As fissuras lábio palatinas (FLP) são as malformações faciais mais comuns presentes ao nascimento e correspondem a cerca de 25% de todos os defeitos congênitos. Sua incidência é alta, de aproximadamente 1/700 nascimentos e varia entre os diferentes grupos étnicos. Elas representam um problema relevante de saúde, pois os portadores necessitam para sua reabilitação estética e funcional de uma equipe multidisciplinar, do nascimento e geralmente, até a vida adulta, o que exige um alto custo econômico. A etiologia das FLP não sindrômicas ainda é controversa na literatura, apesar de existirem esforços mundiais para identificar os fatores que causam esta patologia. Nosso trabalho consistiu do estabelecimento e caracterização de 23 linhagens de células tronco adultas (CTA) obtidas de polpa de dentes decíduos (11 indivíduos, sendo 5 controles, 5 portadores de FLP não sindrômica e 1 portador da síndrome de Van der Woude), do músculo orbicular do lábio (5 indivíduos portadores de FLP), do osso medular (2 indivíduos portadores de FLP) e de gordura de lipoaspiração (5 controles), com o objetivo de utilizá-las para ajudar a elucidar os mecanismos etiológicos de origem genética das fissuras lábio palatinas não sindrômicas; bem como para utilizá-las para bioengenharia dos tecidos alterados nesta patologia, especialmente o tecido ósseo. Demonstramos que os protocolos que utilizamos são facilmente reproduzidos e que as diferentes linhagens CTA obtidas tem plasticidade para diferenciação osteogênica “in vitro”, e que as CTA obtidas de polpa de dente e de músculo orbicular do lábio têm plasticidade para se diferenciar “in vitro” em tecido ósseo, muscular, condrogênico e adipogênico. Além disso, em nossos experimentos, demonstramos que estas CTA tem capacidade de regenerar grandes defeitos ósseos em ratos Wistar não imunossuprimidos, nos quais não observamos clinicamente nenhum tipo de rejeição destas células humanas. Utilizamos uma metodologia inédita para estudar a etiopatogenia das FLP, que consistiu da obtenção de RNA das CTA de indivíduos portadores de FLP e de controles para posterior hibridação em lâminas de “microarray” e comparação do perfil de expressão gênica das mesmas. Observamos neste estudo preliminar que 367 genes estão diferencialmente expressos nas amostras, sendo todos mais expressos nas amostras dos portadores de FLP e sugerimos que esta é uma estratégia promissora para estudo da etiopatogenia das FLP. Concluimos que a realização de estudos das CTA de afetados por FLP e de controles são importantes não só para utilização das mesmas em bioengenharia de tecidos, mas também para estudo da etiopatogenia das fissuras. / Clefts involving the lip and palate (CLP) or palate only (CP) represent the most frequent facial malformation during birth and correspond to approximately 25% of all congenital defects. The incidence of this problem is high; corresponding to approximately 1:700 births, but is different among the ethnic groups. Clefts represent a major health problem due to the requirement of a mutidisciplinary team to deal with the esthetical and funcional reabilitation needs of these patients from birth to, in some cases, adult life, resulting in a high treatment cost. There still many controversies about the ethiology of non-syndomic CLP, even though there are researchers all over the world trying to identify the factors causing this pathology. Our work consisted to establish and characterize 23 adult stem cells (ASC) linages obtained from falling teeth dental pulp (5 controls, 5 non-syndomic CLP patients and 1 Van der Woude syndrome patient), orbicular oris muscle (5 non-syndomic CLP patients), bone marrow (2 non-syndomic CLP patients) and adipose tissue (5 controls). The objectives were to use these lineages to help elucidate the genetic mechanisms involved in the development of non-syndomic CLP, as well as, to bioengineer tissues affected by this pathology, mainly bone tissue. The protocols we used are easily reproductible and we demonstrate that our different ASC linages have in vitro plasticity to osteogenic differentiation, as well as, the lineages obtained from dental pulp and orbicular oris muscle have osteogenic, muscular, condrogenic and adipogenic plasticity. Further more our experiments demonstrate that these ASC lineages have the capacity to regenerate major bone defects in non-immunocompromised Wistar rats, which did not develop any kind of rejection against these human cells. We have used a new methodology to study the molecular mechanisms involved in CLP, which consisted in comparing the mRNA expression profile of ASC from non-syndomic CLP patients and controls, using microarrays. We observed that 367 genes were differently expressed between the two groups and all these genes have a higher expression in samples from nonsyndomic CLP patients, thus suggesting that this is a promising strategy to study the molecular mechanisms involved in CLP. In conclusion, studies involving ASC from non-syndomic CLP patients and controls are important not only to tissue bioengineering, but also to elucidate the molecular mechanisms involved in CLP.

