Identification of genomic imbalances in oral clefts

Abstract Objective This article presents a clinical and cytogenomic approach that focuses on the diagnosis of syndromic oral clefts (OCs). Methods The inclusion criteria were individuals with OC presenting four or more minor signs and no major defects (non-syndromic oral clefts [NSOCs]) as well as individuals with OC presenting at least another major defect, regardless of the number of minor signs (syndromic oral clefts [SOCs]). The exclusion criteria included NSOC with less than four minor signs, SOC with known etiology, as well as atypical oral clefts. Results Of 1647 individuals with OC recorded in the Brazilian Database of Craniofacial Anomalies, 100 individuals were selected for chromosome microarray analysis (CMA). Among these, 44 individuals were clinically classified as NSOC and 56 as SOC. CMA was performed for both groups, and abnormal CMA was identified in 9%, all previously classified as SCO. The clinical and CMA data analyses showed a significant predominance of abnormal CMA in individuals classified as SOC (p = 0.0044); prematurity, weight, length, and head circumference at birth were significantly lower in the group with abnormal CMA. Besides, minor signs were significantly higher in this group (p = 0.0090). Conclusion The rigorous selection of cases indicates that the significant variables could help in early recognition of SOC. This study reinforces the importance of applying the CMA technique to establish the diagnosis of SOC. This is an important and universal issue in clinical practice for intervention, care, and genetic counseling.

Identification of genomic imbalances in oral clefts.

OBJECTIVE: This article presents a clinical and cytogenomic approach that focuses on the diagnosis of syndromic oral clefts (OCs). METHODS: The inclusion criteria were individuals with OC presenting four or more minor signs and no major defects (non-syndromic oral clefts [NSOCs]) as well as individuals with OC presenting at least another major defect, regardless of the number of minor signs (syndromic oral clefts [SOCs]). The exclusion criteria included NSOC with less than four minor signs, SOC with known etiology, as well as atypical oral clefts. RESULTS: Of 1647 individuals with OC recorded in the Brazilian Database of Craniofacial Anomalies, 100 individuals were selected for chromosome microarray analysis (CMA). Among these, 44 individuals were clinically classified as NSOC and 56 as SOC. CMA was performed for both groups, and abnormal CMA was identified in 9%, all previously classified as SCO. The clinical and CMA data analyses showed a significant predominance of abnormal CMA in individuals classified as SOC (p = 0.0044); prematurity, weight, length, and head circumference at birth were significantly lower in the group with abnormal CMA. Besides, minor signs were significantly higher in this group (p = 0.0090). CONCLUSION: The rigorous selection of cases indicates that the significant variables could help in early recognition of SOC. This study reinforces the importance of applying the CMA technique to establish the diagnosis of SOC. This is an important and universal issue in clinical practice for intervention, care, and genetic counseling.

CranFlow: An Application for Record-Taking and Management Through the Brazilian Database on Craniofacial Anomalies.

BACKGROUND: The World Health Organization has recognized the relevance of databases on craniofacial anomalies since . To date, there is no universal standard instrument/database focused on risk factors, clinical and genetic data collection, and follow-up that enables comparison between different populations and genotype-phenotype correlation. Although studies have shown that specific genes would impact outcomes, knowledge is not sufficient to subsidize cost-effectiveness strategies for diagnosis, surgical decision, and a multi-professional approach toward personalized medicine. METHODS: Based on a clinical genetic approach, a Web-based application named CranFlow-Craniofacial Anomalies: Registration, Flow, and Management has been developed. It prospectively collects clinical and genetic information for the Brazilian Database on Craniofacial Anomalies (syndromic and nonsyndromic orofacial cleft, 22q11.2 deletion syndrome, and other craniofacial related disorders). A comprehensive list of CranFlow's features is provided. RESULTS: We present preliminary results on 1546 cases already recorded and followed, which allows recognizing 10% of diagnosis changes. CONCLUSION: The identification of risk factors, consistent genetic approach associated with clinical data and follow-up result in valuable information to develop and improve personalized treatment and studies on genotype-phenotype correlation. Adoption of CranFlow in different clinical services may support comparison between populations. This application has the potential to contribute to improvements in healthcare, quality of services, clinical and surgical outcomes, and the standard of living of individuals with craniofacial anomalies. Birth Defects Research 110:72-80, 2018. © 2017 Wiley Periodicals, Inc.

