)
Ectopic eruption of the maxillary canine quantified in three dimensions on cephalometric radiographs between the ages of 5 and 15 years.
The eruption paths of 20 ectopic maxillary canine teeth (10 right, 10 left) were measured in three dimensions on annual lateral and depressed postero-anterior cephalometric radiographs of 15 patients between the ages of 5 and 15 years and compared with the eruption of normal canines. It was found that between the ages of 8 and 12 years ectopic canines on the left side moved more anteriorly than the normally erupting canines and the same was true of the right canines between the ages of 7 and 12 years. While the ectopic canines moved occlusally, their vertical movement was less than normal which accounts for the clinical finding that canines are impacted in the palate at a high level. The average palatally ectopic canine always moves palatally, and never shares in the buccal movement shown by normally erupting canines between the ages of 10 and 12 years. It was interesting to find that the differences between growth of normal and ectopic canines in the lateral plane of space are present as early as 5-6 years. (+info)
Palatal bone support for orthodontic implant anchorage--a clinical and radiological study.
When maximal anchorage is required during orthodontic treatment, additional aids are often needed to support the anchoring teeth. While intra-oral aids may be limited in their anchorage potential, extra-oral anchoring aids are often rejected by the patients. Endosseous implants may therefore be a valuable alternative for stable intra-oral anchorage. However, the possibility of using conventional implants is insufficient, e.g. for treating purely orthodontic patients with full dentition or where extraction sites are to be closed. Therefore, the mid-sagittal area of the palate is an alternative insertion site for the placement of implants for orthodontic anchorage. The limited bone height in this area inspired this comparison between bone thickness in the implantation site as verified by probing during the implantation of Straumann Ortho-system implants, and thickness as measured on the lateral cephalogram. The results suggest that vertical bone support is at least 2 mm higher than apparent on the cephalogram. In none of 12 patients was a perforation to the nasal cavity found. However, in five subjects the implant projected into the nasal cavity on the post-operative cephalogram. These results were supported by the study of the projections of palate and wires in wire-marked skulls where the wires were placed bilaterally on the nasal floor and on the nasal crest. It is therefore concluded that the mid-sagittal area of the palate lends sufficient bony support for the implantation of small implants (4-6 mm endosseous length, diameter 3.3 mm). (+info)
The robust australopithecine face: a morphogenetic perspective.
The robust australopithecines were a side branch of human evolution. They share a number of unique craniodental features that suggest their monophyletic origin. However, virtually all of these traits appear to reflect a singular pattern of nasomaxillary modeling derived from their unusual dental proportions. Therefore, recent cladistic analyses have not resolved the phylogenetic history of these early hominids. Efforts to increase cladistic resolution by defining traits at greater levels of anatomical detail have instead introduced substantial phyletic error. (+info)