The etiology of cleft palate formation in BMP7-deficient mice

Palatogenesis is a complex process implying growth, elevation and fusion of the two lateral palatal shelves during embryogenesis. This process is tightly controlled by genetic and mechanistic cues that also coordinate the growth of other orofacial structures. Failure at any of these steps can result in cleft palate, which is a frequent craniofacial malformation in humans. To understand the etiology of cleft palate linked to the BMP signaling pathway, we studied palatogenesis in Bmp7-deficient mouse embryos. Bmp7 expression was found in several orofacial structures including the edges of the palatal shelves prior and during their fusion. Bmp7 deletion resulted in a general alteration of oral cavity morphology, unpaired palatal shelf elevation, delayed shelf approximation, and subsequent lack of fusion. Cell proliferation and expression of specific genes involved in palatogenesis were not altered in Bmp7-deficient embryos. Conditional ablation of Bmp7 with Keratin14-Cre or Wnt1-Cre revealed that neither epithelial nor neural crest-specific loss of Bmp7 alone could recapitulate the cleft palate phenotype. Palatal shelves from mutant embryos were able to fuse when cultured in vitro as isolated shelves in proximity, but not when cultured as whole upper jaw explants. Thus, deformations in the oral cavity of Bmp7-deficient embryos such as the shorter and wider mandible were not solely responsible for cleft palate formation. These findings indicate a requirement for Bmp7 for the coordination of both developmental and mechanistic aspects of palatogenesis.

Class II treatment by extraction of maxillary first molars or Herbst appliance: dentoskeletal and soft tissue effects in comparison

AIM To compare dentoskeletal and soft tissue treatment effects of two alternative Class II division 1 treatment modalities (maxillary first permanent molar extraction versus Herbst appliance). METHODS One-hundred-fifty-four Class II division 1 patients that had either been treated with extractions of the upper first molars and a lightwire multibracket (MB) appliance (n = 79; 38 girls, 41 boys) or non-extraction by means of a Herbst-MB appliance (n = 75; 35 girls, 40 boys). The groups were matched on age and sex. The average age at the start of treatment was 12.7 years for the extraction and for 13.0 years for the Herbst group. Pretreatment (T1) and posttreatment (T2) lateral cephalograms were retrospectively analyzed using a standard cephalometric analysis and the sagittal occlusal analysis according to Pancherz. RESULTS The SNA decrease was 1.10° (p = 0.001) more pronounced in the extraction group, the SNB angle increased 1.49° more in the Herbst group (p = 0.000). In the extraction group, a decrease in SNB angle (0.49°) was observed. The soft tissue profile convexity (N-Sn-Pog) decreased in both groups, which was 0.78° more (n. s.) pronounced in the Herbst group. The nasolabial angle increased significantly more (+ 2.33°, p = 0.025) in the extraction group. The mechanism of overjet correction in the extraction group was predominantly dental (65% dental and 35% skeletal changes), while in the Herbst group it was predominantly skeletal (58% skeletal and 42% dental changes) in origin. CONCLUSION Both treatment methods were successful and led to a correction of the Class II division 1 malocclusion. Whereas for upper first molar extraction treatment more dental and maxillary effects can be expected, in case of Herbst treatment skeletal and mandibular effects prevail.

Anatomical characteristics and dimensions of edentulous sites in the posterior maxillae of patients referred for implant therapy

The purpose of this study was to analyze the width and height of edentulous sites in the posterior maxilla using cone beam computed tomography (CBCT) images from patients referred for implant therapy. A total of 122 CBCT scans were included in the analysis, resulting in a sample size of 252 edentulous sites. The orofacial crest width was measured in coronal slices, perpendicular to the alveolar ridge. The bone height was analyzed in the respective sagittal slices. Additionally, the following secondary outcome parameters were evaluated: the morphology of the sinus floor, the presence of septa in the maxillary sinus, and the thickness of the sinus membrane. The mean crest width for all analyzed sites was 8.28 mm, and the mean bone height was 7.22 mm. The percentage of patients with a crest width of less than 6 mm was 27% in premolar sites and 7.8% in molar sites. The bone height decreased from premolar to molar areas, with a high percentage of first and second molar sites exhibiting a bone height of less than 5 mm (54.12% and 44.64%, respectively). Regarding the morphology of the sinus floor, 53% of the edentulous sites exhibited a flat configuration. A septum was present in 67 edentulous sites (26.59%). Analysis of the sinus membrane revealed 88 sites (34.9%) with increased mucosal thickness (> 2 mm). For the crest width, the location of the edentulous site and the morphology of the sinus floor were both statistically significant variables. For the crest width and mean bone height, the location of the edentulous site and the morphology of the sinus floor were both statistically significant variables. The study confirmed that a high percentage of edentulous sites in the posterior maxilla do require sinus floor elevation to allow the placement of dental implants. Therefore, a detailed three-dimensional radiograph using CBCT is indicated in most patients for proper treatment planning.

