Heleni Vastardis

Human NELL-1 expressed in unilateral coronal synostosis

Surgical correction of unilateral coronal synostosis offers a unique opportunity to examine the molecular differences between an abnormal and a normal cranial suture. We isolated and identified a cDNA fragment whose expression was up‐regulated in the premature fusing and fused coronal sutures, as compared with normal coronal sutures. The nucleotide sequence of the full‐length cDNA of this gene, human NELL‐1, has ∼61% homology with the chicken Nel gene. Both chicken Nel and human NELL‐1 are comprised of six epidermal growth factor‐like repeats. The human NELL‐1 messages were localized primarily in the mesenchymal cells and osteoblasts at the osteogenic front, along the parasutural bone margins, and within the condensing mesenchymal cells of newly formed bone in sites of premature sutural fusion. Human multiorgan tissue mRNA blot showed that NELL‐1 was specifically expressed in fetal brain but not in fetal kidney, liver, or lung. We also showed that Nell‐1 was expressed in rat calvarial osteoprogenitor cells and was largely absent in rat tibiae and fibroblast cell cultures. In conclusion, our data suggest that the NELL‐1 gene is preferentially expressed in cranial intramembranous bone and neural tissue (both of neural crest cell origin) and is up‐regulated during unilateral premature closure of the coronal suture. The precise role of this gene is unknown.

Haploinsufficiency of MSX1: a Mechanism for Selective Tooth Agenesis

ABSTRACT Previously, we found that the cause of autosomal dominant selective tooth agenesis in one family is a missense mutation resulting in an arginine-to-proline substitution in the homeodomain of MSX1. To determine whether the tooth agenesis phenotype may result from haploinsufficiency or a dominant-negative mechanism, we have performed biochemical and functional analyses of the mutant protein Msx1(R31P). We show that Msx1(R31P) has perturbed structure and reduced thermostability compared with wild-type Msx1. As a consequence, the biochemical activities of Msx1(R31P) are severely impaired, since it exhibits little or no ability to interact with DNA or other protein factors or to function in transcriptional repression. We also show that Msx1(R31P) is inactive in vivo, since it does not display the activities of wild-type Msx1 in assays of ectopic expression in the limb. Furthermore, Msx1(R31P) does not antagonize the activity of wild-type Msx1 in any of these assays. Because Msx1(R31P) appears to be inactive and does not affect the action of wild-type Msx1, we propose that the phenotype of affected individuals with selective tooth agenesis is due to haploinsufficiency.

Overexpression of Nell-1, a Craniosynostosis-Associated Gene, Induces Apoptosis in Osteoblasts During Craniofacial Development†

We studied the cellular function of Nell‐1, a craniosynostosis‐related gene, in craniofacial development. Nell‐1 modulates calvarial osteoblast differentiation and apoptosis pathways. Nell‐1 overexpression disrupts these pathways resulting in craniofacial anomalies such as premature suture closure.

Evaluation of cervical spine abnormalities on cephalometric radiographs

Cephalometric radiographs, a key element of orthodontic diagnosis, contain useful information related to the cervical spine often neglected by orthodontists and medical specialists. This article reviews cervical spine anatomy in a manner that will enable the clinician to trace the cervical spine accurately and detect cervical spine abnormalities. Examples of syndromic, nonsyndromic, and idiopathic anomalies of the cervical spine are presented and their significance discussed. Cephalometric radiographs can be used by clinicians as a potential resource for screening for pathologic abnormalities of the cervical spine and potentially averting some pathologic complications.

Signaling mechanisms implicated in cranial sutures pathophysiology: Craniosynostosis

Normal extension and skull expansion is a synchronized process that prevails along the osteogenic intersections of the cranial sutures. Cranial sutures operate as bone growth sites allowing swift bone generation at the edges of the bone fronts while they remain patent. Premature fusion of one or more cranial sutures can trigger craniosynostosis, a birth defect characterized by dramatic manifestations in appearance and functional impairment. Up until today, surgical correction is the only restorative measure for craniosynostosis associated with considerable mortality. Clinical studies have identified several genes implicated in the pathogenesis of craniosynostosis syndromes with useful insights into the underlying molecular signaling events that determine suture fate. In this review, we exploit the intracellular signal transduction pathways implicated in suture pathobiology, in an attempt to identify key signaling molecules for therapeutic targeting.

