D20S16 is a complex interspersed repeated sequence: genetic and physical analysis of the locus.

The genomic structure of the D20S16 locus has been evaluated using genetic and physical methods. D20S16, originally detected with the probe CRI-L1214, is a highly informative, complex restriction fragment length polymorphism consisting of two separate allelic systems. The allelic systems have the characteristics of conventional VNTR polymorphisms and are separated by recombination (theta = 0.02, Zmax = 74.82), as demonstrated in family studies. Most of these recombination events are meiotic crossovers and are maternal in origin, but two, including deletion of the locus in a cell line from a CEPH family member, occur without evidence for exchange of flanking markers. DNA sequence analysis suggests that the basis of the polymorphism is variable numbers of a 98-bp sequence tandemly repeated with 87 to 90% sequence similarity between repeats. The 98-bp repeat is a dimer of 49 bp sequence with 45 to 98% identity between the elements. In addition, nonpolymorphic genomic sequences adjacent to the polymorphic 98-bp repeat tracts are also repeated but are not polymorphic, i.e., show no individual to individual variation. Restriction enzyme mapping of cosmids containing the CRI-L1214 sequence suggests that there are multiple interspersed repeats of the CRI-L1214 sequence on chromosome 20. The results of dual-color fluorescence in situ hybridization experiments with interphase nuclei are also consistent with multiple repeats of an interspersed sequence on chromosome 20.

Identification of sperm and non-sperm male cells in cervicovaginal smears using fluorescence in situ hybridization: applications in alleged sexual assault cases.

The identification of spermatozoa or constituents of seminal fluid is critical in the evaluation of alleged sexual assault victims. However, failure to identify sperm and/or elevated levels of acid phosphatase can occur for a variety of reasons. Molecular techniques, such as molecular cytogenetic analysis offers new approaches to improve on the identification of male cells in alleged sexual assault cases. Fluorescence in situ hybridization (FISH) with a Y chromosome specific DNA probe was applied to archival cervicovaginal smears from 41 alleged sexual assault cases to identify Y-bearing (male) cells. FISH identified Y-bearing sperm and non-sperm cells in 78% of the cases previously confirmed to have sperm. FISH also identified Y-bearing non-sperm male cells in 39% of the cases in which cytology did not detect spermatozoa; in one of these instances, it also detected sperm. Cervicovaginal acid phosphatase levels, determined at the time of the cervicovaginal smears, were also compared with the presence or absence of Y-positive cells. Application of this technique can detect non-spermatozoic male cells in routine cervicovaginal smears of sexual assault victims.

Molecular cytogenetic analysis of a duplication Xp in a male: further delineation of a possible sex influencing region on the X chromosome.

We describe a male infant with severe mental retardation and autism with a duplication of the short arm of the X chromosome. Chromosome painting confirmed the origin of this X duplication. Molecular cytogenetic analysis with fluorescence in situ hybridization (FISH) identified one copy of the zinc finger protein on the X chromosome (ZFX) and two copies of the steroid sulfatase gene (STS), further delineating the breakpoints. Based on cytogenetic and molecular comparisons of cases from the literature of sex-reversal in dup(X),Y patients and our patient, we suggest that a possible secondary sex-influencing gene involved in the regulation of sex determination or testis morphogenesis is present at the distal Xp21.1 to p21.2 region.

Prenatal detection of cri du chat syndrome on uncultured amniocytes using fluorescence in situ hybridization (FISH).

Fluorescence in situ hybridization (FISH) with a chromosome-region-specific DNA probe was used prospectively on uncultured amniocyte interphase cells to detect an unbalanced chromosome abnormality that resulted in cri du chat or 5p- syndrome. Confirmation was performed by routine cytogenetics.

Rapid detection of aneuploidy in uncultured chorionic villus cells using fluorescence in situ hybridization.

Rapid detection of aneuploidy using chromosome-specific repetitive DNA probes and the potential diagnostic accuracy of fluorescence in situ hybridization (FISH) on interphase cells of chorionic villus samples (CVS) are presented. Analyses demonstrated the ability to correctly identify aneuploidy using FISH in uncultured CVS. Our preliminary investigation suggests that this technique offers a significant clinical potential to circumvent problems of culture, time, and cost in cytogenetic analysis.

Unbalanced mosaic karyotypes with different structural abnormalities involving a common chromosome region: report of two cases.

We report on 2 cases with different de novo unbalanced mosaic karyotypes in which each cell line had a different structural abnormality involving a common chromosome region: 46,XX,del(11)(q23.3)/46,XX.-11, + der(11)t(11;?)(q23.3;?) and 46,X,idic(Xq)/46,X,idic(Xq),-12, + der(12)t(X;12)(p11.2;p13.3). Molecular-cytogenetic analysis confirmed the origin of the derivative 12 chromosome in the latter. We present a literature review of reports with mosaic cell lines of structural chromosome abnormalities that share the same chromosome breakpoint.

A genetic map of chromosome 20q12-q13.1: multiple highly polymorphic microsatellite and RFLP markers linked to the maturity-onset diabetes of the young (MODY) locus.

