Cleft lip and palate in mice treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin: a morphological in vivo study

It is well-known that TCDD (2,3,7,8, tetrachloridedibenzo-p-dioxin) induces cleft palates (CPs) in pregnant C57BL mice. However, it is unclear if TCDD is a possible teratogen for cleft lip. We examined maxillofacial malformations including cleft lip in three animal strains: A/J mice, C57BL/6J mice and ICR mice. The A/J mouse develops cleft lip and palate spontaneously at a 5-10% rate. TCDD was administered in olive oil on gestation day (GD) 12.5 with gastric tubes at 10 microg/kg, 20 microg/kg, or 40 microg/kg to examine the dose-response, and on a single day from GD 8.5-14.5 to examine the timing effects of TCDD administration on lip and palate formation. Furthermore, the palatal shelf movements during GD 8.5-14.5 were observed with a stereoscopic microscope. All embryos had cleft palates when the TCDD was administered just before palatogenesis (GD11.5-GD12.5). With respect to the TCDD effects, there were large differences among the strains. In the A/J mice, the difference between a lethal dose and a dose that could induce a cleft palate was close. Cleft lips were not induced, even when the TCDD was given just before labiogenesis. Morphologically, both palatal shelves contacted perfectly along their lengths, but separated and formed cleft palates. In conclusion, TCDD is a strong inducer of cleft palates, and interferes with the fusion phase of the secondary palate, but has no effect on the lip.     

Spontaneous Intrauterine Repairment of Cleft Palate Induced by Amniocentesis in Rats

On day 15 of gestation, 180 fetal sacs in 43 pregnant Charles Foster rats were subjected to amniocentesis producing a series of total or partial clefts in the hard and soft palate regions. Fetuses recovered on days 19, 20 or 21 showed a gradual decline in the incidence of total clefts with increasing gestational age, i.e. 34% on day 19, 29% on day 20 and 13% on day 21, suggesting spontaneous ‘repairment’ of the clefts induced by amniotic sac puncture. Prenatal ‘repairment’ at times leaving residual partial clefts in the hard palate, suggested delayed rotation of the palatine shelves following reaccumulation of lost quantity of amniotic fluid and withdrawal of tongue obstruction. The experiment suggested that the palatine shelves retain intrinsic ‘shelf force’ for a much longer period than thought earlier. However, a gradual increase in the incidence of residual partial cleft with increasing gestational age, i.e. 45% on day 19; 50% on day 20 and 67% on day 21, respectively, suggests only incomplete (partial) repair in most of the cases probably due to paucity of time.     

COL2A1基因变异相关腭裂1例报告并文献复习

目的探讨腭裂患儿的临床特征及致病基因。方法回顾分析1例COL2A1基因变异腭裂患儿的临床资料,并复习c.2292 delA变异相关腭裂相关文献。结果患儿男,生后即发现上腭畸形,临床表现有双眼略凸出、哭声低哑、喉中痰鸣、呼吸阵发性急促有吐沫、下颌小、舌短、软腭及悬雍垂裂、硬腭部分裂开。全外显子组基因靶向捕获-高通量测序示患儿COL2A1基因存在c.2292 delA移码缺失变异。文献复习发现,COL2A1基因移码变异已在人类基因变异数据库(HGMD)和ClinVar中报道,但本例患儿的变异位点未有报道,在正常人群数据库gnomAD、千人数据库和ExAC数据库中也均未被收录,为罕见变异。结论COL2A1基因变异c.2292 delA相关腭裂较罕见,基因检测可协助诊断。     