Gene Discovery in Nonsyndromic Cleft Lip with or without Cleft Palate

Nonsyndromic cleft lip with or without cleft palate (NSCLP), a common, complex orofacial birth defect that affects approximately 4,000 newborns each year in the United States, is caused by both genetic and environmental factors. Orofacial clefts affect the mouth and nose, causing severe deformity of the face, which require medical, dental and speech therapies. Despite having substantial genetic liability, less than 25% of the genetic contribute to NSCLP has been identified. The studies described in this thesis were performed to identify genes that contribute to NSCLP and to demonstrate the role of these genes in normal craniofacial development. Using genome scan and candidate gene approaches, novel associations with NSCLP were identified. These include MYH9 (7 SNPs, 0.009≤p<0.05), Wnt3A (4 SNPs, 0.001≤p≤0.005), Wnt11 (2 SNPs, 0.001≤p≤0.01) and CRISPLD2 (4 SNPs, 0.001≤p<0.05). The most interesting findings were for CRISPLD2. This gene is expressed in the fused mouse palate at E17.5. In zebrafish, crispld2 localized to the craniofacial region by one day post fertilization. Morpholino knockdown of crispld2 resulted in a lower survival rates and altered neural crest cell (NCC) clustering. Because NCCs form the tissues that populate the craniofacies, this NCC abnormality resulted in cartilage abnormalities of the jaw including fewer ceratobranchial cartilages forming the lower jaw (three pairs compared to five) and broader craniofacies compared to wild-type zebrafish. These findings suggest that the CRISPLD2 gene plays an important role in normal craniofacial development and perturbation of this gene in humans contributes to orofacial clefting. Overall, these results are important because they contribute to our understanding of normal craniofacial development and orofacial clefting etiology, information that can be used to develop better methods to diagnose, counsel and potentially treat NSCLP patients.

Mutation and association analysis of the PVR and PVRL2 genes in patients with non-syndromic cleft lip and palate.

Orofacial clefts (OFC; MIM 119530) are among the most common major birth defects. Here, we carried out mutation screening of the PVR and PVRL2 genes, which are both located at an OFC linkage region at 19q13 (OFC3) and are closely related to PVRL1, which has been associated with both syndromic and non-syndromic cleft lip and palate (nsCLP). We screened a total of 73 nsCLP patients and 105 non-cleft controls from the USA for variants in PVR and PVRL2, including all exons and encompassing all isoforms. We identified four variants in PVR and five in PVRL2. One non-synonymous PVR variant, A67T, was more frequent among nsCLP patients than among normal controls, but this difference did not achieve statistical significance.

Mutation analysis of the PVRL1 gene in caucasians with nonsyndromic cleft lip/palate.

Nonsyndromic cleft lip with or without cleft palate (nsCL/P, MIM 119530) is perhaps the most common major birth defect. Homozygous PVRL1 loss-of-function mutations result in an autosomal recessive CL/P syndrome, CLPED1, and a PVRL1 nonsense mutation is associated with sporadic nsCL/P in Northern Venezuela. To address the more general role of PVRL1 variation in risk of nsCL/P, we carried out mutation analysis of PVRL1 in North American and Australian nsCL/P cases and population-matched controls. We identified a total of 15 variants, 5 of which were seen in both populations and 1 of which, an in-frame insertion at Glu442, was more frequent in patients than in controls in both populations, though the difference was not statistically significant. Another variant, which is specific to the PVRL1 beta (HIgR) isoform, S447L, was marginally associated with nsCL/P in North American Caucasian patients, but not in Australian patients, and overall variants that affect the beta-isoform were significantly more frequent among North American patients. One Australian patient had a splice junction mutation of PVRL1. Our results suggest that PVRL1 may play a minor role in susceptibility to the occurrence of nsCL/P in some Caucasian populations, and that variation involving the beta (HIgR) isoform might have particular importance for risk of orofacial clefts. Nevertheless, these results underscore the need for studies that involve very large numbers when assessing the possible role of rare variants in risk of complex traits such as nsCL/P.

Incidental findings on cone beam computed tomography scans in cleft lip and palate patients.