Multiple highly polymorphic markers have been used to construct a genetic map of the q12-q13.1 region of chromosome 20 and to map the location of the maturity-onset diabetes of the young (MODY) locus. The genetic map encompasses 23 cM and includes 11 loci with PIC values > .50, seven of which have PICs > .70. New dinucleotide repeat polymorphisms associated with the D20S17, PPGB, and ADA loci have been identified and mapped. The dinucleotide repeat polymorphisms have increased the PIC of the ADA locus to .89 and, with an additional RFLP at the D20S17 locus, the PIC of the D20S17 locus to .88. The order of the D20S17 and ADA loci determined genetically (cen-ADA-D20S17-qter) was confirmed by multicolor fluorescence in situ hybridization. The previously unmapped PPGB marker is closely linked to D20S17, with a two-point lod score of 50.53 at theta = .005. These markers and dinucleotide repeat markers associated with the D20S43, D20S46, D20S55, D20S75, and PLC1 loci and RFLPs at the D20S16, D20S17, D20S22, and D20S33 have been used to map the MODY locus on chromosome 20 to a 13-cM (sex averaged) interval encompassing ADA, D20S17, PPGB, D20S16, and D20S75 on the long arm of chromosome 20 and to create a genetic framework for additional genetic and physical mapping studies of the region. With these multiple highly polymorphic loci, any MODY family of appropriate size can be tested for the chromosome 20 linkage.

Physical localization of chromosome 20 markers using somatic cell hybrid cell lines and fluorescence in situ hybridization.

A panel of somatic cell hybrid cell lines containing different parts of human chromosome 20 and fluorescence in situ hybridization have been used to physically localize markers to human chromosome 20. Through these complementary approaches and genetic linkage analysis, D20S16, which is closely linked to the maturity onset diabetes of the young (MODY) locus, was mapped to band 20q12 --> q13.1. The gene for growth hormone-releasing factor (GHRF) was physically mapped and reassigned to 20q11, suggesting that GHRF plays no direct role in MODY. In addition, the genes for the chromosome 20-linked glycogen phosphorylase (GYPB) and the bone morphogenetic protein (BMP2A) have been assigned to chromosome 20p, and the interleukin-6-dependent DNA-binding protein (TCF5) has been assigned to 20q12 --> q13 by hybridization to genomic DNA from the panel of somatic cell hybrid cell lines. These approaches are useful for rapid localization of candidate genes for MODY and other DNA markers mapped to chromosome 20.

Molecular cytogenetic analysis of a familial 8p23.1 deletion associated with minimal dysmorphic features, seizures, and mild mental retardation.

We report a family in which three members presented with minimal phenotypic abnormalities, normal intelligence to mild mental retardation, and a cytogenetically terminal chromosome deletion at band 8p23.1 Whole chromosomal painting with a chromosome 8-specific DNA library confirmed this familial chromosome abnormality as a deletion, while fluorescence in situ hybridization with telomeric probes demonstrated the presence of telomeres at the deletion site. Coagulation studies were additionally performed to evaluate the purported location of the coagulation factor VII regulator gene at 8p23.1. A review of the clinical findings of seven cases of del(8)(p23.1) is presented.

Diplopia following blepharoplasty.

The complication of extraocular muscle palsy following blepharoplasty is rare. In a review of 920 blepharoplasties at Manhattan Eye, Ear and Throat Hospital, three well-documented cases of diplopia following blepharoplasty could be found. Only one of these cases resolved within two months postoperatively. The explanation offered for this phenomenon is a Volkmann type contracture of the extraocular muscles following edema and hemorrhage into the muscle sheath.

OCTOPUS--a church-based sex education program for teens and parents.

OCTOPUS is the acronym for a rural, church-based sex education program for teens and parents. The tentacles symbolize the agencies and individuals involved in this multi-faceted community outreach program designed to promote "Open Communication Regarding Teenagers Or Parents Understanding of Sexuality." Its purpose was to establish a forum for family discussion within a church setting to enhance communication skills, convey factual information, and cultivate the development of a decision-making process to help parents help their teenagers acquire appropriate morals and values. The OCTOPUS program was a team effort comprised of nurses, health educators, and counselors with experience serving adolescents, ministers who sought to integrate religious views with sex education, and church and community members interested in improving dialogue between parents and teenagers about sexuality. The team developed a comprehensive yet flexible program that could be modified to meet each church's preferences. Generally, the topics were arranged into four two-hour sessions. Presentation methods included a combination of lectures, films, and pamphlets. Large and small group discussions were used for clarification and communication skills development. While the results of this program were not quantifiable, feedback from four churches and one-hundred participants has been highly favorable.

PIP: OCTOPUS is the acronym for a rural, church-based sex education program for teens and parents. The tentacles symbolize the agencies involved in this multi-faceted community outreach program designed to promote "Open Communication Regarding Teenagers Or Parents Understanding of Sexuality." Its purpose was to establish a forum for family discussion within a church setting to enhance communication skills, convey factual information, and cultivate the development of a decision making process to help parents help their teenagers acquire appropriate morals and values. The OCTOPUS program was a team effort using 1) nurses, health educators, and counselors with experience serving adolescents, 2) ministers who wanted to integrate religious views with sex education, and 3) church and community members interested in improving communication about sex between parents and teenagers. The team developed a comprehensive but flexible program that could be modified for each church. Generally, four two-hour sessions covered love and relationships, being an adolescent, decision making, reproduction and pregnancy, birth control and sexually transmitted diseases, the church's view, communication skills, and community resources. Presentation methods combined lectures, films, and pamphlets. Large and small group discussions were used for clarification and communication skills development. While the results of this program were not quantifiable, feedback from four churches and 100 participants has been highly favorable.