TYMS gene 5'-and 3'-untranslated region polymorphisms and risk of non-syndromic cleft lip and palate in an Indian population

Dear Editor： Increased homocysteine levels due to vitamin B6 or B12 deficiency or genetic defects in folate pathway genes are associated with an increased incidence of non-syndromic cleft lip with or without cleft palate （NSCLP）tlj. Thymidylate synthase （TS） is a folate-dependent enzyme that catalyzes methylation of 2＇-deoxyuridine-5＇-monophosphate （dUMP） to 2＇-deox- ythymidine-5＇-monophosphate （dTMP）, a rate-limiting step in DNA synthesis,

Malonylcarnitine in Newborns with Non-syndromic Cleft Lip with or without Cleft Palate

Aim Malonyl-CoA is regarded as a key signaling molecule in mammalian ceils. It is converted to acetyl-CoA, and to a lesser extent, to malonyl acid and malonylcamitine （C3DC）. Availability of carnitine has been reported to be essential for the developing fetus. The objectives of the present study were to analyze associations of malonylcarnitine, acetylcarnitine （C2）, and free carnitine （CO） in subjects with orofacial clefts. Methodology We performed a retrospective analysis of carnitine concentration obtained from a newborn screening program carried out in our institution. Concentrations of whole blood malonylcarnitine, acetylcarnitine, and free carnitine were measured using tandem mass spectrometry. The study group consisted of 51 children with nonsyndromic cleft lip with or without cleft palate. In total, 106 healthy children without congenital anomalies served as controls. Cut-off points were established using likeli-hood ratio values. Results The mean concentration of malonylcarnitine in the cleft group was lower than that of the control group, 0.048 μmol.L^1 vs. 0.058 μmol.L^-1, respectively （P=-0.009）. In patients with orofacial cleft, low malonylcarnitine levels （〈0.047 μmol.L-1） were 1.7 times more predominant than in healthy individuals （P=-0.03）. The mean concentration of acetylcarnitine was also lower in affected newborns in comparison to controls, 33.8 μol.L^-1 vs. 37.8 μmol·L^-1, respectively （P=-0.026）. After analysis of acetylearnitine and free carnitine concentrations, the likelihood ratio test did not indicate valuable cut-offpoints. Conclusion The study provides initial data indicating a potential association between decreased malonylcarnitine and abnormal palatogenesis.

Updating Genetics Polymorphisms of Non-Syndromic Clefts Lip-Palates

Introduction: Non-Syndromic Clefts Lip-Palates (NSCLP/CP) are most common congenital malformation in the world, with very important psychic and social impact. Formation of NSCLP/CP arises from the interaction of environmental and genetic factors. This paper provides a review of recent progress in defining the genetic causes of NSCLP. Methods: A literature review was conducted on the Medline data by searching for the following keywords: genes, non-syndromic cleft lip-palate, and genetics of clefts lip-palates, until January 2018. Results: Various genes are identified in different population and country, with the study using case parent’s trio. The aim of this study contributes to review relative gene which has been identify in non-syndromic cleft lip and palate, and to help to have a better understanding of the inheritance pattern of this pathology and the prevention of genetic disease. Conclusion: Although three major genes have been confirmed, the genetic research is necessary to provide an understanding of the pathophysiology of the clefts lip-palates.

Assessment of Two Years of Free Surgical Treatment of Cleft Lip, Palate and Alveolar (CLPA) in the Maxillofacial Surgery and Stomatology Department of the Hospital University of Treichville-Abidjan