OBJECTIVES Cone beam computed tomography (CBCT) is frequently used in treatment planning for alveolar bone grafting (ABG) and orthognathic surgery in patients with cleft lip and palate (CLP). CBCT images may depict coincident findings. The aim of this study was to assess the prevalence of incidental findings on CBCT scans in CLP patients. SUBJECTS AND METHODS Initial CBCTs taken from consecutive patients (n = 187; mean age 11.7 years, range 6.9-45) with a non-syndromic orofacial cleft from January 2006 until June 2012 were systematically evaluated. Twenty-eight patients (mean age 19.3 years, range 13.2-30.9) had been subjected to ABG before their first CBCT was taken; 61 patients had a CBCT before and after ABG. Sinuses, nasopharynx, oropharynx, throat, skull, vertebrae, temporomandibular joint (TMJ), maxilla and mandible were checked for incidental findings. RESULTS On 95.1 % of the CBCTs, incidental findings were found. The most prevalent were airway/sinus findings (56.1 %), followed by dental problems, e.g. missing teeth (52 %), nasal septum deviation (34 %), middle ear and mastoid opacification, suggestive for otitis media (10 %) and (chronic) mastoiditis (9 %), abnormal TMJ anatomy (4.9 %) and abnormal vertebral anatomy (1.6 %). In the 28 patients whose first CBCT was taken at least 2 years after ABG, bone was still present in the reconstructed cleft area except in 2 out of 12 patients with a bilateral CLP. The ABG donor site (all bone grafts were taken from the chin area) was still recognizable in over 50 % of the patients. Based on the CBCT findings, 10 % of the patients were referred for further diagnosis and 9 % for further treatment related to dental problems. CONCLUSION Incidental findings are common on CBCTs. Compared with the literature, CLP patients have more dental, nasal and ear problems. Thus, whenever a CBCT is available, this scan should be reviewed by all specialists in the CLP team focusing on their specific background knowledge concerning symptoms and treatment of these patients. CLINICAL RELEVANCE The high number of findings indicates that CBCT imaging is a helpful tool in the treatment of CLP patients not only related to alveolar bone grafting and orthognathic surgery but it also provides diagnostic information for almost all specialties involved in CLP treatment.

Factors influencing fixed retention practices in German-speaking Switzerland : A survey.

AIM Several surveys evaluate different retention approaches among orthodontists, but none exist for general dentists. The primary aim of this survey was to record the preferred fixed retainer designs and retention protocols amongst general dentists and orthodontists in Switzerland. A secondary aim was to investigate whether retention patterns were associated with parameters such as gender, university of graduation, time in practice, and specialist status. METHODS An anonymized questionnaire was distributed to general dentists (n = 401) and orthodontists (n = 398) practicing in the German-speaking part of Switzerland. A total of 768 questionnaires could be delivered, 562 (73.2 %) were returned and evaluated. Descriptive statistics were performed and responses to questions of interest were converted to binary outcomes and analyzed using multiple logistic regression. Any associations between the answers and gender, university of graduation (Swiss or foreign), years in practice, and specialist status (orthodontist/general dentist) were assessed. RESULTS Almost all responding orthodontists (98.0 %) and nearly a third of general dentists (29.6 %) reported bonding fixed retainers regularly. The answers were not associated with the practitioner's gender. The university of graduation and number of years in practice had a moderate impact on the responses. The answers were mostly influenced by specialist status. CONCLUSION Graduation school, years in practice, and specialist status influence retention protocol, and evidence-based guidelines for fixed retention should be issued to minimize these effects. Based on the observation that bonding and maintenance of retainers are also performed by general dentists, these guidelines should be taught in dental school and not during post-graduate training.

Nasolabial symmetry and esthetics in cleft lip and palate: analysis of 3D facial images