Influence of sleep disorders on the behavior of individuals with autism spectrum disorder.

The aim of this study was to investigate the correlation between sleep disorders and the behavior of subjects with autism spectrum disorder (ASD) and control subjects using specific questionnaires. A small percentage (1.8%) of the control subjects had symptoms indicative of sleep-breathing disorders (SBD) and nocturnal sweating. Fifty-nine percent of the subjects with ASD had symptoms indicative of at least one sleep disorder, with SBD the most commonly reported (38%). In the control group, the symptoms of SBD were correlated with social, thought, attentional, aggression, externalizing and behavioral problems. In the ASD group, disorders of arousal (DA) were correlated with thinking problems, and disorders of excessive somnolence were correlated with thinking and behavioral problems. These results suggest that children and adolescents with ASD have a high frequency of sleep disorders, which in turn correlate with some of the behavioral traits that they already exhibit. Furthermore, sleep disturbances, when present in the typically developing children, also correlated with behavioral problems.

Clinical Features in Patients With 22q11.2 Deletion Syndrome Ascertained by Palatal Abnormalities.

OBJECTIVES: The aim of this study was to describe clinical features in subjects with palatal abnormalities and to assess the distribution of these features among those with and without 22q11.2 deletion. DESIGN: Descriptive cohort. PATIENTS: One hundred patients with palatal abnormalities and suspicion of 22q11.2 DS were included. METHODS: All patients were evaluated by a clinical geneticist, who completed a standardized clinical protocol. The 22q11.2 deletion screening was performed with fluorescence in situ hybridization using the TUPLE1 probe and multiplex ligation-dependent probe amplification using the P250-A1 kit. RESULTS: The 22q11.2 deletion was detected in 35 patients, in whom the most frequent clinical features were congenital heart disease (15/30 - 50%), developmental delay (19/35 - 54%), speech delay (20/35 - 57%), learning disabilities (27/35 - 77%), immunologic alterations (18/29 - 62%). In addition, the most common facial dysmorphisms in this group were long face (27/35 - 77%), typical nose (24/35 - 69%), and hooded eyelids (19/35 - 54%). Comparing features in patients with or without the deletion revealed significant differences (positively correlated with the deletion) for speech delay, learning disabilities, conductive hearing loss, number of dysmorphisms, long face, and hooded eyelids. Cleft lip and palate was negatively correlated with the deletion. CONCLUSIONS: The presence of speech delay, learning disabilities, conductive hearing loss, long face, and hooded eyelids should reinforce the suspicion of 22q11.2 DS in patients with palatal abnormalities and would help professionals direct clinical follow-up of these patients.

Investigation of 15q11-q13, 16p11.2 and 22q13 CNVs in autism spectrum disorder Brazilian individuals with and without epilepsy.

Copy number variations (CNVs) are an important cause of ASD and those located at 15q11-q13, 16p11.2 and 22q13 have been reported as the most frequent. These CNVs exhibit variable clinical expressivity and those at 15q11-q13 and 16p11.2 also show incomplete penetrance. In the present work, through multiplex ligation-dependent probe amplification (MLPA) analysis of 531 ethnically admixed ASD-affected Brazilian individuals, we found that the combined prevalence of the 15q11-q13, 16p11.2 and 22q13 CNVs is 2.1% (11/531). Parental origin could be determined in 8 of the affected individuals, and revealed that 4 of the CNVs represent de novo events. Based on CNV prediction analysis from genome-wide SNP arrays, the size of those CNVs ranged from 206 kb to 2.27 Mb and those at 15q11-q13 were limited to the 15q13.3 region. In addition, this analysis also revealed 6 additional CNVs in 5 out of 11 affected individuals. Finally, we observed that the combined prevalence of CNVs at 15q13.3 and 22q13 in ASD-affected individuals with epilepsy (6.4%) was higher than that in ASD-affected individuals without epilepsy (1.3%; p<0.014). Therefore, our data show that the prevalence of CNVs at 15q13.3, 16p11.2 and 22q13 in Brazilian ASD-affected individuals is comparable to that estimated for ASD-affected individuals of pure or predominant European ancestry. Also, it suggests that the likelihood of a greater number of positive MLPA results might be found for the 15q13.3 and 22q13 regions by prioritizing ASD-affected individuals with epilepsy.