Introduction: Cleft lip, palate and alveolar (CLPA) are congenital malformations of the face due to a defect in the fusion of embryonic buds during the first weeks of embryogenesis. These malformations affect the upper lip, the alveolar bone and the palate. The incidence in Africa ranges from 1/2000 to 1/500 births. Their multidisciplinary management is long and costly. Thus, the help provided by humanitarian organisations during free care campaigns is welcome. Materials and Methods: This is a retrospective descriptive study conducted from August 2014 to July 2016 in the Maxillofacial Surgery and Stomatology Department of the Treichville University Hospital in Abidjan, Côte d’Ivoire. The objective was to describe the epidemiological, clinical and therapeutic aspects of CLPA during a humanitarian campaign for free care. Results: 51 cases of CLPA were operated on. Males were involved in 54.9% of the cases, i.e. a sex ratio of 1.2. The average age of the patients at the time of the operation was 3.44 years with extremes of 3 months and 52 years. Patients with low socioeconomic status represented 84.3% of the cases. Cleft lips (31.4%) and cleft palates (33.33%) predominated. For cleft lips, unilateral forms were the most frequent (73.5%) and the left side was most often affected (59.2%). The most common surgical techniques used were MILLARD cheiloplasty for cleft lips (79.36%) and Dorrance pushback for cleft palates (78.05%). The postoperative course was simple in the majority of cases (80.47%). Patients and/or parents were satisfied with the postoperative results in over 90% of cases. Discussion: Cleft lip and palate are common. Their management by humanitarian missions through mass campaigns allows us to receive a large number of patients affected by this pathology who are treated with a high satisfaction rate. Conclusion: The characteristics of cleft lip and palate in this study are in many respects identical to those described in the literature, but with some differences specific to Africa, notably the absence of antenatal diagnosis and the advanced age at the time of treatment.

Rhinoplasty in Cleft Lip and Palate

Various surgical techniques and approaches have been described to repair cleft nose deformities. It is necessary to consider that since it is a congenital deformity, surgical management must consider the patient’s growth process, as well as the healing itself from the surgery. The present study aims to evaluate an alternative solution to secondary rhinology, aesthetic, and functional alterations to unilateral or bilateral cleft lift palate to minimize deformities and ensure good results. 11 patients were studied between 1995 to 2002, ten male and one female. In 8 cases, the patients had a history of cleft lip and palate on the left side, 2 patients with cleft lift palate on the right, and 1 patient with bilateral cleft lip and palate. 100% of the patients increased their naso-labial angle. This improved their appearance and structure, starting from a preoperative arithmetic mean of 39 degrees to a postoperative arithmetic mean of 96 degrees. Such intervention increased the naso-labial angle by 57 degrees. At the base of the nose, it was possible to improve the inclination of the alar line (line B) in 10 of the cases with a variation of 2 to 3 mm in relation to the perpendicular line A and only one case remained with the same inclination.

The Craniofacial Morphology in Adult Patients with Unoperated Isolated Cleft Palate

Objective： To address the effect of intrinsic factors on craniofacial growth by analyzing the craniofacial morphology of unoperated isolated cleft palate in Chinese adult. Materials and Methods： This study included 37 nonsyndromic isolated cleft palate and 39 age and gender matched non-clefts. Twenty-six cephalometric measurements were employed to evaluate the facial morphology. Independent samples T test and Mann- Whitney U were used for comparison. Significant difference was defined at 95% level. Results： Data from this study showed patients with unoperated isolated cleft palate have a reduced maxillary sagittal length （ANS-PMP, A-PMP, P〈0.05）, a smaller ANB angle （ANB, P〈0.05） and a retrusive ANS point （S-N-ANS, P〈0.05; Ba-N-ANS, P〈0.05）. Measurements descripted position of maxilla （S-Ptm, P〉0.05）, depth of bony pharynx （Ba-PMP, P〉0.05）, anterior and posterior maxillary height （N-ANS, P〉0.05; R-PMP, P〉0.05） and mandible morphology （including linear measurements and angle measurements） did not show any significant difference between case and control groups. Conclusions： Patients with isolated cleft palate were characterized by maxillary retrusion. Mandible morphology and cranial basal morphology in isolated cleft palate showed no significant difference with nonclefts. Patients with isolated cleft palate are more vulnerable to cross bite than nonclefts. Intrinsic deficiencies did detrimental effect on maxilla sagittal length, but did no detrimental effect on maxilla position, mandible size and position.

Accurate diagnosis of prenatal cleft lip/palate by understanding the embryology

Cleft lip with or without cleft palate(CP) is one of the most common congenital malformations. Ultrasonographers involved in the routine 20-wk ultrasound screening could encounter these malformations. The face and palate develop in a very characteristic way. For ultrasonographers involved in screening these patients it is crucial to have a thorough understanding of the embryology of the face. This could help them to make a more accurate diagnosis and save time during the ultrasound. Subsequently, the current postnatal classification will be discussed to facilitate the communication with the CP teams.