A Review of Unilateral Cleft Lip and Palate Surgery*

ABSTRACT Operative techniques for unilateral cleft lip and palate have undergone continuous development, with improvement and modification, during the last three decades. The author's operation began with Tennison's method and gradually changed into a narrower and long triangular flap method. The edge of the orbicularis oris muscle on the cleft side is tucked under the center of the philtral dimple to obtain muscles linkage and a philtral ridge on the affected side. The longitudinal line is sutured with meticulous subcuticular stitches using 6–0 Nylon, resulting in a neat scar. Relationship between vertical height of lip and width of triangular flap was examined in follow-up studies. Flap width increased rapidly for one year after surgery and then slowed down to a pace proportionate with lip height. Although ratio of flap width to height increased from 27% to 40%, the balance of both sides of the lip did not, as had been thought might be possible, change. Correction of the cleft lip nose by the triangular flap method is rather difficult. It is considered natural for many surgeons to prefer a combination of the rotation-advancement flap and small triangular flap. Much remains to be discussed regarding cleft palate treatment. Author prefers closure of the hard palate using a vomer flap in the primary cleft lip operation because the most effective push-back of the palate, without fistulae, is achieved. Underdevelopment of lip and maxilla should be evaluated from many aspects, and with a long-term view.     

Clinical Analysis of the Cleft Patterns of Lip and Palate

The patterns in the cleft lip and palate were classified in great detail, and the point of center of the clefts in this disease was speculated. The purpose of the present study was to establish the basic data for classifications of epidemiological surveys in the future. The subjects were 377 patients with cleft lips and palates who visited the Second Department of Oro-Maxillo-Facial Surgery, Aichi-Gakuin University Hospital. The models of cleft lip and palate divided into 17 segments prepared for an analysis of cleft patterns. And as consequence, the following results were obtained.     

Clinical Aspects of Cleft Lip and Cleft Palate Patients Treated at Kanazawa Medical University Hospital from 1974 to 1993*

ABSTRACT Currently, clefts of the lip, alveolus, and palate are thought to be the most common congenital malformations. In this study, we examined 1,565 cases of clefts we treated over the last 18 years, in order to summarize the clinical features and to evaluate the treatment outcomes. This study involved 589 cleft lip (CL) cases, 576 cleft lip and palate (CLP) cases, 286 cases of cleft palate (CP), 81 cases of submucous CP and 33 cases of CL and submucous CP. The familial incidence was 11.1% (174 cases). Overall, the average incidence of associated congenital abnormalities was 27.9%. Approximately half of the patients with CP alone or with submucous CP were found to have additional congenital malformations. Standeard surgical techniques for unilateral and bilateral CL cases were the Skoog's and Millard's methods, respecively, which resulted in satisfactory outcomes. As a primary operation for CP, pushback procedure was performed using a partial mucosal flap from the palate. Using this procedure 96.5% of 455 cases achieved satisfactory nasopharyngeal closure. The main objective in the treatment of CL patients is not only the repair of growing tissues but also appropriate morphological, functional and psychological reconstruction for the purpose of correcting growth inhibitory problems caused by anatomical malposition. Surgical procedures for CP patients are designed to meet several needs: correction of anatomical abnormalities of the palate and pharynx, achievement of proper nasopharyngeal closure necessary for normal phonation, prevention of growth disturbances of the maxilla, improvements in hearing and auditory tube function, and normalization of occlusion.     

Effects of the Ay Gene on the Sensitive Periods of Hydrocortisone-Induced Cleft Palate and Palatal Slit in Mice

