Deletion 15q24-26 in prenatally detected diaphragmatic hernia: increasing evidence of a candidate region for diaphragmatic development.

Survival of children with congenital diaphragmatic hernia (CDH) is mainly dependent on the extent of lung hypoplasia and the presence of additional congenital anomalies or chromosomal aberrations. A chromosomal deletion 15q25-q26.2 in a fetus with prenatally diagnosed CDH and growth retardation is reported. Despite optimal pre- and neonatal management the baby died shortly after birth. There is increasing evidence that the long arm of chromosome 15, and especially the region 15q24 to 15q26, plays a crucial role in the development of the diaphragm. The finding of a deletion within 15q24-26 in a fetus with CDH has to be considered a predictor of poor prognosis. It is of utmost interest for proper parental counselling to search in fetuses with CDH for subtle chromosomal lesions paying special attention to chromosome 15q.

Prenatal diagnosis of heterokaryotypic mosaic twins discordant for fetal sex.

The presence of a monozygotic twin gestation with discordant sex of the twins is a very rare constellation, which is referred to as heterokaryotypic monozygotic pregnancy. This constellation can develop either due to a chromosomal aberration after twinning or is - as in the following case - due to a mitotic error before twinning and an unequal distribution of mosaicism in both embryos. So far the diagnosis of heterokaryotypic monozygotic pregnancy has always been made postnatally, with only one exception (Gonsoulin et al., 1990). In this case we suspected the presence of monozygotic twins ultrasonically because of the chorionic and amniotic membrane characteristics. Surprisingly the sex of the fetuses was discrepant. As one of them had hydrops and a structural heart defect, we carried out an amniocentesis, which revealed mosaicism [45,X/46,X,i(Y)(p10)] of both fetuses. The female fetus with a predominant 45,X set of chromosomes and the typical intrauterine signs of the Ullrich-Turner syndrome (massive hygroma colli, hydrops fetalis and multiple cardiac defects) died during the 25th week of gestation due to cardiac decompensation. The other fetus appeared to be male with a predominance of a 46,X,i(Y)(p10) set of chromosomes and was born a few days after the intrauterine death of the hydropic fetus. In conclusion, our observation shows that ultrasonic evidence of discordant fetal sex in twins does not necessarily exclude monozygosity.

All-artery multigraft coronary artery bypass grafting with only internal thoracic arteries possible and safe: a randomized trial.

BACKGROUND: The internal thoracic artery (ITA) bypass to the left anterior descending coronary artery is of proven benefit in multigraft coronary artery bypass. Total ITA grafts, if reoperation is averted by avoiding saphenous vein grafts (SVGs), are attractive. The safety of the total ITA graft operation (all-ITA) is a concern. METHODS: A randomized trial of multiple-ITA bypass graftings with the use of bilateral sequential ITA without SVGs was performed. Control patients received 1 ITA plus SVG. Inclusion criteria were those used in the Coronary Artery Surgery Study, extended to age 76 years, and any angina class, except emergent. One hundred sixty-two patients were randomized (81 patients per group) from January 1, 1990, to December 31, 1994. RESULTS: Baseline traits were similar as were cross-clamp times, pump times, and number of arteries bypassed (average, 4.3 arteries). Patients who received multiple ITA grafts had no myocardial infarctions, per reference laboratory. One patient died, and 2 patients returned for bleeding. The ITA-SVG group had similar results. The all-ITA group experienced successful completion in 93% of cases. Complications did not differ from control patients. CONCLUSIONS: Early and 5-year outcomes were not different between the all-ITA group and the ITA with SVGs group. We believe experienced surgeons can safely extend the ITA to multibypass coronary artery bypass without use of SVG to achieve an all-ITA operation.

A laboratory for testing the interoperability of telehealth systems.

Interoperability allows telehealth equipment to interact to achieve predictable results. To address the need for telehealth interoperability, the Alberta Research Council has been working with the Alberta Health and Wellness organization in Canada, and others, to create guidelines and a facility for testing telehealth equipment for compliance with technical interoperability standards. The laboratory consists of two rooms (7 m x 7 m) in a new building. The rooms are wired with easy-to-configure copper and fibre networks for telephone, Switch-56, ISDN, ATM, wireless and satellite services. One room specializes in teleconsultation and tele-education, while the other has facilities for teleradiology and telemonitoring. The rooms are interconnected in order to perform interoperability tests between realtime and store-and-forward equipment. The laboratory was piloted in the summer of 1999.