BHMT Gene Polymorphisms as Risk Factors for Cleft Lip and Cleft Palate in a Chinese Population

Objective Convincing evidence suggests a link between increased risk of nonsyndromic cleft lip with or without cleft palate （NSCL/P） and low intake of folic acid by the mother during pregnancy. The present study was designed to explore if genetic variation in the betaine‐homocysteine methyltransferase （BHMT） gene contributes to NSCL/P. Methods DNA was obtained from 166 individuals with NSCL/P and 285 healthy subjects. Three known single nucleotide polymorphisms （SNPs） present in the BHMT gene （rs651852, rs3797546, and rs3733890） were investigated by real‐time PCR‐based TaqMan genotyping. Results Neither allelic nor genotypic association was found between NSCL/P and SNPs rs651852 and rs3733890. SNP rs3797546 did not show allelic association with NSCL/P; however, a higher proportion of NSCL/P patients carry the CC genotype compared with the TT＋CT genotype （P=0.020, OR=2.10, 95% CI=1.11‐3.95）. Conclusion Our study suggests that polymorphism rs3797546 in the BHMT gene may confer genetic risk of NSCL/P in a recessive manner.

RETINOIC ACID DOWN-REGULATES BONE MORPHOGENETIC PROTEIN 7 EXPRESSION IN RAT WITH CLEFT PALATE

Objective To evaluate the effects of retinoic acid （RA） on expression of bone morphogenetic protein 7 （ BMP-7 ） in rat fetus with cleft palate, and the effects of RA on proliferation and apoptosis of osteoblasts. Methods All-trans RA （ATRA） was used to induce congenital cleft palate in Wistar rat. BMP-7 mRNA expression in maxillary bone tissue of fetal rats was measured by Northern blotting analysis. Flow cytometry and MTF assay were used to measure the apoptosis and proliferation of ATRA-treated MC-3T3-E1 cells. BMP-7 mRNA and protein expressions in ATRA-treated MC-3T3-E1 cells were detected by RT-PCR and Western blotting analysis. Remilts ATRA could induce cleft palate of rat fetus. The incidence rate of cleft palate induced by 100 mg/kg AT-RA （45.5%） was significantly higher than 50 mg/kg ATRA （ 12.5%, P 〈 0. 05 ）. BMP-7 mRNA expression decreased in maxillary bone tissue of rat fetus with cleft palate. MC-3T3-E1 cells proliferation treated with 1 × 10^-6 mol/L ATRA decreased by 60%, the cell apoptosis increased by 2 times. BMP-7 mRNA and protein levels in MC-3T3-E1 cells treated with 1 × 10^-6 mol/L ATRA decreased by 60% and 80%, respectively, compared with ATRA-untreated cells （ P 〈 0.05 ）. Conclusions BMP-7 may play an important role in embryonic palate development. RA may possess the ability to down-regulate cell proliferation through regulation of BMP-7 gene expression.

Genome-wide and Interaction Linkage Scan for Nonsyndromic Cleft Lip with or without Cleft Palate in Two Multiplex Families in Shenyang,China

Objectives To identify the loci involved in nonsyndromic cleft lip with or without cleft palate （NSCL/P） in Northern Chinese people in Shenyang by using genomewide and interaction linkage scan.Methods Two multiplex families in Shenyang from North China were ascertained through probands with NSCL/P.Blood of every member was drawn for DNA extraction and analysis.Genotypes were available for 382 autosomal short tandem repeat （STR） markers from the ABI Prism Linkage Mapping Set version 2.5.Linkage between markers and NSCL/P was assessed by 2-point parametric LOD scores,multipoint heterogeneity parametric LOD scores （HLODs）,and multipoint nonparametric linkage score （NPL）.Results The initial scan suggested linkage on Chromosomes 1,2,and 15.In subsequent fine mapping,1q32-q42 showed a maximum multipoint LOD score of 1.9（empirical P=0.013） and an NPL score of 2.35 （empirical P=0.053）.For 2p24-p25,the multipoint NPL increased to 2.94 （empirical P=0.007）.2-locus interaction analysis obtained a maximum NPL score of 3.73 （P=0.00078） and a maximum LOD score of 3 for Chromosome 1 （at 221 cM） and Chromosome 2 （at 29 cM）.Conclusion Both parametric and nonparametric linkage scores greatly increased over the initial linkage scores on 1q32-q42,suggesting a susceptibility locus in this region.Nonparametric linkage gave a strong evidence for a candidate region on chromosome 2p24-p25.The superiority of 2-locus linkage scores compared to single-locus scores gave additional evidence for linkage on 1q32-q42 and 2p24-p25,and suggested that certain genes in the two regions may contribute to NCSL/P risks with interaction.