The effects of the A^y gene on the sensitive periods of hydrocortisone-induced cleft palate and palatal slit were investigated in C57BL/6 (a/a) and its congenic strain of C57BL/6-A^y (A^y/a) mice by using the single-dose administration method. Matings were conducted as follows (femalex male): Group I, a/a × a/a; Group n, a/a ×A^y/a; and Group III, A^y/a × a/a. Pregnant females were subcutaneously given a single dose of 200 or 400 mg/kg of hydrocortisone on day 11, 12, 13 or 14 of pregnancy and killed on day 18 for examination of palatal defects of live fetuses. Cleft palate was produced in all groups by treatment on days 11–13 of pregnancy at a dose of 200 mg/kg without any increase in fetal mortality. In Group I, the maximum incidence was observed on day 11 and followed by a rapid decrease thereafter. In Group U, in which half of the fetuses were expected to carry the A^y gene, the incidence still remained high on day 12, although it was decreased to the level of Group I by day 13. In Group HI, in which maternal mice as well as half of their fetuses carried the A^y gene, significantly higher incidences were observed on days 12 and 13 with the maximum value on day 12 when compared with those in Groups I and/or II. Palatal slit was also induced in all groups on each of treatment days at a dose of 200 mg/kg. Although the incidences in both Groups I and II were high on days 11 and 12 and gradually decreased thereafter, the value on day 13 was significantly higher in Group II than in Group I. In Group IE, the maximum incidence was found on day 12 and the values on days 13 and 14 were still significantly higher than those in Groups I and/or II. These results indicate that the sensitive periods of hydrocortisone-induced cleft palate and palatal slit are prolonged into the later stage of embryonic development as a result of the A^y-gene effects.     

Prevalence of cleft lip and/or palate in children from Lodz between years 1981–2010

Congenital malformations constitute a serious problem of both medical and social nature. Cleft lip and/or palate represent the most common congenital anomaly of the face that is why it is essential to know the real frequency of the described phenomenon. The aim of this paper is to determine the frequency of cleft lip and/or palate and the types of malformations that occurred in Lodz city between the years 1981–2010. Our clinic has been carrying on the studies concerning the incidence of cleft lip and/or palate since 1981. The Polish Registry of Congenital Malformations has been operating in Poland since 1 April 1997. The team has managed to obtain data, from the Registry, concerning the total number of all live born infants and the number of children with cleft lip and/or palate, who were born in Lodz, between 1998 and 2010. In years 1981–2010, 319 children, in 210 952 live born infants, were born with cleft lip and/or palate in Lodz. The isolated cleft palate was observed more frequently in girls and the unilateral cleft of lip and palate in boys. In all three decades palate clefts are more common whereas bilateral lip, alveolus and palate clefts are more infrequent. A small tendency to decrease in actual cleft lip and/or palate frequency among children, in the period of 30 years, is observed in Lodz. Over the years it has still been observed that the isolated cleft palate is the most common type of defect.     

Ectrodactyly,ectodermal dysplasia,cleft lip and palate (EEC)—a rare syndrome

A case of ectrodactyly, ectodermal dysplasia and cleft lip and palate (EEC) syndrome featuring bilateral, cleft lip and palate, ectrodactyly, ectodermal dysplasia, occular and renal abnormalities and spina bifida of 5th lumbar vertebra is presented. The patient also had certain dermatoglyphic abnormalities, however, the chromosomal complement was normal.     

Paediatric and ophthalmologic observations in offspring of alcohol abusing mothers

The offspring of nine women who had abused alcohol and drugs during pregnancy were studied. Of the 30 children, 10 had fetal alcohol syndrome (FAS) or fetal alcohol effects (FAE). Ophthalmological impairments associated with FAS—impaired vision, optic nerve hypoplasia, cataract, increased tortuosity of retinal vessels—were common. The severity of teratogenic lesions varied among the children and was often related to the level of abuse during pregnancy. Most women were unable to take care of their children during periods of substance abuse. All children with FAS/FAE had learning difficulties. Four mothers abstained from alcohol or drugs during some of their pregnancies and gave birth to children without birth defects.     

Regional heterogeneity in the developing palate: morphological and molecular evidence for normal and abnormal palatogenesis

Development of the mammalian secondary palate involves the growth, elevation, medial elongation and midline fusion of palatal shelves. Recent morphological and molecular studies on palatogenesis suggest that the developing palate is not a homogeneous organ but each part may behave differently during organogenesis. Especially, some key molecules involved in palate development have been shown to exhibit heterogeneous patterns of expression in the palatal tissue. Therefore it seems necessary to recognize the regional heterogeneity of the developing palate along the dorsoventral and anteroposterior axes when analyzing the mechanisms of normal and abnormal morphogenesis. Based on recent studies, we discuss the issue of the regional heterogeneity in the fetal palate and propose a principle that divides the fetal palate into several regions from the morphological and molecular standpoint.     