Effect of Florida's graduated licensing program on the crash rate of teenage drivers.

On 1 July 1996, Florida instituted a graduated licensing program for drivers younger than age 18. For the first 3 months, holders of learner's licenses are not allowed to drive at all between 19:00 and 06:00 h; thereafter, they may drive until 22:00 h. Learner's licenses must be held for 6 months prior to eligibility for the intermediate license. Sixteen-year-old intermediate license holders are not permitted to drive unsupervised from 23:00 to 06:00 h, 17 year-olds from 01:00 to 06:00 h. All drivers younger than 18 have strict limits on the number of traffic violations they can accumulate and, effective 1 January 1997, all drivers younger than 21 are subject to a zero tolerance law for drinking and driving. Florida crash data for 1995-1997 were obtained and compared with similar data from Alabama, a state that borders Florida but does not have graduated licensing. For 15, 16, and 17 year-olds combined, there was a 9% reduction in the fatal and injury crash involvement rate in Florida during 1997, the first full year of graduated licensing, compared with 1995. On a percentage basis, crashes declined most among 15 year-olds, followed by 16 year-olds and then 17 year-olds. Reductions were not seen among Alabama teenagers nor among 18 year-olds in Florida.

[Diagnosis of aneuploidy with fluorescence in situ hybridization (FISH); value in pregnancies with increased risk for chromosome aberrations]. / Aneuploidiediagnostik mittels Fluoreszenz-in-situ-Hybridisierung (FISH); Stellenwert bei Schwangerschaften mit erhöhtem Risiko für Chromosomenaberrationen.

BACKGROUND: In the case of abnormal ultrasound findings, abnormal serum-screening and age-risk in advanced pregnancy a rapid diagnosis or exclusion of a chromosomal aneuploidy of the fetus is of great value for the clinical management. With fluorescence in situ hybridization (FISH) on uncultured amniotic fluid cells the detection of the most common aneuploidies, which account for about 2/3 of all chromosomal aberrations [1], is possible within 24 hours. The aim was to evaluate if the FISH-technique in combination with karyotyping after cell culturing could replace other methods like diagnostics form umbilical cord blood or placental biopsies. MATERIALS AND METHODS: For the FISH assays commercially available directly with fluorochromes labelled DNA-probes (Vysis, Stuttgart) were used. FISH assays were performed on amniotic fluid samples from pregnancies at risk for fetal chromosome aberrations parallel to standard cytogenetic analysis. The method was performed on 230 samples of amniotic fluid. We tried to optimize the method concerning preparation of the cell material, the denaturation- and hybridization-steps and well as stringency of post-hybridization washes. RESULTS: All trisomies 13, 18, 21 and the sex chromosome aneuploidies (n = 34) which were diagnosed by conventional cytogenetics were identified correctly by FISH analysis with the exception of one case of trisomy 21 mosaicism, in which hybridization failed. As structural chromosome aberrations and mosaicisms cannot be detected with this method, six additional chromosome aberrations were identified exclusively by cytogenetic analysis. The mean frequency of nuclei with abnormal signal pattern in the aneuploid cases was 89%. A minimum of 50 nuclei for each DNA-probe could be counted in 86% of the samples. The results of 12 cases were classified as uninformative, because only less than 15 hybridized nuclei or no hybridization signals could be scored. Maternal contamination was found in 17.4% of the samples. CONCLUSIONS: In clinical cases with a high risk for an abnormal fetal karyotype and the need of quick clinical consequences, methods which make possible a karyotyping within shortest time should be preferred to amniocentesis and FISH-analysis, because Chromosomal mosaicism and structural aberrations, which represent up to 20% of all chromosomal abnormalities in this group, cannot be detected, uninformative cases can occur in up to 15% of all investigated samples and There is a risk for false-negative results through contamination of the sample with cells of maternal origin. In comparison with methods which permit rapid karyotyping from umbilical cord blood or placental biopsies, a delay in the diagnostic procedure has to be accepted, when the result of the FISH-analysis has to be confirmed by cell culturing and standard cytogenetic analysis.

Prevalence and characteristics of red light running crashes in the United States.