LncR NA H19 and Target Gene-mediated Cleft Palate Induced by TCDD

This study investigated the role of long non‐coding RNAs（lnc RNAs） in the development of the palatal tissues. Cleft palates in mice were induced by 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin（TCDD）. Expression levels of long non‐coding RNA H19（lncR NA H19） and insulin‐like growth factor 2（IGF2） gene were measured by quantitative real‐time polymerase chain reaction（q RT‐PCR）. The rate of occurrence of cleft palate was found to be 100% by TCDD exposure, and TCDD could cause short upper limb, cerebral fissure, webbed neck, and short neck. The expression levels of lnc RNA H19 and IGF2 gene specifically showed embryo age‐related differences on E13, E14, and E15 in the palatal tissues. The expression levels of lnc RNA H19 and IGF2 gene showed an inverse relationship on E13, E14, and E15. These findings demonstrated that lnc RNA H19 and IGF2 can mediate the development of mouse cleft palate.

Molecular basis of cleft palates in mice

Cleft palate, including complete or incomplete cleft palates, soft palate clefts, and submucosal cleft palates, is the most frequent congenital craniofacial anomaly in humans. Multifactorial conditions, including genetic and environmental factors, induce the formation of cleft palates. The process of palatogenesis is temporospatially regulated by transcription factors, growth factors, extracellular matrix proteins, and membranous molecules; a single ablation of these molecules can result in a cleft palate in vivo. Studies on knockout mice were reviewed in order to identify genetic errors that lead to cleft palates. In this review, we systematically describe these mutant mice and discuss the molecular mechanisms of palatogenesis.

AN ANALYSIS OF ACOUSTIC CHARACTERISTICS OFCLEFT PALATE SPEECH WITH COMPUTERIZED SPEECH SIGNAL PROCESSING SYSTEM

The acoustic characteristics or the chinese vowels of 24 children with cleft palate and 10 normal control children were analyzed by computerized speech signal processing system (CSSPS),and the speech articulation was judged with Glossary of clert palate speech(GCPS).The listening judgement showed that the speech articulation was significantly different between the two groups(P<0.01).The objective quantitative measurement suggested that the formant pattern(FP)of vowels in children with cleft palate was different from that of normal control children except vowel[a](P< 0.05).The acoustic vowelgraph or the Chinese vowels which demonstrated directly the relationship of vocal space and speech perception was stated with the first formant frequence(F1)and the second formant frequence(F2).The authors conclude that the values or F1 and F2 point out the upward and backward tongue movement to close the clert, which reflects the vocal characteristics of trausmission of clert palate speech.

Perioperative Management of Cleft Palate Repair in a Patient with Williams Syndrome: A Case Report

The Background: Williams syndrome is a neurodevelopmental multisystem genetic disorder characterized by dysmorphic features and a wide range of congenital cardiac, renal, musculoskeletal anomalies. Presence of cleft palate in these patients is an unusual condition. More than 22 cases of sudden cardiac death reported in literature especially during anesthetic management in the patients of Williams syndrome. So these patients present major perioperative challenges to anesthesiologist. The authors report successful anesthetic care for repair of cleft palate in presence of uncorrected patent ductus arteriosus (PDA) and mild supraventricular aortic stenosis (SVAS) in a 3.5-year-old female child diagnosed with Williams syndrome. Case report: Diagnosed case of Williams syndrome with characteristic facial dysmorphic features, SVAS and rare malformation of PDA, was scheduled for cleft palate repair. After keen preoperative assessment, general anesthesia was administered for the procedure with continuous monitoring for vital parameters. Because of dysmorphic features, difficulty was faced during intubation. Otherwise intraoperative procedure was uneventful. Postoperative analgesia was managed with intravenous paracetamol. Conclusion: Being a multisystem disorder, each patient of WS requires meticulous preoperative evaluation and high level of intraoperative and postoperative continuous monitoring regardless of any surgical procedure. Surgical correction of cleft palate in such patients demands very high anesthetic and surgical skills as both share common airway. Present case report highlights the significance of aggressive perioperative management in WS which can result in successful outcome.

