A novel missense mutation in Van der Woude syndrome: usefulness of fingernail DNA for genetic analysis.

Van der Woude syndrome (VWS) is an autosomal-dominant oral facial disorder. To find a gene mutation in a Japanese family using fingernail DNA samples, we performed this study. We hypothesized that a gene mutation in IRF6 might be involved in VWS, and that fingernail DNA samples may be valuable for detecting such mutations. Linkage and haplotype analyses of the family mapped the disease locus to the 1q32-q41 region. Mutation analysis with an improved extraction method for fingernail DNA detected a novel missense mutation (1046A>T, E349V) in exon 7 of IRF6 in all the affected members of the family. Since the E349V change may disturb the hydrophobic core and affect regulatory activity of IRF6, it is most likely that the mutation is causative for VWS in this family. Fingernail DNA is thus useful for linkage and mutation analyses, since the fingernail can be easily obtained non-invasively, sent through the mail, and stored for a long period. We emphasize here the usefulness of fingernail DNA for the genetic analysis of a disease.

Complete sequencing shows a role for MSX1 in non-syndromic cleft lip and palate.

MSX1 has been proposed as a gene in which mutations may contribute to non-syndromic forms of cleft lip and/or cleft palate. Support for this comes from human linkage and linkage disequilibrium studies, chromosomal deletions resulting in haploinsufficiency, a large family with a stop codon mutation that includes clefting as a phenotype, and the Msx1 phenotype in a knockout mouse. This report describes a population based scan for mutations encompassing the sense and antisense transcribed sequence of MSX1 (two exons, one intron). We compare the completed genomic sequence of MSX1 to the mouse Msx1 sequence to identify non-coding homology regions, and sequence highly conserved elements. The samples studied were drawn from a panethnic collection including people of European, Asian, and native South American ancestry. The gene was sequenced in 917 people and potentially aetiological mutations were identified in 16. These included missense mutations in conserved amino acids and point mutations in conserved regions not identified in any of 500 controls sequenced. Five different missense mutations in seven unrelated subjects with clefting are described. Evolutionary sequence comparisons of all known Msx1 orthologues placed the amino acid substitutions in context. Four rare mutations were found in non-coding regions that are highly conserved and disrupt probable regulatory regions. In addition, a panel of 18 population specific polymorphic variants were identified that will be useful in future haplotype analyses of MSX1. MSX1 mutations are found in 2% of cases of clefting and should be considered for genetic counselling implications, particularly in those families in which autosomal dominant inheritance patterns or dental anomalies appear to be cosegregating with the clefting phenotype.

Analysis of transepidermal leukocyte chemotaxis in experimental dermatophytosis in guinea pigs.

In dermatophytosis, there is exocytosis of polymorphonuclear leukocytes (PMNs) toward the fungus-laden horny layer. To analyze this mechanism, we studied in vitro leukotactic properties of epidermal extracts prepared from lesions of experimental Trichophyton mentagrophytes infection in guinea pigs as well as fungus-derived chemotactic factors. Trichophyton mentagrophytes was found to release low-molecular-weight PMN chemotactic factors with its growth, like other microorganisms. Furthermore, it activated complement via the alternative pathway to produce complement-derived chemotactic factors in vitro. The epidermal extracts prepared from weakly inflammatory lesions of an early stage of T. mentagrophytes infection showed mild PMN chemotactic activity, which, as in those from irritated skin, was mostly due to the presence of low-molecular-weight chemotactic factors. After 8 days of infection, when prominent PMN migration took place together with the development of immune reactivity to fungus antigens, the epidermal extracts revealed strong leukotactic activity that showed a triphasic pattern by Sephadex G-75 chromatography similar to that observed in complement-activated serum. Since we could not demonstrate any deposition of immunoglobulins and complement on the fungal elements present in the horny layer, actual complement activation in vivo seems to occur after interaction of serum with soluble fungal components in the epidermis through both classic and alternative pathways. We think that the transepidermal migration of PMNs in dermatophytosis, together with contact sensitivity to fungal antigens, is responsible for induction of increased epidermopoiesis with resultant desquamation to eliminate the fungus-laden horny layer.

Correlation between the age of pinealectomy and the development of scoliosis in chickens.