OBJECTIVES To determine the relationship between nasolabial symmetry and esthetics in subjects with orofacial clefts. MATERIAL AND METHODS Eighty-four subjects (mean age 10 years, standard deviation 1.5) with various types of nonsyndromic clefts were included: 11 had unilateral cleft lip (UCL); 30 had unilateral cleft lip and alveolus (UCLA); and 43 had unilateral cleft lip, alveolus, and palate (UCLAP). A 3D stereophotogrammetric image of the face was taken for each subject. Symmetry and esthetics were evaluated on cropped 3D facial images. The degree of asymmetry of the nasolabial area was calculated based on all 3D data points using a surface registration algorithm. Esthetic ratings of various elements of nasal morphology were performed by eight lay raters on a 100 mm visual analog scale. Statistical analysis included ANOVA tests and regression models. RESULTS Nasolabial asymmetry increased with growing severity of the cleft (p = 0.029). Overall, nasolabial appearance was affected by nasolabial asymmetry; subjects with more nasolabial asymmetry were judged as having a less esthetically pleasing nasolabial area (p < 0.001). However, the relationship between nasolabial symmetry and esthetics was relatively weak in subjects with UCLAP, in whom only vermilion border esthetics was associated with asymmetry. CONCLUSIONS Nasolabial symmetry assessed with 3D facial imaging can be used as an objective measure of treatment outcome in subjects with less severe cleft deformity. In subjects with more severe cleft types, other factors may play a decisive role. CLINICAL SIGNIFICANCE Assessment of nasolabial symmetry is a useful measure of treatment success in less severe cleft types.

Are claims made in orthodontic journal advertisements evidence-supported?

OBJECTIVE To examine the supporting evidence of advertisements published in six leading orthodontic journals. MATERIALS AND METHODS The 2012-2013 printed issues of American Journal of Orthodontics and Dentofacial Orthopedics, Australian Orthodontic Journal, Journal of Orthodontics, European Journal of Orthodontics, Journal of Clinical Orthodontics, and Journal of Orofacial Orthopedics were screened for advertisements implying superior performance compared with competitor products. Advertisements were classified according to type of product, availability, and currency of supporting references. RESULTS A total of 99 unique advertisements claiming clinical benefit or superiority were identified. The overwhelming majority of the identified advertisements promoted appliance products (62.6%), orthodontic materials (14.1%), and dental operatory equipment, including imaging systems (12.1%). Advertisements were found to provide references or not regardless of the product type. Half of the advertisements referred to at least one peer-reviewed publication, whereas unpublished studies were cited by 25% of the advertisements. Most of the referenced articles were published within the past 5 years. CONCLUSIONS The scientific background of advertisements in the orthodontic literature appears limited. While surveillance of journal advertising needs to be regulated, clinicians are urged to critically appraise the claims being made in orthodontic print advertisements by consulting the associated existing evidence.

Catchment area of oral cleft cases for two major hospitals in Harris County

Background. Orofacial clefts are among the most common birth defects and considered to be of complex etiology with both genetic and environmental factors.^ Objectives. The purpose of this study was to describe maternal and infant characteristics, examine the catchment area, and determine if there are any geospatial patterns among infants with an orofacial cleft delivered at two major hospitals in Harris County, The Woman's Hospital of Texas and Memorial Hermann Hospital-Texas Medical Center, from January 1, 2003 through December 31, 2007.^ Methods. Data were obtained from two major hospitals in Harris County and included all babies delivered in the period from 2003 through 2007 with an orofacial cleft. Residential addresses were mapped using MapInfo GIS software and the cluster analysis performed with SaTScan software.^ Results. Ninety-nine cases were identified spanning nine counties. 26% of cases resided within a 5-mile radius of the Texas Medical Center. Birth rates ranged from 1.4 to 16.5 per 10,000 total births. A cluster was identified in southwest Harris County, however, it was not significant (p=0.066).^ Conclusion. This study encourages further focus on linking cleft cases to environmental factors in order to determine potential risks. ^

Conservación de zapallos (Cucurbita pepo L.) var. Frontera : cultivo orgánico y tradicional

Objetivos: Determinar el período de conservación de zapallo orgánico respecto del tradicional, en dos condiciones de almacenamiento. Materiales y métodos: Zapallos variedad Frontera se limpiaron en seco y se pesaron individualmente para controlar pérdida de peso; se consignaron características externas. Para determinar deshidratación se midió humedad en la pulpa (estufa 105 °C). Para conservación, acondicionamiento en cajones de madera con viruta, en galpón (temperatura y humedad ambiente) y antecámara (15- 20 °C y 70-85 % HR). Duración 187 días. Comparación sensorial, al inicio y al final de la conservación, considerando: color, olor, sabor, retrogusto, jugosidad, textura y calidad general. Se utilizó planilla con escala mixta estructurada de 9 puntos y participaron jueces semientrenados (n = 10). Evaluación de datos por análisis de varianza = 0.05. Conclusiones: En las condiciones del ensayo, el período de conservación no debe ser superior a los 105 días por aparición de podredumbre. Por deshidratación superior al 60 %, debe ser inferior a 68 días para antecámara, 105 días para orgánicos de galpón y 105-126 días para tradicionales de galpón. Las características sensoriales iniciales de los zapallos orgánicos fueron mejores; al final se destacaron los tradicionales en galpón, luego los orgánicos en galpón y los conservados en antecámara presentaron características inferiores.