About 40% of motor vehicle crashes occur at intersections. In recent years, the number of crashes at traffic signals has increased considerably. A major cause of such crashes is drivers disregarding traffic signals. Despite concerns about the frequent occurrence of red light violations and the significant crash consequences, relatively little is known about the overall prevalence and characteristics of red light running crashes. The present study examines the prevalence of red light running crashes on a national basis and identifies the characteristics of such crashes and the drivers involved. Cities with especially high rates of fatal red light running crashes are identified. Countermeasures to reduce red light running crashes based on collision patterns and characteristics of drivers involved are discussed. It was estimated that about 260000 red light running crashes occur annually in the United States, of which approximately 750 result in fatalities. Comparisons were made between red light running drivers and drivers deemed not to have run red lights in these same crashes. As a group, red light runners were more likely than other drivers to be younger than age 30, male, have prior moving violations and convictions for driving while intoxicated, have invalid driver's licenses, and have consumed alcohol prior to the crash. Comparisons also were made between characteristics of red light runners involved in daytime and nighttime crashes. Nighttime red light runners were more likely than daytime runners to be young, male, and have more deviant characteristics, 53% having high blood alcohol concentrations.

Further observations of true mosaic trisomy 17 ascertained in amniotic fluid cell cultures.

Three new cases of true mosaic trisomy 17 (MT17) were diagnosed in amniotic fluid cells. Postnatal chromosome analysis from lymphocytes did not confirm the trisomic cell line, and follow-up studies showed normal psycho-motor development of the children, in one case up to the age of 4 1/2 years. We suggest that there are similarities between MT17 and MT20, in which the majority of pregnancies result in deliveries of healthy babies.

Tumor necrosis factor-alpha and interferon-gamma, but not HTLV-I tax, are likely factors in the epidermotropism of cutaneous T-cell lymphoma via induction of interferon-inducible protein-10.

We have previously shown that Interferon-Inducible Protein-10 (IP-10), a cytokine chemotactic for CD4-positive lymphocytes, is overexpressed by lesional epidermal keratinocytes and probably accounts for the epidermotropism of cutaneous T-cell lymphoma (CTCL). The tax gene of human T-lymphotropic virus-I (HTLV-I) immortalizes CD4-positive lymphocytes, induces IFN-gamma, and has been detected in patients with classical CTCL who are seronegative for HTLV-I. TNF-alpha is synergistic with IFN-gamma for the induction of IP-10. We therefore decided to define the presence of tax, IFN-gamma, TNF-alpha, and IP-10 in lesions of 19 adults with classical CTCL who were seronegative for HTLV-I. Lesional mRNAs for actin, TNF-alpha, IFN-gamma, and tax were detected by reverse-transcriptase polymerase chain reaction (RT-PCR) amplification. In addition IP-10, TNF-alpha, and IFN-gamma were detected and localized with immunocytochemistry of frozen sections. In agreement with previous observations IP-10 was overexpressed in lesional keratinocytes of all 19 patients. By RT-PCR, mRNA for IFN-gamma was detected in lesions of 8, and for TNF-alpha in lesions of 13 patients. By immunocytochemistry, TNF-alpha was expressed by lesional keratinocytes in 10 of 13 tested patients, whereas IFN-gamma was focally expressed by lesional lymphocytes and faintly by lesional keratinocytes in 9 of 13 tested patients. tax mRNA was not detected in lesions of any patient, but was easily detectable in cutaneous lesions or peripheral blood of control patients who were seropositive for HTLV-I. We conclude that TNF-alpha and IFN-gamma may cause epidermotropism by inducing IP-10. However, the tax gene of HTLV-I does not appear to be involved in the pathogenesis of classical CTCL.

Decreased placental and umbilical cord glycogen levels associated with meconium-stained amniotic fluid.

Most frequently, placental glycogen has been studied as an index of fetal nutrition. There are no published studies of placental glycogen as an index of fetal stress. In this study of 1573 samples from 71 placentae, glycogen levels in the placental disk, fetal membranes and umbilical cord of normal uncomplicated pregnancies were compared with those in complicated pregnancies. The complicated pregnancies included preterm delivery, hypertensive disorders, inadequate prenatal care, substance abuse, maternal fever or infection, obesity, diabetes mellitus, premature rupture of membranes, intrauterine growth retardation, sickle cell trait, and acute meconium staining of amniotic fluid at delivery. The data showed that the only significant differences were in the subgroup complicated by meconium-stained amniotic fluid in which the placental disks and umbilical cords had significantly lower (P=0.0006) glycogen levels. This finding suggests a relatively specific association. It is interesting to speculate that the passage of meconium with its vasoconstrictive effect increases utilization of local glycogen stores, decreases local glycogen reserves needed for the work of further vasoconstriction, and, in the event of subsequent acute stress, impairs vascular perfusion of tissues. In this way, meconium could predispose the infant to asphyxia.