Feeding Infants With Cleft Lip and/or Palate in Brazil: Suggestions to Improve Health Policy and Research.

Objective : To verify feeding resources used prior to corrective surgery among cleft babies from Brazil and to discuss suggestions to improve common feeding problems around the world. Design : Cross-sectional descriptive study conducted at eight medical centers. Participants : A total of 215 parents or guardians of cleft children. Methods : Interview based upon a prevalidated questionnaire. The chi-square test and comparison of means by analysis of variance were used; significance level adopted was 5% (P < .05). Results : Feeding guidelines were provided in the maternity unit to 53% of the families. Breast-feeding was encouraged among 80% of mothers, predominantly in the South (P = .016). However, follow-up after maternity discharge was not appropriately carried out and failure to breast-feed occurred in 78% of families. The feeding tube was used in 21%. According to families, for those who used the ordinary nipple, it was considered the best option by the majority (29%). Conclusion : Neonatal feeding in cleft babies is a global challenge. Reports about the difficulties encountered and successful experiences would be helpful to disseminate strategies and stimulate research directed at the large-scale applicability of neonatal feeding for cleft babies on public health. This study detected the need to increase professional training and emphasizes the need for public policies addressing neonatal referral to specialized care wherever possible. It also stimulates research into using an ordinary nipple as another resource for feeding cleft babies and suggests an international discussion about specific recommendations for humanized primary health care.

Defining new guidelines for screening the 22q11.2 deletion based on a clinical and dysmorphologic evaluation of 194 individuals and review of the literature.

The 22q11.2 deletion is the most frequent interstitial deletion in humans and presents a wide phenotypic spectrum, with over 180 clinical manifestations described. Distinct studies have detected frequencies of the deletion ranging from 0 % to 75 %, depending on the studied population and selection criteria adopted. Due to the lack of consensus in this matter, several studies have been conducted aiming to define which patients would be eligible for screening; however, the issue is still up for debate. In order to contribute to the delineation of possible clinical and dysmorphologic guidelines to optimize decision making in the clinical setting, 194 individuals with variable features of the 22q11.2 deletion syndromes (22q11.2DS) were evaluated. Group I, clinical suspicion of 22q11.2DS with palatal anomalies; Group II, clinical suspicion without palatal anomalies; Group III, cardiac malformations associated with the 22q11.2DS; and Group IV, juvenile-onset schizophrenia. Multiplex ligation-dependent probe amplification was used for screening the 22q11.2 deletion, which was detected in 45 patients (23.2 %), distributed as such: Group I, 35/101 (34.7 %); Group II, 4/18 (22.2 %); Group III, 6/52 (11.5 %); and Group IV, 0/23 (0 %). Clinical data were analyzed by frequency distribution and statistically. Based on the present results and on the review of the literature, we propose a set of guidelines for screening patients with distinct manifestations of the 22q11.2DS in order to maximize resources. In addition, we report the dysmorphic features which we found to be statistically correlated with the presence of the 22q11.2DS.

Estudo multicêntrico da época do diagnóstico de fendas orais / Time of diagnosis of oral clefts: a multicenter study

OBJETIVO: Determinar a época do diagnóstico de fendas orofaciais típicas em diferentes regiões brasileiras e sua influência na idade da correção cirúrgica. MÉTODO: Estudo prospectivo, descritivo e transversal realizado em centros médicos do Sudeste, Sul e Nordeste do Brasil. Fonoaudiólogos e geneticistas treinados realizaram entrevista, previamente validada, com pais de crianças afetadas. Utilizaram-se os programas Epi-Info e SPSS. Adotou-se nível de significância de 5 por cento (p < 0,05). RESULTADOS: A amostra contou com 215 entrevistas para análise: 21,9 por cento (47) aplicadas no Sudeste, 51,1 por cento (110) no Sul e 27 por cento (58) no Nordeste. A renda mensal no Sudeste foi maior (p < 0,05). A fenda labiopalatal foi encontrada em 61,4 por cento (132) dos casos, a palatal, em 20,9 por cento (45), e a labial, em 17,7 por cento (38). Em 75,3 por cento (162) dos casos, o diagnóstico ocorreu na maternidade, em 14 por cento (30), no pré-natal e, em 10,2 por cento (22), após a alta da maternidade. O Sudeste apresentou maior frequência de diagnóstico pré-natal (27,7 por cento), possivelmente relacionada ao maior poder aquisitivo e a oportunidades de investigação. Dos diagnósticos em maternidades, 74,4 por cento ocorreram no Nordeste. Entretanto, não houve diferença na comparação entre época de diagnóstico, região e idade da primeira cirurgia. CONCLUSÃO: Considerando que o diagnóstico é mais frequente em maternidades, sugere-se o treinamento das equipes de saúde desses locais, visando efetiva coordenação do atendimento inicial. Apesar da época do diagnóstico não influenciar a idade das cirurgias, ela favorece o planejamento dos cuidados neonatais e terapêuticos dos afetados.

