Nagato Natsume

Mutations in BMP4 Are Associated with Subepithelial, Microform, and Overt Cleft Lip

Cleft lip with or without cleft palate (CL/P) is a complex trait with evidence that the clinical spectrum includes both microform and subepithelial lip defects. We identified missense and nonsense mutations in the BMP4 gene in 1 of 30 cases of microform clefts, 2 of 87 cases with subepithelial defects in the orbicularis oris muscle (OOM), 5 of 968 cases of overt CL/P, and 0 of 529 controls. These results provide confirmation that microforms and subepithelial OOM defects are part of the spectrum of CL/P and should be considered during clinical evaluation of families with clefts. Furthermore, we suggest a role for BMP4 in wound healing.

In a Vietnamese population, MSX1 variants contribute to cleft lip and palate.

Purpose: To identify causes of nonsyndromic cleft lip and palate in a Vietnamese population.Methods: In this study, 175 families with at least one case of cleft lip and/or palate were studied using the candidate genes TGFA, MSX1, and TGFB3.Results: Transmission distortion for alleles of MSX1 were demonstrated for the whole population and two missense mutations were identified, including one (P147Q) that is found in approximately 2% of the population. The P147Q appears to arise from a founder individual based on shared haplotypes in unrelated families.Conclusions: MSX1 contributes to nonsyndromic clefting in a Vietnamese population, and consistent with other studies, identifiable mutations in this gene cause about 2% of cases of nonsyndromic clefting.

Three-dimensional analysis of facial morphology using moire' 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.

The prevalence of cleft lip and palate 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).

Replication of genome wide association identified candidate genes confirm the role of common and rare variants in PAX7 and VAX1 in the etiology of nonsyndromic CL(P).

Following recent genome wide association studies (GWAS), significant genetic associations have been identified for several genes with nonsyndromic cleft lip with or without cleft palate (CL(P)). To replicate two of these GWAS signals, we investigated the role of common and rare variants in the PAX7 and VAX1 genes. TaqMan genotyping was carried out for SNPs in VAX1 and PAX7 and transmission disequilibrium test (TDT) was performed to test for linkage and association in each population. Direct sequencing in and around the PAX7 and VAX1 genes in 1,326 individuals of European and Asian ancestry was done. The TDT analysis showed strong associations with markers in VAX1 (rs7078160, P = 2.7E−06 and rs475202, P = 0.0002) in a combined sample of Mongolian and Japanese CL(P) case–parent triads. Analyses using parent‐of‐origin effects showed significant excess transmission of the minor allele from both parents with the effect in the mothers (P = 6.5E−05, OR (transmission) = 1.91) more striking than in the fathers (P = 0.004, OR (transmission) = 1.67) for VAX1 marker rs7078160 in the combined Mongolian and Japanese samples when all cleft types were combined. The rs6659735 trinucleotide marker in PAX7 was significantly associated with all the US cleft groups combined (P = 0.007 in all clefts and P = 0.02 in CL(P)). Eight rare missense mutations found in PAX7 and two rare missense mutations in VAX1. Our study replicated previous GWAS findings for markers in VAX1 in the Asian population, and identified rare variants in PAX7 and VAX1 that may contribute to the etiology of CL(P). Determining the role of rare variants clearly warrants further investigation.

Cellular localization and functional characterization of the equilibrative nucleoside transporters of antitumor nucleosides

Nucleoside transporters play an important role in the disposition of nucleosides and their analogs. To elucidate the relationship between chemosensitivity to antitumor nucleosides and the functional expression of equilibrative nucleoside transporters (ENT), we established stable cell lines of human fibrosarcoma HT‐1080 and gastric carcinoma TMK‐1 that constitutively overexpressed green fluorescent protein‐tagged hENT1, hENT2, hENT3 and hENT4. Both hENT1 and hENT2 were predictably localized to the plasma membrane, whereas hENT3 and hENT4 were localized to the intracellular organelles. The chemosensitivity of TMK‐1 cells expressing hENT1 and hENT2 to cytarabine and 1‐(3‐C‐ethynyl‐β‐d‐ribopentofuranosyl) cytosine increased markedly in comparison to that of mock cells. However, no remarkable changes in sensitivity to antitumor nucleosides were observed in cell lines that expressed both hENT3 and hENT4. These data suggest that hENT3 and hENT4, which are mainly located in the intracellular organelles, are not prominent nucleoside transporters like hENT1 and hENT2, which are responsible for antitumor nucleoside uptake. (Cancer Sci 2007; 98: 1633–1637)

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.

Mutations in Extracellular Matrix Genes NID1 and LAMC1 Cause Autosomal Dominant Dandy–Walker Malformation and Occipital Cephaloceles

We performed whole‐exome sequencing of a family with autosomal dominant Dandy–Walker malformation and occipital cephaloceles and detected a mutation in the extracellular matrix (ECM) protein‐encoding gene NID1. In a second family, protein interaction network analysis identified a mutation in LAMC1, which encodes a NID1‐binding partner. Structural modeling of the NID1–LAMC1 complex demonstrated that each mutation disrupts the interaction. These findings implicate the ECM in the pathogenesis of Dandy–Walker spectrum disorders.

A Mutation in RYK Is a Genetic Factor for Nonsyndromic Cleft Lip and Palate

Objective The RYK, EPHB2, and EPHB3 genes are attractive candidates for cleft lip and/or palate and cleft palate only pathogenesis. Both the Ryk-deficient mouse and Ephb2/Ephb3 (genes for interaction molecules with RYK) double-mutant mouse show cleft palate. Setting Mutation searches for RYK, EPHB2, and EPHB3 were carried out in a large number of Japanese and Vietnamese patients with cleft lip and/or palate and cleft palate only. Case-control study and transmission disequilibrium tests were performed also, using three single nucleotide polymorphisms within a linkage disequilibrium block in RYK. Seven haplotypes were constructed from the single nucleotide polymorphisms. Results A missense mutation, 1355G>A (Y452C), in RYK was identified in one Vietnamese patient with cleft lip and/or palate. This mutation was not found among 1646 Vietnamese, Japanese, and Caucasians, including 354 cleft lip and/ or palate and cleft palate only patients. Colony formation assay using NIH3T3 cells transfected with mutant cDNA revealed that mutant RYK had significantly reduced protein activity, compared with those with wild-type RYK, implying that the transformation ability of RYK is depleted by this mutation. Although a case-control study and transmission disequilibrium tests on three individual single nucleotide polymorphisms provided no evidence for association with oral clefts, a case-control study on one rare haplotype suggested a positive association in Japanese patients with cleft lip and/or palate and cleft palate only. No mutations in EPHB2 and EPHB3 were found in any patients examined. Conclusion The findings suggested that a missense mutation, 1355G>A, and one rare single nucleotide polymorphisms haplotype may play a role in the development of cleft lip and/or palate in the Vietnamese, and cleft lip and/ or palate and cleft palate only in the Japanese.

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.