Aceite esencial de tomillo como antioxidante y conservador de hamburguesas funcionales

Para determinar el poder antioxidante y conservante del aceite esencial de tomillo mendocino (Acantholippia seriphioides) en hamburguesas funcionales, conservadas a 4 ± 0.5 °C, se elaboraron medallones utilizando: 83 % carne vacuna 5 % grasa vacuna 5 % salvado de avena 5 % texturizado de soja 2 % sustituto graso 0.08 g NaCl/kg aceite esencial de tomillo (AET) 106 ufc/g Se estudió: • tipo de envasado: bolsas de poliamidapolietileno; atmósfera modificada: 70 % N 2 y 30 % CO2, y vacío; • tiempo de almacenamiento: 0, 1, 2, 3 y 4 semanas; • dosis de tomillo: 0 y 150 mg AET/kg Los datos se analizaron de acuerdo con un arreglo factorial 2 (tipo de envasado) x 5 (tiempos de almacenamiento) x 2 (dosis de AET) en un diseño de parcelas subdivididas, con 4 repeticiones. Se evaluaron TBA, pH, NBV, color, olor y carga microbiana. Las tres primeras variables se estudiaron mediante el ANOVA. Para color y olor se recurrió a un análisis sensorial descriptivo. La carga microbiana se representó gráficamente. El AET disminuyó el TBA, independientemente del tipo de envasado utilizado. El pH disminuyó en el tiempo siguiendo un modelo polinomial de 2° grado. El NBV resultó significativo para: envasamiento al vacío y 0 mg/kg AET. El AET mantiene los atributos sensoriales durante las dos primeras semanas pero no disminuye la carga microbiana. Conclusión: el AET tiene efecto antioxidante pero no conservante en hamburguesas funcionales de carne vacuna bajo las condiciones del ensayo.

Evaluación sensorial de mermeladas de tomate de color no tradicional

Nuevas cultivares de tomate, de colores distintos al tradicional rojo, se adaptan a la elaboración de productos alternativos, como las confituras. Se estudió la aceptabilidad por parte del consumidor de mermeladas elaboradas con las variedades Victoria FCA, Don Armando FCA y Santa Rosa FCA. Sus frutos: amarillos, anaranjados y rojos, respectivamente, fueron caracterizados por color, peso, acidez: titulable y potencial, y sólidos solubles. Las mermeladas, aromatizadas con clavo de olor, se elaboraron en una planta experimental hasta concentración 67-69 % de sólidos solubles. Un panel de 39 consumidores -clasificados en menores y mayores de 30 años- evaluó aspecto, color, aroma, textura y sabor, aplicando escalas no estructuradas. Las evaluaciones de ambos grupos fueron distintas. Para todas las características sensoriales la prueba de Friedman indicó diferencias entre los tres productos (a = 0,001). En una escala para cinco categorías, más del 50 % de los jueces consideraron las tres mermeladas en las categorías más altas: me gusta y me gusta mucho. El análisis de los datos categóricos de preferencia otorgó el primer lugar a la variedad roja, seguida por la anaranjada y la amarilla. Podría existir un segmento de consumidores interesados en el desarrollo de confituras de tomate amarillo, pero en el caso específico de la mermelada, tuvo mayor aceptabilidad el producto de color igual o parecido al tradicional.

Evaluación sensorial del impacto ambiental : industrias vitivinícola y aceitera

El objetivo de este trabajo fue probar y poner a punto un método para la valoración del impacto sobre el ambiente, generado por las industrias vitivinícolas y aceiteras. Sus efluentes líquidos se evaluaron sensorialmente durante dos años, en momentos y horarios clave de producción. Se practicaron pruebas de valoración de intensidad -mediante escalas de intervalo- y de ordenamiento, tanto para atributos: olor, color, turbidez y sedimento de efluentes líquidos, como para impactos: visual, olfativo y auditivo. Se caracterizaron las variables en cada tipo de establecimiento y por etapa considerada. Se elaboró una cartilla de color de efluentes líquidos mediante comparación con la escala Munsell. Se obtuvieron perfiles sensoriales de cada punto de muestreo. Las pruebas de ordenamiento (nivel de significancia = 0,05) arrojaron diferencias significativas para las variables y puntos de muestreo en estudio. Las pruebas de comparación múltiple indicaron los puntos de muestreo que debían mantenerse.