OBJECTIVE: To determine the time of diagnosis of typical orofacial clefts in different Brazilian regions and its influence on age at surgical correction. METHOD: This was a prospective, descriptive, cross-sectional study conducted in medical centers in the Southeast, South, and Northeast of Brazil. Trained speech therapists and geneticists interviewed the parents of affected children using a previously validated questionnaire. Epi-Info and SPSS were used for data analysis. Significance level was set at 5 percent (p < 0.05). RESULTS: The sample consisted of 215 interviews conducted in the following regions: 21.9 percent (47) in the Southeast, 51.1 percent (110) in the South, and 27 percent (58) in the Northeast. Monthly family income was higher in the Southeast (p < 0.05). Cleft lip and palate were found in 61.4 percent (132) of cases, cleft palate in 20.9 percent (45), and cleft lip in 17.7 percent (38). Diagnosis occurred in the maternity ward in 75.3 percent (162) of cases, during the prenatal period in 14 percent (30), and after hospital discharge in 10.2 percent (22). The Southeast had a higher frequency of prenatal diagnosis (27.7 percent), possibly related to greater purchasing power in this region and greater availability of prenatal investigation. Of all cases diagnosed in the maternity ward, 74.4 percent occurred in the Northeast. However, no significant difference was found when comparing time of diagnosis, region, and age at first surgery. CONCLUSION: Considering that diagnosis is more common in the maternity ward, local health care teams should be trained in order to effectively improve the initial care of these patients. Although time of diagnosis did not affect age at surgery, it favors the planning of neonatal care and treatment of affected infants.

Time of diagnosis of oral clefts: a multicenter study.

OBJECTIVE: To determine the time of diagnosis of typical orofacial clefts in different Brazilian regions and its influence on age at surgical correction. METHOD: This was a prospective, descriptive, cross-sectional study conducted in medical centers in the Southeast, South, and Northeast of Brazil. Trained speech therapists and geneticists interviewed the parents of affected children using a previously validated questionnaire. Epi-Info and SPSS were used for data analysis. Significance level was set at 5% (p ≤ 0.05). RESULTS: The sample consisted of 215 interviews conducted in the following regions: 21.9% (47) in the Southeast, 51.1% (110) in the South, and 27% (58) in the Northeast. Monthly family income was higher in the Southeast (p ≤ 0.05). Cleft lip and palate were found in 61.4% (132) of cases, cleft palate in 20.9% (45), and cleft lip in 17.7% (38). Diagnosis occurred in the maternity ward in 75.3% (162) of cases, during the prenatal period in 14% (30), and after hospital discharge in 10.2% (22). The Southeast had a higher frequency of prenatal diagnosis (27.7%), possibly related to greater purchasing power in this region and greater availability of prenatal investigation. Of all cases diagnosed in the maternity ward, 74.4% occurred in the Northeast. However, no significant difference was found when comparing time of diagnosis, region, and age at first surgery. CONCLUSION: Considering that diagnosis is more common in the maternity ward, local health care teams should be trained in order to effectively improve the initial care of these patients. Although time of diagnosis did not affect age at surgery, it favors the planning of neonatal care and treatment of affected infants.

A clinical study of 77 patients with mucopolysaccharidosis type II.