Effect of Maternal Treatment with Methylmercury on the Manifestation of Cleft Lip in CL/Fr Mice

Abstract The interaction between methylmercury teratogenesis and manifestation of cleft lip and palate (CL(P)) of genetic origin was experimentally investigated in CL/Fr mice. Pregnant mice were given methylmercuric chloride orally on day 10 of pregnancy at doses of 5, 10, 15 and 20 mg/kg. Animals were put to death on day 18 of pregnancy and the fetuses were examined for malformations, especially facial anomalies. About 23% of fetuses manifested CL(P) in the control group. After maternal treatment with methylmercuric chloride, the incidence of fetuses with CL(P) did not remarkably change in groups of 5 mg/kg (about 30%) and 10 mg/kg (about 24%), but decreased significantly (p<0.01) in groups of 15 mg/kg (about 12%) and 20 mg/kg (about 12%). The average numbers of both early and late fetal deaths in the mercury-treated groups were not significantly different from those in the control group (p > 0.05): This suggested that the decreased incidence of fetuses with CL(P) was not attributable to the preferential mortality of embryos with CL(P). The incidence of isolated cleft palate (CP) was about 1, 5,40 and 80% after maternal treatments of 5,10,15 and 20 mg/kg methylmercuric chloride, respectively, showing the distinct threshold. No significant difference in mercury concentration was observed among normal fetuses, fetuses with CL(P) and with CP within any dose group.     

Growth in children with Pierre Robin sequence and isolated cleft palate

Postnatal height and weight growth were assessed in 50 children (20 boys) with Pierre Robin sequence and in 58 children (27 boys) with isolated cleft palate, born in 1967–86. The height and weight measurements from birth to 12 years were collected retrospectively from child health centers and schools. The current Finnish relative weight and SD scores for height were used for growth assessment. The birth size of children with Pierre Robin sequence did not differ from those with isolated cleft palate or from healthy children, on the basis of Finnish norms. During the first year after birth, children with Pierre Robin sequence were shorter and lighter than those with isolated cleft palate but later caught up with them and the Finnish norms. Children with Pierre Robin sequence born prematurely or with associated anomalies showed more deficient growth.     

Feeding behaviour of infants with cleft lip and palate

AIM: To determine whether the feeding behaviour of infants with cleft lip and palate is improved with a type-P teat, which is widely used in Japan by such infants, compared with a standard teat. The difference in intra-oral movements between the type-P teat, modified for the evaluation of feeding behaviour, and an unmodified type-P teat was also compared using ultrasonography. METHODS: In part 1 of the study, 15 infants aged 2 to 3 mo and 7 infants aged 2 wk were evaluated for sucking pressure, expression pressure, frequency and duration of sucking. All the infants had a complete unilateral cleft lip and palate without any other abnormalities. In part 2, an ultrasonographic analysis of intra-oral movement was done for 5 infants enrolled in part 1 of the study. RESULTS: Sucking pressure did not occur in all infants. It was found that feeding efficiency improved with the type-P teat compared with the standard teat. The expression pressure with the type-P teat was significantly higher than that with the standard teat, and the feeding frequency with the type-P teat was lower than that with the standard teat. CONCLUSION: A type-P teat is suitable for infants with cleft lip and palate who have sucking difficulties. However, a type-P nipple with a squeezable bottle does not fully solve the feeding problems of infants with cleft lip and palate. New artificial teats that allow a higher expression pressure are desirable, and the measurement of the expression pressure may be helpful in the evaluation of artificial nipples.     