A fissura lábio-palatina não-sindrômica (FLP NS) é uma doença multifatorial, determinada pela interação entre fatores genéticos e ambientais e sua incidência é estimada entre 0,34 e 2,29 a cada 1000 nascimentos. Trata-se de uma embriopatia causada por erros durante a morfogênese orofacial, a qual depende de uma fina regulação de mecanismos como proliferação celular, remodelagem de matriz extracelular, e transição epitélio-mesenquimal. Apesar de intensos esforços para se determinar fatores genéticos e ambientais de susceptibilidade, a etiologia desta malformação permanece pouco compreendida. Vários loci associados às FLP NS vêm sendo identificados por meio de estudos de mapeamento gênico convencionais, entretanto, a grande maioria dos resultados não se replica em diferentes estudos, e não há clareza acerca do efeito funcional das variantes detectadas. Neste contexto, uma abordagem interessante para investigar a etiologia da doença é a análise de expressão gênica, que pode ser utilizada para identificar alterações de vias biológicas que convergem na manifestação do quadro clínico. Em vista disso, neste trabalho nós utilizamos a análise do transcriptoma de células-tronco de polpa dental de pacientes portadores de FLP NS, com o intuito de identificar padrões de expressão relacionados a mecanismos biológicos relevantes para a embriopatogênese da doença. Obtivemos padrões de expressão que sugerem desregulação de mecanismos associados à remodelagem de matriz extracelular e à transição epitélio-mesenquimal. Além disso, ao utilizarmos diferentes condições de cultura celular, verificamos em uma nova amostra de pacientes a desregulação de vias biológicas relacionadas ao reparo de DNA e checkpoint do ciclo celular. Nossos dados revelam a aplicabilidade das células-tronco de polpa dental para este tipo de abordagem, e indicam que tais perfis de expressão podem levar ao acometimento da morfogênese lábio-palatina. Além disso, mostramos pela primeira vez uma conexão entre desregulação de expressão gênica e a documentada maior incidência de formas esporádicas de câncer em famílias segregando a FLP NS. Nossos resultados abrem novas possibilidades para a investigação da etiologia das FLP NS, e ajudarão na compreensão dos eventos embrionários que predispõem a essa malformação. / Non-syndromic cleft lip and palate (NSCL/P) is a multifactorial disease determined by the interplay between genetic and environmental factors, with a variable incidence of 0.34-2.29:1000 births. This malformation arises from errors during lip and palate morphogenesis, which requires tight regulation of biological mechanisms such as cellular proliferation, extracellular matrix remodelling, and epithelial-mesenchymal transition. Albeit much effort has been put into determining the genetic and environmental factors underlying disease susceptibility, the aetiology of NSCL/P remains obscure. Many candidate loci have been identified through conventional gene mapping strategies, however, there is a general lack of reproducibility across studies, and there is no consensus with regard to the functional implications of the identified genetic variants. In this context, an alternative approach resides in assessing differential expression patterns to identify alterations in biological networks that could lead to phenotype manifestation. Here, we analysed the transcriptome of dental pulp stem cells from NSCL/P patients in order to pinpoint dysregulated pathways involved in the embryopathogenesis of the disease. We encountered expression patterns related to dysregulation of extracellular matrix remodelling and epithelial mesenchymal transition. Moreover, by subjecting a novel NSCL/P sample to differential cell culture conditions, we observed abnormal transcription of genes partaking in DNA repair and cell cycle checkpoint pathways. Our results show the applicability of dental pulp stem cells to this strategy and suggest that the observed expression patterns could lead to impairment of lip and palate morphogenesis. Moreover, we described for the first time a connection between abnormal gene expression in these individuals and the elevated occurrence of sporadic cancer types in NSCL/P families. Our results open new possibilities to investigate the aetiology of NSCL/P and provide further insight into the ontogenetic events underlying disease predisposition.