Interstitial deletion del(3)(p12p21) in a malformed child subsequent to paternal paracentric insertion (or intraarm shift) 46,XY, ins(3)(p24.1p12.1p21.31).

We report on a malformed stillborn with deletion 3p subsequent to direct paracentric insertion (intraarm shift) in the normal father which had been first mistaken for paracentric inversion. The corrected diagnosis was supported by FISH of mapped markers on metaphase chromosomes. In addition we looked for recombinants in sperm. This observation reminds similar cases that had been considered exceptions to the expected meiotic recombination of paracentric inversions and points to a cytogenetic pitfall. Published deletions and paracentric inversions in 3p are briefly quoted.

Fatal crash risk for older drivers at intersections.

Risk of fatal crash involvement was calculated for older drivers relative to drivers aged 40-49 in the United States during the years 1994-1995. The results indicated that drivers ages 65-69 were 2.26 times more at risk for multiple-vehicle involvements at intersections compared with 1.29 times more at risk in all other situations. The comparable figures for drivers aged 85 and older were 10.62 for multiple-vehicle involvements at intersections compared with 3.74 for all other situations. The relative crash risk was particularly high for older drivers at uncontrolled and stop sign-controlled locations; when traveling straight or when just starting to enter the intersection; and when the specific behavioral error in the crash was failure to yield. Countermeasures will likely involve reducing or simplifying the need to detect and evaluate moving traffic coming from the left and right when at intersections. This can be accomplished by traffic signals with protected left turns, four-way stop signs, and one-way streets. Whereas such devices involve significant cost in terms of dollars and travel delay, their cost-effectiveness may have to be revisited as the United States population continues to age.

Epidermolytic acanthomas: clinical characteristics and immunohistochemical features.

Epidermolytic hyperkeratosis in bullous congenital ichthyosiform erythroderma results from mutations in the K1 and K10 genes. Epidermolytic acanthomas are solitary or multiple lesions with microscopic features that are identical to those in bullous congenital ichthyosiform erythroderma. In this study, the clinical and epidemiologic characteristics of epidermolytic acanthomas were summarized, and the expression of keratins (using antibodies to K1, K6, K10, K14, K16, and K19) in five solitary epidermolytic acanthomas was determined using immunohistochemistry techniques. The intensity of staining for K1 and K10 was (a) less in the altered granular layer, as compared to the adjacent nonaltered granular layer of the lesional skin, and (b) less in the lesional skin as compared to the perilesional, histologically normal-appearing skin. Expression of K6 and K16 was noted not only in the basal layer and suprabasal layers of the lesions, but also in the corresponding layers of the adjacent normal skin. Staining for K14 was also observed in the basal layers and suprabasal layers of the lesional and adjacent normal epidermis; within the lesional and perilesional normal skin, the intensity of positive staining for K14 was greater in the basal layers than in the suprabasal layers of the epidermis. The specimens did not stain for K19. In conclusion, using immunohistochemistry techniques on solitary epidermolytic acanthomas, we were able to demonstrate (1) an abnormality in K1 and K10 expression in the lesional skin as compared to the adjacent, histologically normal-appearing skin and (b) the expression of hyperproliferative keratins not only with the lesional skin, but also in the perilesional normal skin. We hypothesize that the pathogenesis of epidermolytic hyperkeratosis in lesions of solitary epidermolytic acanthomas results from mutations in the K1 and K10 genes.

Arrest of mycobacterial phagosome maturation is caused by a block in vesicle fusion between stages controlled by rab5 and rab7.