AIM: This study aims to assess the clinical features of 77 South American patients (73 Brazilian) with mucopolysaccharidosis type II (MPS II). METHODS: Details of the patients and their disease manifestations were obtained from a review of medical records, interviews with the patients and/or their families, and physical examination of the patients. RESULTS: Mean birth weight was 3360 g, median age at onset of symptoms was 18 months and median age at diagnosis was 6 years. For the whole sample (median age, 8.2 years; range, 2.8-53.0 years), neurological degeneration, typical pebbly skin lesions, seizures and extensive dermal melanocytosis were found in 23.3, 13.0, 13.0 and 1.3% of the cases, respectively. The most frequently reported echocardiogram abnormality was mitral valve regurgitation. Refraction errors were the most common ophthalmological manifestation. The following characteristics were found to be associated with the severe form of MPS II: earlier age at biochemical diagnosis, higher levels of urinary glycosaminoglycans, language development delay, behavioural disturbances, poor school performance and mental retardation. CONCLUSION: Our results suggest that there is a considerable delay between the onset of signs and symptoms and the diagnosis of MPS II in Brazil (and probably in South America as well), and that many complications of this disease are underdiagnosed and undertreated. Therefore, the implementation of programmes aiming to increase the awareness of the disease, the availability of biochemical diagnostic tests and the provision of better support to affected patients is urgently needed.

Atypical chromosome abnormalities in acute myeloid leukemia type M4

This study reports an adult AML-M4 patient with atypical chromosomal aberrations present in all dividing bone marrow cell at diagnosis: t(1;8)(p32.1;q24.2), der(9)t(9;10)(q22;?), and ins(19;9)(p13.3;q22q34) that may have originated transcripts with leukemogenic potential.

Frequency of the BCR/ABL rearrangements and associated alterations detected by FISH during monitoring of patients taking imatinib mesylate in isolation / Freqüência do rearranjo BCR/ABL e alterações associadas detectadas por FISH no monitoramento de pacientes em uso exclusivo do mesilato de imatinibe

The introduction of imatinib mesylate as treatment of chronic myelogenous leukemia has saved many patients, but the success of therapy is hampered by resistance and possible non-destruction of the malignant clone. This article describes the cytogenetic responses and abnormal cytogenetic patterns involving the ABL and BCR genes detected by FISH in patients who use exclusively imatinib. The results showed that other alterations involving the BCR and ABL genes do not seem to be related to resistance to the drug as they occur in low frequencies and can not be associated to the cytogenetic response or to the time of treatment. Moreover, the response to imatinib seems to be individual and unpredictable, independent of the time of treatment and of its initiation after diagnosis.

A introdução do mesilato de imatinibe como tratamento da leucemia mielóide crônica tem salvado muitos pacientes, mas o sucesso da terapia tem sido prejudicado pela resistência e possível não destruição do clone maligno. Este artigo descreve a resposta citogenética e padrões citogenéticos anormais envolvendo os genes ABL e BCR detectados por FISH em pacientes em uso exclusivo de imatinibe. Os resultados mostraram que outras alterações envolvendo os genes BCR e ABL não parecem estar relacionadas à resistência à droga, elas ocorrem em baixas freqüências e podem não estar associadas à resposta citogenética ou ao tempo de tratamento. Contudo, a resposta ao imatinibe parece ser individual e imprevisível, independente do tempo e do início do tratamento após o diagnóstico.

Análise do rearranjo BCR/ABL por bandamento GTG e FISH: comparação das freqüências ao diagnóstico da LMC / Analysis of the BCR/ABL rearrangement by GTG banding and FISH: A comparison of frequencies at diagnosis of CML

A Leucemia Mielóide Crônica (LMC) é uma doença mieloproliferativa clonal caracterizada pela presença do cromossomo Philadelphia (Ph). Este cromossomo é resultante de uma translocação t(9;22) (q34;q11) que justapõe os genes BCR e ABL. A detecção do cromossomo Ph e/ou do rearranjo BCR/ABL, uma vez que este último é submicroscópico em alguns casos, é fundamental, pois não somente contribui para transformação leucêmica, mas também interfere no sucesso do tratamento. Sua detecção é, portanto, fundamental para o diagnóstico, prognóstico e terapêutica. Este trabalho teve como objetivos estudar a freqüência do cromossomo Ph ou rearranjo BCR/ABL nas células da medula óssea de pacientes portadores de LMC ao diagnóstico, com uso das técnicas de bandamento GTG e FISH, e comparar os resultados obtidos com as duas técnicas. O cromossomo Ph e o rearranjo foram observados em 100 dos casos analisados nas diferentes técnicas.Em alguns casos a freqüência do cromossomo Ph, detectado por GTG foi maior do que a do rearranjo molecular BCR/ABL detectada por FISH. A técnica de FISH também identificou alterações inespecíficas envolvendo os genes BCR e/ou ABL. Ambas as técnicas foram fundamentais para os resultados obtidos e, portanto, devem ser usadas como complementares na análise de células da medula óssea de pacientes com LMC ao diagnóstico