A Case-control Study of Environmental Risk Factors for Nonsyndromic Cleft of the Lip and/or Palate in Xuzhou,China

In this study, we sought to determine the association between environmental factors and nonsyndromic cleft of the lip and/or palate （NSCLP） to understand the etiology of the disease. A total of 200 NSCLP cases and 327 controls were recruited at the Maternal and Child Health Hospital of Xuzhou City. We conducted face-to-face interviews with the mothers of both cases and controls. The factors increasing the risk of NSCLP were a positive family history [odds ratio （OR）=56.74], pesticide exposure （OR=8.90）, and indoor decoration pollution （OR= 4.32）. On the other hand, the factors decreasing the risk of NSCLP were a high education level （OR=0.22） and supplementation of folic acid （OR=0.23） and multivitamins （OR=0.16）. Positive family history, pesticide exposure, and indoor decoration pollution are associated with the risk of NSCLP. In contrast, high education level and folic acid and multivitamin supplementation are protective factors against NSCLP.

Comparison of Congenital Tooth Deficiencies Seen in Permanent Teeth in Individuals with Unilateral Cleft Lip and Palate to Those without Cleft Lip and Palate

Objective: To compare the congenital tooth deficiencies seen in permanent dentition in individuals with unilateral cleft lip and palate (UCLP) to non-cleft individuals with Angle Class I malocclusion. Method: The study was performed on orthopantomograph films of 50 individuals with UCLP aged between 12 - 16 years and 50 individuals with Angle Class I malocclusion individuals aged between 13 - 15 years. Individuals with UCLP;32 clefts were on the left side and 18 clefts were on the right side. Permanent third molar teeth deficiency was excluded from the study. Results: In 50 individuals with UCLP;35 (70%) upper lateral incisors were congenitally deficient in the cleft area, while 15 (30%) missing lateral teeth were found in the non-cleft side. In control group, 12 (24%) of 50 patients had congenital lateral incisor deficiency. Congenital deficiency of upper lateral incisor in UCLP;the cleft area was higher than the non-cleft side (p < 0.01). The difference was statistically important when compared with the control group (p < 0.001). In 50 individuals with UCLP;while 27 (54%) of second premolar teeth were congenitally deficient in cleft side, 23 (46%) were missing in non-cleft side. In control group, 18 (36%) congenital second premolar deficiency was detected. However, second premolar congenital deficiency was higher in UCLP group when compared to control group (p < 0.01). Conclusion: The deficiency of the lateral incisors in the cleft side is more often congenitally deficient than upper second premolar teeth and this should be considered in the treatment planning from an early age.

Effect of Cleft Lip and Palate Repair on Craniofacial Morphology in Patients with Unilateral Complete Cleft Lip and Palate.

目的 :通过对替牙期正常儿童及单侧完全唇腭裂 (UCLP)患者X线头影测量侧位片进行比较研究 ,了解唇腭裂及其手术对颌骨发育的影响 ,探索继发畸形的发病机制。方法 :将替牙期的正常儿童及UCLP患者分为四组 :①正常对照组 (Normal) ,本组包括 48名替牙期正常湖北地区儿童 ;②未手术组 (NO) ,本组 10名UCLP患者未经任何手术治疗 ;③唇裂修复组 (OL) ,34名UCLP患者已行唇裂修复 ,腭裂暂未手术 ;④唇腭裂修复组 (OLP) ,16名UCLP患者已行唇、腭裂修复。各受试对象拍摄X线头影测量侧位片 ,测量描述颌骨的形态特征及生长方向 ,对各组之间测量结果进行统计学分析。结果 :未行手术治疗的U CLP患者颌骨形态除腭部先天性发育不足外 ,颌骨侧位头影测量与正常无显著性差异 ;OL和OLP组较正常存在严重的颌骨畸形 ,主要表现为上颌后缩、位置后移 ,同时下颌骨向后下旋转移位 ;OL和OLP组上颌后缩的程度无显著性差异。结论 :UCLP患者颌骨具有趋于发育正常的生长能力 ;手术是导致颌骨继发畸形的主要因素 ;对于单侧完全唇腭裂患者 。