Cephalometric evaluation of the craniofacial complex in patients treated with an intraoral distraction osteogenesis device : A long-term study

INTRODUCTION Distraction osteogenesis has gained popularity because of the hypothesized concurrent soft-tissue expansion, which is believed to reduce postoperative relapse. Although many articles describe the immediate success of mandibular distraction, little research has been done on its long-term stability. Our goal was to examine the long-term craniofacial changes after distraction. METHODS Four hemifacial microsomic patients treated with unilateral mandibular distraction were recalled. Changes in maxillary width and height, occlusal height, ramus height, mandibular length, and chin position were quantified by using the posteroanterior and 45 degrees lateral oblique cephalographs. Predistraction and postdistraction measurements were taken over a 5-year period. The data were analyzed by using paired t tests and ANOVA. RESULTS Maxillary height, ramus height, mandibular length, and chin point deviation all experienced moderate improvement after distraction. Although the growth patterns between the control side and the treated side were comparable until 2 years after removal of the device, the normal side outgrew the affected side thereafter until 5 years after distraction. CONCLUSIONS Because of the greater inherent growth potential of the unaffected side, more overcorrection than originally believed is needed to offset the persistent asymmetry in growing hemifacial microsomia patients who undergo unilateral distraction osteogenesis.

Evidence for APOBEC3B mRNA and protein expression in oral squamous cell carcinomas

It has been demonstrated that APOBEC3B possesses cytidine deaminase activity, which is likely to result in C-to-T signature mutations. Increased expression of the APOBEC3B gene has been shown to correlate with higher incidence of such mutations in various cancer types, such as breast, bladder, lung, and head and neck carcinomas. In the current study, we used in silico methods, immunohistochemistry and qRT-PCR to detect the presence of APOBEC3B signature mutations and examine the levels and patterns of APOBEC3B expression in oral squamous cell carcinomas (OSCCs). Using the Cancer Genome Atlas (TCGA) database, we have found a high incidence of C-to-T transitions in head and neck squamous cell carcinomas (HNSCCs), of which OSCCs constitute the largest subgroup. Additionally, we compared APOBEC3B expression, at both mRNA and protein level, between OSCCs and non-cancerous samples. APOBEC3B was detected in both groups, but nuclear localization was consistent only in normal oral cells. APOBEC3B mRNA levels were clearly higher in OSCCs than in controls. These results suggest that while in normal oral cells APOBEC3B has an important nuclear function to fulfill, this activity may be hindered in a subgroup of tumor cells, due to the more prominent localization of the enzyme in the cytoplasm.

Unilateral Coronal Synostosis: A Histomorphometric Study

Objective This histomorphometric study compared the open and prematurely fused side of the coronal suture in subjects with unilateral coronal synostosis (UCS). Methods Sutures and parasutural bone were obtained from seven subjects with nonsyndromic UCS during operative correction at 3 to 24 months of age. Histological and cellular analyses were performed for the affected and open sutures. Specimens were examined by light and polarizing microscopy. Sutural patterns, osseous morphology, calvarial thickness, tartrate-resistant acid phosphatase (TRAP)-positive cells, and marrow spaces were evaluated histomorphologically, qualitatively, and semiquantitatively. Histomorphometry was performed to determine total projected area of marrow space as a percentage of unit area, total number of TRAP-positive cells per specimen, and perisutural cranial thickness. Results Polarizing microscopy showed that affected sutures were composed of more lamellar bone than the normal sutures. By light microscopy, the clinically fused sutures were 1.7-fold thicker (p < .02), had twofold larger marrow spaces (p < .0006), and contained sixfold more TRAP-positive osteoclasts in marrow spaces near the suture (p < .04) than the normal sutures. Quantitative analysis of the normal sutures revealed that calvarial thickness was greater with age and that there was an inverse correlation between medullary area and age. For the affected sutures, there was also an age-related increase in calvarial thickness. There were also trends for age-related declines in numbers of osteoclasts in both open and affected sides. Conclusions These results question the hypothesis that defective osteoclastic activity is pivotal in the pathogenesis of UCS and support the hypothesis that this condition results from abnormally active bony remodeling.

Saliva Proteomics Analysis Offers Insights on Type 1 Diabetes Pathology in a Pediatric Population

The composition of the salivary proteome is affected by pathological conditions. We analyzed by high resolution mass spectrometry approaches saliva samples collected from children and adolescents with type 1 diabetes and healthy controls. The list of more than 2000 high confidence protein identifications constitutes a comprehensive characterization of the salivary proteome. Patients with good glycemic regulation and healthy individuals have comparable proteomic profiles. In contrast, a significant number of differentially expressed proteins were identified in the saliva of patients with poor glycemic regulation compared to patients with good glycemic control and healthy children. These proteins are involved in biological processes relevant to diabetic pathology such as endothelial damage and inflammation. Moreover, a putative preventive therapeutic approach was identified based on bioinformatic analysis of the deregulated salivary proteins. Thus, thorough characterization of saliva proteins in diabetic pediatric patients established a connection between molecular changes and disease pathology. This proteomic and bioinformatic approach highlights the potential of salivary diagnostics in diabetes pathology and opens the way for preventive treatment of the disease.