STUDY DESIGN: Pinealectomy induces experimental scoliosis in chickens. This study analyzed the correlation between the age at which pinealectomy was performed and the development of scoliosis in chickens. OBJECTIVE: To investigate the differences in the rate or magnitude of scoliosis and the type of curvature in chickens pinealectomized at different times after hatching. SUMMARY OF BACKGROUND DATA: Scoliosis develops in almost all chickens pinealectomized within 3 days after hatching, but there are no data on whether the condition will develop in chickens pinealectomized earlier or later after hatching. METHODS: In this study, 106 female white leghorn chickens were divided into six groups: four pinealectomy groups (pinealectomy was performed 2, 4, 11, or 18 days after hatching in Groups P-2, P-4, P-11, and P-18, respectively), a control group (Group C), and a sham operation group (Group S). Ventrodorsal radiographs of the spine were taken at 4-week intervals until the age of 12 weeks. At 12 weeks, a 1-mL sample of blood was taken from the heart at the middle of the dark cycle, and the serum melatonin concentration was measured by radioimmunoassay. RESULTS: At the age of 12 weeks, scoliosis was present in 63.6% of the chickens in Group P-2, 72.7% in Group P-4, 81% in Group P-11, and 70% in Group P-18, and the Cobb angles in the scoliotic chickens averaged 32.6, 29.8, 23.8, and 22.3 degrees in the respective groups. There were no significant differences in the rate or magnitude of scoliosis and the type of curvature among the pinealectomy groups at the age of 12 weeks. At the age of 12 weeks, the serum melatonin levels at the middle of the dark cycle in the pinealectomized chickens were significantly lower than those of chickens in Groups C and S. However, there were no differences in the serum melatonin levels between scoliotic and nonscoliotic pinealectomized chickens. CONCLUSIONS: Findings from this study show that scoliosis develops in 60% to 80% of chickens pinealectomized within 18 days after hatching, and that scoliotic development is not influenced by the age at which pinealectomy is performed. However, this study suggests that melatonin plays a complicated role in spinal development, inasmuch as the serum melatonin levels after pinealectomy approximated zero. Yet scoliosis did not develop in all pinealectomized chickens.

Analysis of cartilage-derived retinoic acid-sensitive protein in cerebrospinal fluid From patients With spinal diseases.

STUDY DESIGN: The expression of cartilage-derived retinoic acid-sensitive protein (CD-RAP) was measured in cerebrospinal fluid from patients with spinal diseases. OBJECTIVES: To quantify the levels of CD-RAP in human cerebrospinal fluid and to clarify its character. SUMMARY OF BACKGROUND DATA: Cartilage-derived retinoic acid-sensitive protein is a newly discovered, secreted molecule that is expressed during the chondrogenesis phase of endochondral bone formation and in articular cartilage. In recent studies CD-RAP has been detected in the serum of patients with melanoma and breast cancer, and it has been used to monitor tumor activity. However, the function of CD-RAP is unknown, and the expression of CD-RAP in human cerebrospinal fluid has never been reported. METHODS: The concentration of CD-RAP in human cerebrospinal fluid was measured by enzyme-linked immunosorbent assay with antihuman CD-RAP antibodies. Cerebrospinal fluid samples were collected from two groups of patients. Group 1, the control group, consisted of 40 patients: 22 with trauma and 18 with gynecologic diseases. Group 2 consisted of 172 patients with spinal diseases: 5 with meningioma, 5 with neurinoma, 5 with arachnoid cyst, 30 with cervical spondylotic myelopathy, 35 with lumbar disc herniation, 56 with lumbar canal stenosis, and 36 with scoliosis. RESULTS: The concentration of CD-RAP in the control group was 16.5 +/- 8.3 ng/mL. The concentrations of CD-RAP in Group 2 were: 35.3 +/- 14.7 ng/mL in meningioma, 23.5 +/- 7.41 ng/mL in neurinoma, 26.0 +/- 22.2 ng/mL in arachnoid cyst, 41.7 +/- 22.3 ng/mL in cervical myelopathy, 27.8 +/- 14.7 ng/mL in lumbar disc herniation, 36.5 +/- 18.4 ng/mL in lumbar canal stenosis, and 13.4 +/- 7.48 ng/mL in scoliosis. The concentrations of CD-RAP in cervical myelopathy, lumbar canal stenosis, and lumbar disc herniation were significantly higher than in the control group (P < 0.001). CONCLUSIONS: The CD-RAP concentration was low in the control group, whereas it was significantly higher in spinal diseases that cause spinal stenosis. CD-RAP is expressed in cerebrospinal fluid as a result of damage to or stressing of neural structures and could be a marker for spinal diseases.

Incidence of cleft lip and cleft palate in 39,696 Japanese babies born during 1983.

To estimate the incidence of cleft lip and/or cleft palate among the Japanese, 39,696 Japanese babies born during the period from January 1, 1983 to December 31, 1983, were investigated. It was found that 65 babies (0.163%) had these abnormalities, and that the ratio of the birth of such babies was approximately one per 611 in the population. Of the 65 affected babies, 41.3% presented cleft lip (CL), 46.0% cleft lip and palate (CL/P), and 12.7% cleft palate (CP). These results suggest that the incidence of CL/P in the Japanese is higher than that observed among other races.

Teratogenesis of dexamethasone and preventive effect of vitamin B12.

Ddy mice which were treated with 0.32 mg of dexamethasone were used. Vitamin B12 was injected subcutaneously into the animals from the 10th day to the 13th day with a dosage of 0.1r, 1r, or 10r. The rate of incidence of cleft palate was decreased to 77.0% in the 0.1r group, 58.1% in the 1r group and 67.9% in the 10r group as compared to the control value of 88.5%.

Philtrum creation in secondary unilateral cleft lip repair.

A new method to create a philtrum in patients with unilateral cleft lip is presented. This modification was primarily used in secondary repair but can also be used in some primary repair cases. The results were satisfactory in most of the patients, at least at 2 years postoperatively.