O objetivo deste trabalho foi investigar a ocorrência de mutações na sequência de nucleotídeos do gene AMELX, candidato a defeitos na formação do esmalte dentário, em indivíduos com e sem fissura labiopalatina. Para análise do gene proposto, foi coletado saliva de 165 indivíduos, que foram divididos em 4 grupos: Grupo 1 - composto por 46 indivíduos com fissura labiopalatina e malformação dentária; Grupo 2 - composto por 34 indivíduos com fissura labiopalatina e sem malformação dentária; Grupo 3 - composto por 34 indivíduos sem fissura labiopalatina e com malformação dentária e Grupo 4 - composto por 51 indivíduos sem fissura labiopalatina e malformação dentária. Foi realizada a extração do DNA genômico das amostras de saliva, seguido da PCR e sequenciamento direto. Cada mutação identificada no sequenciamento foi confirmada repetindo-se a reação de sequenciamento da fita antisenso. Após a coleta dos dados no Software SeqScape® 2.6, estes foram devidamente analisados por meio de gráficos e tabelas. Das amostras submetidas ao sequenciamento genético, obteve-se um aproveitamento de 95%, 90%, 89%, 88%, 94% e 100% destas amostras dos éxons 2, 3, 4, 5, 6 e 7, respectivamente. Dos 990 fragmentos sequenciados (seis éxons em 165 amostras de saliva), 918 fragmentos (93%) foram analisados. Detectou-se alteração na sequência de bases em 37 destes fragmentos (4%), sendo 14 no Grupo 1 (1,5%), 12 no Grupo 2 (1,3%), quatro no Grupo 3 (0,4%) e sete no Grupo 4 (0,7%), dos tipos missense e silenciosa, distribuídas nos éxons 2, 6 e 7 do gene AMELX, em oito distintos locais no cromossomo X. De acordo com os resultados obtidos do sequenciamento direto do gene AMELX, foi possível concluir que foram encontradas mutações na sequência de nucleotídeos do gene AMELX, em indivíduos com e sem fissura labiopalatina e malformação dentária. Observou-se ainda que a mutação localizada na posição 75 do éxon 6 esteve presente em todos os grupos estudados, sugerindo que, apesar de ser uma mutação silenciosa, pode ser um polimorfismo novo, a ser catalogado, pois foi detectado em 26 indivíduos, do total de 165, representando 16%. Entretanto, este estudo não pode afirmar que estas mutações alteraram diretamente o fenótipo dos pacientes dos grupos estudados. / The aim of this study was to investigate the occurrence of mutations in the AMELX candidate gene involved in enamel formation, in patients with and without cleft lip and palate and dental malformation. For gene analysis proposed was collected saliva from 165 patients who were divided in 4 groups: Group 1 - 46 individuals with cleft lip and palate and dental malformation, Group 2 - 34 individuals with cleft lip and palate without dental malformations; Group 3 - 34 individuals without cleft lip and palate with dental malformations and Group 4 - 51 individuals without cleft lip and palate and dental malformation. Next, genomic DNA was extracted from this saliva, followed by PCR and direct DNA sequencing. All samples with mutations were sequenced twice; once using the forward primer and a second time using the reverse primer. After data analysis with Software SeqScape® 2.6, the data were collated. Of the 165 samples, 95%, 90%, 89%, 88%, 94% and 100% of the samples were successfully sequenced from exons 2, 3, 4, 5, 6 and 7 respectively. Overall, of the 990 total sequenced exons (six exons in 165 samples of saliva), 918 exons (93%) were able to be completely sequenced and analyzed. Mutations were detected in 37 of the fragments (4%), more specifically, 14 in Group 1 (1.5%), 12 in Group 2 (1.3%), four in Group 3 (0.4%) and seven in Group 4 (0.7%), which included only missense and silent mutations, distributed throughout exons 2, 6 and 7 in the AMELX gene in eight different locations on chromosome X. According to the results obtained from direct sequencing of protein-coding exons of the AMELX gene, mutations were found in the nucleotide sequence of the AMELX gene in individuals with and without cleft lip and palate and dental malformation. It was also observed that the mutation in position 75 of exon 6 was present in all groups, suggesting that, though a silent mutation, may be a new polymorphism to be cataloged: it was found in 26 patients, the total of 165, representing 16%. However, this study cannot confirm that these mutations directly altered the phenotypes of the patients in the groups tested.

