Shinya Matsuura

Characterization of a 5′-flanking region of the human liver/bone/kidney alkaline phosphatase gene: Two kinds of mRNA from a single gene

We have cloned the human liver/bone/kidney alkaline phosphatase (ALPL) gene using a liver-type ALPL cDNA as a probe. The gene is divided into 12 exons, and is likely to exist as a single copy in haploid genome. As compared with the gene isolated using a bone-type ALPL cDNA (Weiss et al., J. Biol. Chem. 263, 12002-12010, 1988), another leader exon specific for the liver-type ALPL mRNA was assigned about 3.4 kb upstream from exon 2 and the alternative splicing in the first exon was indicated. RNA blot analysis showed that three species of mRNA of 2.5, 4.1 and 4.7 kilobases were detected in liver and developmentally regulated.

Leukocyte adhesion deficiency: Identification of novel mutations in two japanese patients with a severe form

Leukocyte adhesion deficiency is a disorder with mutations of the gene for the beta subunit, a component common to three adhesion molecules; LFA-1, Mac-1 and p150,95. The molecular basis of the disorder was studied in two patients with its severe form. In the first patient, the mutant gene expressed an aberrant mRNA, 1.2 kb longer than usual, resulting from a G to A substitution at the splice donor site of a 1.2 kb intron. Several aberrantly spliced messages, arising from splicing at cryptic donor sites, were also identified. The beta subunit proteins deduced from the mRNA sequences lacked half the carboxyl terminal portion. In the second patient, the mutation was a G to A transition at nucleotide 454, which resulted in an Asp128 to Asn substitution of the beta subunit. The 128th Asp residue is located in a region crucial for the association with alpha subunits and strictly conserved among the integrin beta subunits so far analyzed.

Radiographic measurements of metacarpophalangeal lengths in Japanese children

SummaryThe lengths of 19 tubular bones of the hand were measured on radiographs of 1,585 Japanese children aged 0 to 17 years. The means and standard deviations related to age and sex were calculated and presented in tables. Using the standards, examples of the metacarpophalangeal pattern profile analysis are presented.

Inverted insertion (9)(q34.3q22.3q21.2) and its recombination product: Duplication 9q21.2q22.3

To the Editor: We read with interest a recent article in your journal by Dr. Narahara and his colleagues (1986) who described a kindred with two carriers of what they interpreted as an intrachromosomal direct shift, dir ins(9)(q34.3q22.1q31.3), and a now 9-yearold girl with a recombinant chromosome 9. It was deduced that the rec(9) in the girl resulted from crossing over at one of two loops formed during meiosis I in her carrier mother, and thus carried duplication of the 9q22.1---~q31.3 segment. The ABO locus was assigned to 9q31.3-~ qter in view of the fact that the girl was a recombinant for the locus. We studied another kindred in which an apparently identical ins(9) chromosome is segregating through five generations. Our kindred and the maternal grandfather of the proband in Naraharas kindred both live in Tokuyama, a city with a 113,000 population. It is thus likely, but yet to be proven, that the two kindreds are related with each other. Our interpretation of the ins(9) is different from Naraharas (Fig. l, upper row). It involves insertion of an inverted 9@1.2,q22.3 segment into 9q34.3. Pairing of the inv ins(9) chromosome with a normal chromosome 9 during meiosis I in a carrier individual would produce two loops, one involving the inverted q21.2--~ q22.3 segment and the other the @2.3--+q34.3 segment (Fig. l, lower row). Odd numbers of crossing over in the latter loop would produce a recombinant chromosome with duplication of the q21.2--, q22.3 segment and one with deficiency of the same segment (Fig. 2). Family studies in our kindred disclosed three ins(9) carriers and two individuals with dup 9q21.2--,q22.3 (Fig. 3). It was deduced that both l 1-2 and 11-3, and also either I-i or I-2, were carriers of the inv ins(9) chromosome. Thus, the trait was transmitted through at least five generations. V-4, the proband in our kindred, a 2 year 2 month-old girl with dup 9q21.2 ~-, q22.3, weighed 1,874 g at birth. She walked at 15 months but did not speak meaningful words at age 2 2/12 years. She measured 76.6 cm ( 3 . 2 SD) and weighed 8 kg ( 3 SD). She had ocular hypertelorism, a short nose with a depressed nasal tip, short neck, low-set, malformed ears, fifth finger clinodactyly, absence of bilateral palmar triradii C and distally placed axial triradii. Her bone age was correspondent to her chronological age. III-5, a maternal distant relative of the proband, also carries the dup(q) chromosome. She is now 45 years old and mentally retarded, but no other details are known to us. The proband in Naraharas kindred, a 5year-old girl with dup 9q, had a low birth weight, growth deficiency, ocular hypertelorism, and dermatoglyphic abnormalities. Her IQ was 92 and her bone age

Telomere association of human chromosomes induced by aphidicolin

Telomere associations were studied in metaphase chromosomes from 96-h cultures of peripheral blood lymphocytes of two healthy women, treated with 0.4 microM aphidicolin for the last 72 h. Telomere associations were encountered in 2.9% and 3.2% of the metaphases screened, whereas no such associations were encountered in 5-fluorodeoxyuridine-treated cultures. The chromosome arms involved in telomere associations were nonrandom: 1q, 2q, 3q, 6p and 16q were more frequently involved in the associations (P less than 0.01). Of the 51 combinations of telomere associations encountered, those occurring nonrandomly were 1q/2q, 2q/2q, 4q/4q, 6q/6q and 6p/6p associations.

Isolation of A Y chromosomal DNA sequence and its clinical application

SummaryA 4.6 kb long, Y-specific DNA fragment was isolated from a flow-sorted human Y chromosomal library, and its male specificity was confirmed by Southern blot analysis. The fragment, designated as pY-80, was proven with an in situ hybridization experiment to have originated from the Yp11.2-Ypter region. Its 2,808 bp section was sequenced. The polymerase chain reaction proceeded with oligonucleotides flanking a 666 bp PstI-EcoRI fragment of the sequence as primers and a male genomic DNA as a template, but not with a female genomic DNA. Preliminary tests of samples of various sources successfully detected the Y-specific fragment in male-derived samples, including mouth wash, single hair roots, urinary epithelial cells, dried blood spots and amniotic fluid cells.

Y-derived sequence detected in minute chromosomes by polymerase chain reaction and in situ hybridization

A 10‐year‐old girl and a 10‐month‐old girl, both with ambiguous genitalia, were found to have 45,X/46,X,mar and 45,X/46,X,r(?) mosaicism. The marker chromosomes in both girls were very small. Polymerase chain reaction, with synthetic oligonucleotide primers from Y‐specific DNA sequences pY‐80 and pY53.3 containing the sex‐determining region Y(SRY), proved the marker chromosomes to contain the Y short arm material. In situ hybridization with probe pY‐80 confirmed that the marker chromosomes included the Y short arms. These findings, together with ambiguous genitalia in the girls, indicate that the marker chromosomes include the testis‐determining factor gene.