Three-dimensional analysis of facial morphology using moiré stripes. Part I. Method.

A system is presented for analysing facial morphology using moiré stripes. Faces are photographed with a moiré camera, the moiré stripes are entered into a computer, and the computer automatically analyses the dimensions of the face. Subtle differences between the right and left sides of the face appear as asymmetries in the moiré pattern and are easily identified. The method is useful for analysing maxillofacial deformities, for treatment planning, and assessment of postoperative results.

Three-dimensional analysis of facial morphology using moiré stripes. Part II. Analysis of normal adults.

This paper presents a system for analysing facial morphology using moiré stripes in 60 normal Japanese high-school students. Analysis of the central region of the face revealed consistent differences between the sexes and between the right and left sides of the face. On the basis of these data, it is suggested that 1 mm difference should be considered permissible between the 2 sides of the upper lip in cases of lip repair in cleft lip and palate patients.

Modified technique in surgical correction of macrostomia.

A technique is presented to correct macrostomia with a simple straight line incision and incorporation of a small triangular flap to achieve proper positioning of the commissure with minimal visible scar.

Survey of patients with oblique facial clefts in Japan.

Oblique facial clefts constitute approximately 0.20% of all facial malformation cases in Japan and approximately 0.22% in other countries. In the present study, the proportion in our institute was approximately 0.21%, which is almost equal to that reported by WILSON et al. The ratio of male and female patients did not differ significantly from that in other countries.

Odontogenic infection pathway to the submandibular space: imaging assessment.

The aims of this study were to determine the pathways of odontogenic infection spread into the submandibular space and their relationship to the clinical symptoms. Computerized tomography (CT) and magnetic resonance (MR) images of 33 patients with submandibular involvement were analyzed. The spread of infection was evaluated by lateral asymmetry of the shape and density of the fascial spaces and tissues, and by obliteration of the interfascial fat spaces. Imaging findings were classified into three types: in 19 patients (57.6%), infection spread through the mylohyoid muscle or sublingual space (type I). In five patients (15.2%), infection spread through the bony structures of the mandible with periosteal reaction or perforation of the cortical plate (type II) and was associated with relatively mild symptoms. In four patients (12.1%), infection spread from the masticatory space (type III). Seven of 11 patients with dysphagia or fever showed submandibular involvement spreading into the parapharyngeal space. CT and MR imaging clearly demonstrated different pathways of the spread of odontogenic infection into the submandibular space, which influenced the manifestation of clinical symptoms.

Survey of congenital anomalies associated with cleft lip and/or palate in 701,181 Japanese people.

There have been many reports on congenital anomalies associated with cleft lip and/or palate (CL/CLP) in Japan. However, these reports included data only on patients who came to hospitals; thus the real situation regarding these anomalies remains unclear. Therefore, we surveyed newborns at all delivery facilities in the central area of Japan for the presence of these anomalies, following their progress for 12 consecutive years; at the end of that time, questionnaires were collected and analyzed. In this article, we describe our results.

The prevalence of cleft lip and plate in Japanese.

To determine the prevalence of cleft lip and palate (CL/P) among the Japanese, 43,821 babies born between 1 January, 1985, and 31 December, 1985, were investigated. Sixty-four infants (0.146%) were found to demonstrate these abnormalities, a birth prevalence rate of 1.46/1000. Among 326 infants in whom it was possible to classify the types of cleft, born between 1 January, 1981, and 31 December, 1985, there were 111 (34.0%) with cleft lip (CL), 154 (47.3%) with cleft lip and palate (CLP) and 61 (18.7%) with cleft palate (CP).

Maternal risk factors in cleft lip and palate: case control study.

Three hundred and six mothers who gave birth to babies with cleft lip, or palate, or both, were matched with 306 who gave birth to healthy babies in the same area during the same time period. Significantly more babies in the cleft group had a family history of clefts (48/306 compared with 7/306, P<0.0001). In the cases studied, combined cleft lip and palate was significantly more common among boys (82/157 compared with 57/149, P=0.02) and cleft palate alone among girls (48/149 compared with 22/157, P=0.0002). Significantly more mothers reported some sort of illness during early pregnancy (101/306 compared with 74/306, P=0.02). There were no differences between the groups as far as dietary preferences were concerned but during early pregnancy the mothers who gave birth to babies with defects tended to drink less alcohol (<1 unit/week) (236 compared with 199, P=0.001) and less coffee (<1 cup/week) (159/306 compared with 131, P=0.03). However, in each case similar proportions gave up once the pregnancy was confirmed. Large multicentre studies are required to confirm or refute these findings.

Incidence of cleft lip or palate in 303738 Japanese babies born between 1994 and 1995.

To investigate the incidence of cleft lip or palate or both (CLP) in Japan, 303738 babies born in 1532 institutions between 1994 and 1995 were examined and 437 (0.14%) were found to have abnormalities. Of these babies, 32.1% had cleft lip, 43.3% had cleft lip and palate, and 24.8% had cleft palate (Table 2). These results show that the incidence of cleft lip and palate has declined compared with the period from 1981 to 1982.