Social isolation is commonly reported in individuals with isolated cleft lip and palate (ICLP), and is often cited as due to secondary factors of living with a facial malformation. However, the data are mixed, and the literature is biased to self-report studies. This study aimed to go beyond self-report data, and various components of social functioning were examined in a group of males with and without ICLP. The main aim of the study was to assess and compare social motivation in ICLP, and to relate social adjustment to brain structure. It was predicted that males with ICLP would be more likely to be socially isolated; however, self-concept was predicted to be similar to the comparison group (NC). Additionally, males with ICLP were predicted to have reduced social motivation (i.e., social abulia). Finally, volumes of the medial orbitofrontal cortex (mOFC) and the cerebellum were predicted to be related to social outcomes. The sample's age range was 13 - 25 years old, and 20 males with ICLP were compared to a group of 20 NC males. MRI scans were obtained from all the participants. As expected, males with ICLP were more likely to be socially isolated. Against predictions, they also had lower self-concept relative to the NC group. However, self-concept was not related to the extent of facial abnormality in the ICLP group. In line with predictions, the study did provide evidence for social abulia as a mechanism for social isolation, as males with ICLP had a more positive attitude after being socially excluded relative to excluded NC males. Unexpectedly however, the groups responded the same to social pressure, as all participants were more likely to take riskier turns in a driving simulator experiment when someone behind them was honking. Finally, social adjustment was significantly correlated with the volume of the mOFC, and posterior cerebellum white matter. Both correlations suggested that individuals with larger volumes were more likely to be better socially adjusted. In conclusion, the study provided evidence for a potentially different mechanism of social isolation in ICLP, and showed that brain morphology may at least partly underlie social dysfunction as well.

abstract: The purpose of this study is to evaluate the effects of parent training in the Enhanced Milieu Teaching with Phonological Emphasis (EMT+PE) intervention program, using a secure internet-based conferencing software (telepractice), on parent strategy use and child speech and language outcomes for children with repaired cleft lip and/or palate (CL/P). Three participant dyads composed of a parent and child participated in this study. Children ranged in age from 21 to 27 months at the beginning of this study and all had a diagnosis of nonsyndromic CL/P. Participating dyads received three in- person training sessions and three weekly telepractice intervention sessions. Assessment and intervention sessions were administered by a trained Speech Language Pathologist (SLP) and a graduate SLP student clinician. Parents demonstrated a positive intervention effect by significantly increasing their use of EMT+PE intervention strategies during training. Based on preliminary results, parents were able to maintain their increased use of strategies following the conclusion of intervention as well. Telepractice proved to be a valid service delivery model for conducting early intervention sessions and for supporting the early speech and language development for children with CL/P. / Dissertation/Thesis / Masters Thesis Communication Disorders 2020

Cleft lip and palate (CLP) has been determined to be the second most common birth defect in the United States, affecting 1 in every 940 births (Parker et al., 2010). The team approach is the accepted best practice for children with CLP (Kummer, 2020) and the school-based Speech-Language Pathologist (SLP) has an important role to play in assessment and intervention of children with repaired CLP, however there is little research to describe their collaboration. This research aimed to explore and describe the referral practices of school-based SLP’s to CLP teams. A survey titled “Referral to Cleft Lip and Palate Teams: Practice of School-Based Speech Language Pathologist’s” was developed and distributed to members of the American Speech-Language Hearing Association’s (ASHA’s) Special Interest Groups (SIGs) 15 and 16 following an in depth literature review on the topic. A total of 57 practicing school-based SLPs acted as respondents. The results of the survey suggested VPD was the main reason for making a referral to a CLP team (89.72%), which validates the response that clients mostly referred had suspected VPD (89.47%). Making a team referral was not common practice, as 58.7% had never made a CLP team referral in the schools. ENTs (51.06%) were the preferred choice of referral in comparison to a CLP team (25.53%). Barriers to making CLP team referrals varied and obtaining permission from the school was experienced by some respondents (36.36%). Respondents made valuable comments which centered on positive experiences with working with CLP teams (11/56). The process of making referrals to CLP teams and collaboration between school-based SLPs and CLP teams needs to be addressed in graduate training and CE. According to Vallino et al., (2019) such communication enhances care, bridges the perceived gap between school-based SLPs and CLP teams, and will ensure that children with CLP and VPD receive the best care possible.