Induction of cleft palate in aniline hydrochloride-treated rats: Possible effect of maternal methemoglobinemic hypoxia

ABSTRACT  Aniline hydrochloride (AH), a methemoglobin formation-stimulating substance, at a dosage level of 520 mg/kg which does not induce apparent fetal death, was injected subcutaneously into pregnant rats once on day 14, 15 or 16 of gestation in order to assess the stage specificity of cleft palate induction. Also, doses of 260, 390, 520 and 650 mg/kg were administered to pregnant rats on day 15 of gestation, and the dose-response relationships with respect to fetal cleft palate and maternal methemo-globinemia induction were studied. In the stage-specificity study, paleness, decreased body weight gain and elevated methemoglobin concentration were noted in the dams treated with AH. Upon fetal examinations, although reduced body weight was noted in all AH-treated groups, cleft palate was observed only in fetuses from those dams treated on day 15 of gestation. In the dose-dependency study, AH induced maternal methemoglobinemia, decreased fetal body weight and increased the incidence of cleft palate dose dependently when administered at dosage levels of 260, 390, 520 and 650 mg/kg on day 15 of gestation. Additionally, administration of methylene blue, a methemoglobinemia-preventing substance, to the AH-treated dams ameliorated maternal methemoglobinemia and reduced the incidence of fetal cleft palate. In summation, it is considered that AH stage-specifically induces cleft palate in rats and that cleft palate is caused not by a direct teratogenic effect of AH but by maternal hypoxia due to methemoglobinemia.     

Effects of N-acetyl-L-cysteine on Teratogenicity of Cadmium in Mice

ABSTRACT Embryotoxicity and teratogenicity of cadmium (Cd) and modulation of its effects by N-acetyl-L-cysteine (NAC) were evaluated in mice. Pregnant ICR mice were intraperitoneally injected with 3.5 mg/kg of CdCl₂ on day 10 or 11 of gestation (vaginal plug = day 0). Pregnant mice were pretreated with 160 mg/kg of NAC intravenously 2 hours before dosing with CdCl₂. Pregnant mice were killed on day 17 of gestation. Fetuses were examined for external malformations, especially limb malformations, cleft palate and abnormal palatal rugae. There was little difference in body weight gain of dams during the gestation period in the groups treated with NAC plus Cd as compared with the groups treated with Cd alone. Pretreatment with 160 mg/kg of NAC decreased the fetal mortality, incidence of cleft palate and abnormal palatal rugae induced by Cd on day 11. On day 10, pretreatment with NAC decreased the incidence of Cd induced abnormal palatal rugae. These results clearly indicate that NAC exerts protective effects against embryotoxicity and teratogenicity of Cd.     

Palatal Slit and Cleft Palate in Rats Treated with Glucocorticoids II. Comparative Teratogenicity of Prednisolone, Triamcinolone Acetonide and Hydrocortisone

The induction of abnormal palatal development by three glucocorticoids; prednisolone, triamcinolone acetonide and hydrocortisone was evaluated in rat fetuses. Pregnant rats were injected subcutaneously with either prednisolone (12.5-100 mg/kg/day), triamcinolone acetonide (0.25-2 mg/kg/day) or hydrocortisone (100 mg/kg/day) on days 14 and 15 of gestation. These females were humanely killed on day 20 of gestation and viable fetuses were inspected for their palatal abnormalities. The frequencies of cleft palate were significantly higher in the group treated with 100 mg/kg/day prednisolone (10.6%), and in the groups treated with 0.5, 1 and 2 mg/kg/day triamcinolone acetonide (8.6%, 26.1% and 58.3%, respectively) than the control frequency of 0%. Triamcinolone acetonide was 70 times as potent as prednisolone in inducing palatal slit, with ED₅₀ value of 1.0 mg/kg/day and 70 mg/kg/day, respectively. Hydrocortisone showed no potentiality for the induction of cleft palate and palatal slit. Other developmental abnormalities including omphalocele and general edema, late resorption, and growth retardation were induced by triamcinolone acetonide and prednisolone. These findings indicate that triamcinolone acetonide has a significantly higher potentiality for the induction of palatal slit in rats, as well as in mice, compared to prednisolone and hydrocortisone.