Sirenomelia: A case report

Caudal regression syndrome is a congenital malformation described by various degrees of developmental failure, which the most extreme and rare form is known as sirenomelia or mermaid syndrome. The associated malformations comprise anorectal, vertebral, urological, genital, and lower limb anomalies. We reported pathological findings of sirenomelia in a female stillborn with breech presentation that was born by normal vaginal delivery at 35 weeks of pregnancy following an uneventful pregnancy of a 31-year-old woman. Physical examination at birth showed normal facies, fusion of the lower limbs with bilateral presence of hip, knee, and ankle joints, sacral meningocele, omphalocele, agenesia of female external genitalia, of anus, and of cervical vertebrae. The sirenomelia etiology is still unknown but there are suggestions of genetic and teratogenic factors involvement that were not identified in the present case. The association with the agenesis of cervical vertebrae is rare and only one case described previously in the literature was reported

Leucemia mielóide crônica e o sistema Fas-FasL / Chronic myeloid leukemia and the Fas-FasL system

Com os avancos terapêuticos que incluem o mesilato de imatinibe, o transplante de medula óssea e a infusão de linfócito do doador, a perspectiva de vida dos portadores de leucemia mielóide crônica (LMC) tem aumentado significativamente e a doenca pode não ser fatal. No entanto, os mecanismos biológicos que privilegiam a selecão das células hematopoéticas malignas sobre as células normais na LMC, responsáveis pelo insucesso terapêutico em muitos casos, ainda não estão totalmente esclarecidos. Alteracões no processo de apoptose celular e escape das células leucêmicas à resposta imune antitumoral poderiam explicar, em parte, a vantagem seletiva dessas células. O processo de apoptose celular pode ser desencadeado pelas vias intrínseca ou extrínseca. A extrínseca é dependente da interacão de receptores de morte celular, como a ligacão do receptor Fas com seu receptor, o Fas ligante (FasL). A expressão diminuída de Fas e aumentada de FasL na célula leucêmica podem aumentar sua sobrevida tornando-a resistente à apoptose. Esse artigo descreve a relacão entre LMC e o sistema Fas/FasL, sua possível importância no prognóstico e escape das células leucêmicas à resposta imune.

Diagnóstico de hemoglobinopatias em recém-nascidos do Hospital de Base de São José do Rio Preto-SP / Diagnosis of hemoglobinopathies in newborn babies in Hospital de Base, São José do Rio Preto, SP, Brazil

The neonatal period is considered the most effective for the screening of hemoglobinopathies. This allows prophylaxis and prevention, improving the patient's survival and guidance of parents and heterozygote carriers. The present work aims at the early detection of abnormal hemoglobins, the establishment of standard analysis and to examine the viability of the prevention program. Blood samples were collected by heel stick and from blood cord of children born in the Hospital de Base São José do Rio Preto, from April 1998 to November 1999. Electrophoresis and cytological, biochemical, cromatographic analyses were made for abnormal hemoglobin characterization. A total of 1,478 neonatal blood samples were analyzed in which 14.62% presented with hemoglobins alterations: 3.32% had Hb S; 0.61% had Hb C; 7.44% were suggestive of alpha thalassemia; 1.55% were suggestive of beta thalassemia, and 1.70% had alpha/beta thalassemia interactions. The samples collected from the blood cord showed better results in all analyses while the blood samples collected by heel stick on filter paper, were applicable to only specific methodologies. The routine laboratory methods allowed identification of the thalassemic and variant forms, and isoelectric focusing presented sensitivity only for variant identification in this age range. The suspected cases were reassessed after six months, which permitted genetic counseling of their family members and clinic attendance. A multidisciplinary approach in programs of this kind is fundamental for its success.