In der vorliegenden Dissertation wurde anhand von 1042 extraoralen Fotos im Profil und en face untersucht, wie sich das Wachstum longitudinal bei 166 gesunden Kindern, je zehn Kindern mit isolierter Gaumenspalte, einseitiger Lippen-Kiefer-Gaumenspalte sowie doppelseitiger Lippen-Kiefer-Gaumenspalte operiert nach dem Leipziger Konzept entwickelt, sowie inwiefern sich die einzelnen Gruppen in verschiedenen Altersstufen voneinander unterscheiden. Dazu wurden pro Gruppe jeweils fünf Jungen und fünf Mädchen kaukasischer Herkunft, geboren in den Jahren 1994 bis 2001, mit einer nichtsyndromalen Spalte ausgewählt, die ab der Geburt über mehrere Jahre hinweg fotografiert wurden. Diese Fotos wurden digitalisiert und denen von Kontrollpatienten gegenüber gestellt. Anhand von Winkeln der klassischen Morphometrie und Grafiken und Statistiken der geometrischen Morphometrie wurden die Veränderungen im Wachstum analysiert und die Kontrollpatienten mit dem Leipziger OP-Konzept verglichen. Es konnte gezeigt werden, dass die geometrische Morphometrie durchaus geeignet ist, das Gesichtswachstum anhand von extraoralen Fotos zu beurteilen. Weiterhin konnte bestätigt werden, dass mit dem Leipziger Konzept zur funktionellen Rehabilitation von Patienten mit Lippen-Kiefer-Gaumenspalten sehr gute Operationsergebnisse und vor allem Langzeitergebnisse erzielt werden können.

There has been a rise non-governmental organizations (NGOs) as part of a global health system that seeks to treat children with cleft lip and palate (CLP) in resource-poor countries. As a craniofacial abnormality, CLP affects a child’s ability to communicate and consume food, and the stigma associated with the condition leads to both social and physiological suffering. International NGOs use an apolitical humanitarian rhetoric to justify the need to provide this life-saving surgery. This thesis assesses CLP interventions by applying a critique of neoliberalism to explore the ways economic rationalities are extended to the domain of humanitarianism. By employing an ethnographic approach of “studying up,” this thesis critiques a North American NGO, referred to as Mission Smile. To reveal how neoliberal rationalities are embedded within the organization, this research draws on data from media analysis, participant observation, and interviews with medical volunteers and employees at Mission Smile. This thesis argues that neoliberal rationalities permeate throughout the organization. Economic calculus are not only embedded in the organization’s goal to provide surgery to “as many children as possible,” but also undergirds the distribution of humanitarian aid. Moreover, the surgery Mission Smile provides is described as an “investment in a child’s future” that enable children with CLP to become a contributing member of society. While this study reveals how neoliberal rationalities can converge with values of humanitarianism, it also shows that the extension of neoliberal rationalities into new domains is not a cohesive process. Volunteers describe an emergence of communitas, a feeling of bubbling joy and a shared humanity, and a development of a moral relationship with their recipients that lies partially outside the domain of market rationalities. / Graduate

December 2011. A thesis submitted to the College of Dental Medicine of Nova Southeastern University of the degree of Master of Science in Dentistry. Introduction: The nasoalveolar molding appliance is used pre-surgically in cleft lip and palate patients as a method of bringing together the lip and alveolus by applying force to direct desired growth.1 It is used in the infant to reduce the pre-surgical severity of the initial cleft and to improve alignment of the base of the nose and lip segments.1 The purpose of this study was to examine the long-term effect of nasoalveolar molding on facial growth and nasolabial esthetics in complete unilateral cleft lip and palate patients. Methods: Sixteen (N=16) post-surgical cleft lip and palate patients who had undergone nasoalveolar molding as infants and twelve (N=12) control patients, treated surgically without nasoalveolar molding, were recalled for a clinical examination including impressions, photographs, and a lateral cephalogram. Dental models were analyzed using the Goslon Yardstick, developed by Mars et al. in 1987.2¬ Photographs were analyzed using the Asher-McDade method for rating the nasolabial appearance in patients with cleft lip and palate.3 Finally, lateral cephalograms were digitized and analyzed using Dolphin Imaging software. Results: Separate ordinal logistic regression models indicated no significant difference between the molding and non-molding groups in Goslon score, nasal form, nose symmetry, vermilion border or nasolabial profile assessments. Generalized linear models revealed one cephalometric variable to be statistically significant between the two groups. The ANB angle was decreased by 2.34 degrees on average in the group who underwent nasoalveolar molding prior to cheiloplasty. Intra-rater and inter-rater weighted kappa statistics were calculated for each variable. Conclusion: Short-term benefits of nasoalveolar molding have been documented in the literature. However, more long-term studies are needed in order to demonstrate the longitudinal effects of the appliance on esthetics and midfacial growth. Due to the limitations of cleft lip and palate studies, it is often difficult to accurately assess treatment effects. Through inter-center studies such as the Eurocleft and Americleft projects, some limitations and biases can be overcome in order to compare various protocols and outcomes.4 In the future, it is the desire of the investigators to include the current sample of nasoalveolar molding patients in the future efforts and expansion of the Americleft study.

No description available.