Value of acoustic perceptual method for analysis of compensatory articulation errors in postoperative patients with cleft palate

Objective To establish an acoustic perceptual method analyzing the compensatory articulation errors in children with operated cleft palate via the formants of Chinese pure vowels. Methods The first three formants which represent vocal transmission character in Chinese pure vowels of 84 subjects with operated cleft palate, were measured by Computerized Speech Signal Processing System (CSSPS). The Chinese vowel graph of postoperative patients with cleft palate was stated by the first formant frequencies (F1) and the second formant frequencies (F2) of the Chinese pure vowels between the two groups. Results Values of F1 and F2 of vowels except [a] in the poor articulation group (Group A) were significantly different from those in the good articulation group (Group B) (P<0.05 or P<0.01). As compared with that in Group B, the vowel graph demonstrated the decreased perceptual distances in Group A. These findings indicated that there might still be the backward movements of tongue, perverted mandibular movements and disharmonious lip movements in addition to the velopharyngeal insufficiency (VPI) in Group A. Conclusion The speech articulation of children with repaired cleft palate should be gained by correcting the aberrant compensatory articulation errors in the condition of velopharyngeal competence. Computerized Speech Signal Processing System (CSSPS), which is regarded as the content of objective quantitative measurement, is a precise, simple, reliable and atroumatic technique for children with cleft palate to analyze pathological compensatory articulation errors.

Evaluation of Speech Intelligibility of a Cleft Palate Patient with Speech Prosthesis before and after Palatographical Analysis and Prosthesis Modification

The aim of the study was to evaluate the alterations in speech intelligibility in a cleft palate patient, before and after extending and modifying the palatal contour of the existing prosthesis using a correctable wax recording. An eight-year-old girl studying in second grade with a velopharyngeal defect using an obturator reported to the outpatient clinic complaining of lack in clarity of speech. The existing prosthesis was lacking a speech bulb hence it was decided to add the speech bulb to the existing prosthesis and evaluate the speech. Even after the use of speech bulb it was observed that she was unable to pronounce the vowels and words like shoe, vision, cheer, etc. clearly. Hence, a palatography was done using a correctable wax technique and the existing prosthesis was altered accordingly. Great improvement in speech, mastication, and velopharyngeal function was achieved after the palatography alteration of the existing prosthesis.

Evaluation of Maxillary Dental Arch Widths in Individuals Having Unilateral and Bilateral Cleft Lip and Palate Using Three-Dimensional Digital Models

Aim: Maxillary dental arch widths were evaluated in individuals having unilateral (UCLP) and bilateral (BCLP) cleft lip and palate (CLP) using three-dimensional (3D) digital models. Material and Method: The study had been conducted on 80 individuals aged between 14 - 17 years having UCLP and BCLP. 40 of the individuals had UCLP, whereas 40 had BCLP. The maxillary dental models taken from patients before the treatment were scanned using Orthomodel Programme (v.1.01, Orthomodel Inc., Istanbul, Turkey) to obtain 3D imagery. Student’s t-test was used in order to assess the data obtained by using SPSS software version 22.0. Results: In BCLP, the average inter-canine distance was 17.44 ± 1.31 mm, the average inter-molar distance was 36.57 ± 1.12 mm, while inter-canine/inter-molar ratio was 0.47. Whereas in UCLP, it was 25.10 ± 0.63 mm, 42.20 ± 0.53 mm and 0.59. The inter-canine distance in UCLP was found to be large enough to be statistically significant (p 0.05), even though there were differences in inter-molar widths. Conclusion: For the stable orthodontic treatment results, one of the most important points is arch form and widths to be coherent with each other. In our study, the increase of inter-canine distance seen in UCLP indicates that in the cleft region, the maxillary arch is inclined over to the back, while the same situation in BCLP suggests that the maxillary segments are collapsed inside. The difference in the arch is highly affected by the primary surgical treatment.