Mycobacterium tuberculosis and the closely related organism Mycobacterium bovis can survive and replicate inside macrophages. Intracellular survival is at least in part attributed to the failure of mycobacterial phagosomes to undergo fusion with lysosomes. The transformation of phagosomes into phagolysosomes involves gradual acquisition of markers from the endosomal compartment. Members of the rab family of small GTPases which confer fusion competence in the endocytic pathway are exchanged sequentially onto the phagosomal membranes in the course of their maturation. To identify the step at which the fusion capability of phagosomes containing mycobacteria is compromised, we purified green fluorescent protein-labeled M. bovis BCG phagosomal compartments (MPC) and compared GTP-binding protein profiles of these vesicles with latex bead phagosomal compartments (LBC). We report that the MPC do not acquire rab7, specific for late endosomes, even 7 days postinfection, whereas this GTP-binding protein is present on the LBC within hours after phagocytosis. By contrast, rab5 is retained and enriched with time on the MPC, suggesting fusion competence with an early endosomal compartment. Prior infection of macrophages with M. bovis BCG also affected the dynamics of rab5 and rab7 acquisition by subsequently formed LBC. Selective exclusion of rab7, coupled with the retention of rab5 on the mycobacterial phagosome, may allow organisms from the M. tuberculosis complex to avert the usual physiological destination of phagocytosed material.

Prenatal detection of double aneuploidy trisomy 10/monosomy X in a liveborn twin with exclusively monosomy X in blood.

Both double aneuploidy and trisomy 10 are rare chromosome findings. All five published cases of trisomy 10 in liveborns were found to be mosaic with an euploid cell line. In a liveborn female twin, double aneuploidy mosaicism 47,XX, + 10/45,X was detected prenatally by amniocentesis performed because of severe intrauterine growth retardation and malformations. Chromosome analysis from neonatal lymphocyte cultures revealed exclusively the 45,X cell line. Double aneuploidy mosaicism trisomy 10/monosomy X was confirmed from skin fibroblasts. The child died at the age of 7 weeks. This is the first reported case of double aneuploidy involving trisomy 10, and the first case of trisomy 10 without a normal cell line in a liveborn. Prenatal diagnosis of trisomy 10 in a liveborn has not been published so far. The case illustrates that in specific cases amniotic fluid cells may reflect the karyotype of the fetus better than blood.

Interferon-inducible protein 10 as a possible factor in the pathogenesis of cutaneous T-cell lymphomas.

Human IFN-gamma-inducible protein 10 (IP-10), a C-X-C chemokine secreted by IFN-gamma-stimulated keratinocytes, is chemotactic for normal CD4-positive lymphocytes and inhibits the proliferation of early subsets of normal and of leukemic hemopoietic progenitors. Cutaneous T-cell lymphoma (CTCL) is an indolent lymphoproliferative disorder of CD4-positive lymphocytes that remain confined to the skin for many years before visceral dissemination. Because IFN-gamma mRNA was detected in the epidermis of CTCL lesions, we decided to investigate the role of IP-10 in the epidermotropism of CTCL by determining its expression in normal skin and in CTCL lesions. Using purified recombinant IP-10 (rIP-10) or a recombinant fusion protein between IP-10 and the straight phi10 protein of phage T7, we generated rabbit antisera that recognized and neutralized rIP-10. Immunoperoxidase staining of normal epidermis demonstrated that IP-10 was expressed by basal keratinocytes but not by the more differentiated cells. In the often hyperplastic epidermis overlying CTCL lesions, IP-10 immunostaining was enhanced compared to normal skin and extended to the suprabasal keratinocytes in 28 of 29 patients for a frequency of 97% and a 95% confidence interval of 82-100%. However, IP-10 was detectable in the dermal or epidermal lymphoid infiltrates in only 3 of 29 patients (10%; 95% confidence interval, 2-29%). Skin clinically free of CTCL demonstrated normal IP-10 immunostaining. In one patient who had matching biopsies performed before and after treatment, IP-10 was overexpressed before treatment but was normally expressed at remission. The in vitro proliferation of primary normal human keratinocytes was inhibited in a dose-dependent manner by rIP-10. These results suggest that IP-10 plays a role in the epidermotropism of CTCL. Additional work is needed to determine whether IP-10 stimulates or inhibits CTCL proliferation. A better understanding of the growth controls operating in CTCL may be useful in the development of curative strategies for this disorder.

Multiple (up to seven) different accessory small marker chromosomes: prenatal diagnosis and follow-up.

We report on the prenatal discovery of 3 up to 7 accessory small marker chromosomes per cell with postnatal confirmation in various tissues. By FISH it could be shown that every marker had a different origin.