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Telomeres are nucleoprotein structures located at the termini of chromosomes that protect the chromosomes from fusion and degradation. Hepatocyte cell-cycle turnover may be a primary mechanism of telomere shortening in hepatitis C virus (HCV) infection, inducing fibrosis and cellular senescence. HCV infection has been recognized as potential cause of B-cell lymphoma and hepatocellular carcinoma. The present study sought to assess relative telomere length in leukocytes from patients with chronic HCV infection, patients after eradication of HCV infection (in remission), and healthy controls. A novel method of manual evaluation was applied. Leukocytes derived from 22 patients with chronic HCV infection and age- and sex-matched patients in remission and healthy control subjects were subjected to a fluorescence-in-situ protocol (DAKO) to determine telomere fluorescence intensity and number. The relative, manual, analysis of telomere length was validated against findings on applied spectral imaging (ASI) in a random sample of study and control subjects. Leukocytes from patients with chronic HCV infection had shorter telomeres than leukocytes from patients in remission and healthy controls. On statistical analysis, more cells with low signal intensity on telomere FISH had shorter telomeres whereas more cells with high signal intensity had longer telomeres. The findings were corroborated by the ASI telomere software. Telomere shortening in leukocytes from patients with active HCV infection is probably due to the lower overall telomere level rather than higher cell cycle turnover. Manual evaluation is an accurate and valid method of assessing relative telomere length between patients with chronic HCV infection and healthy subjects.

A case control study was performed to examine possible morbidity associated with Chlamydia trachomatis in 15 pregnant women with idiopathic preterm premature rupture of the membranes (PROM; group A), and in two control groups, 35 healthy preterm pregnant women (group B), and 43 healthy pregnant women at term (group C). Serum C. trachomatis IgG and IgA specific antibodies were determined using the single serovar (L2) inclusion immunoperoxidase assay. There were no significant differences in the prevalence rate of elevated levels of chlamydia IgG specific antibodies (titer greater than or equal to 1:128) between pregnant women suffering from idiopathic preterm PROM, as compared to healthy preterm and term pregnant women (20, 28 and 26%, respectively). Nor were there any significant differences in the prevalence rate of elevated levels of chlamydia IgA specific antibodies (titer greater than or equal to 1:16) between pregnant women with idiopathic preterm PROM, as compared to healthy preterm and term pregnant women (20, 20 and 17%, respectively). These findings do not support the assumption that C. trachomatis may play role in preterm PROM.

A study was undertaken to examine possible transplacental passage of Chlamydia trachomatis from pregnant women who were seropositive for chlamydia, but with no cervical C. trachomatis infection. Forty asymptomatic pregnant women, scheduled for diagnostic amniocentesis at 15–19 weeks of gestation, were tested for the presence of high serum IgA and IgG chlamydial specific antibodies and for cervical chlamydia infection. Five (12.5 %) had both high serum IgA and IgG antibody levels and 10 (25 %) had high serum IgG antibody levels. Overall, 15 (37.5%) had high serum chlamydia specific antibody levels (all were free of cervical chlamydial infection). The evaluation of the amniotic fluid specimens of these 15 seropositive pregnant women, who were free of cervical chlamydial infection, proved negative for direct C. trachomatis antigen detection and for chlamydial IgA and IgG specific antibodies. These negative results could be attributed to the lack of transplacental passage of C. trachomatis or to the antimicrobial activity of amniotic fluid against C. trachomatis, which has been previously described. The discrepancy between maternal infection and maternal serum antibody levels may suggest that the serologic test does not predict the presence of an antigen in the cervix.

Chronic myeloid leukemia (CML) is characterized by the presence of a BCR-ABL fusion gene, which is the result of a reciprocal translocation between chromosomes 9 and 22, and is cytogenetically visible as a shortened chromosome 22 (Philadelphia). Research during the past two decades has established that BCR-ABL is probably the pathogenetic pathway leading to CML, and that constitutive tyrosine kinase activity is central to BCR-ABL capacity to transform hematopoietic cells in vitro and in vivo. The tyrosine kinase inhibitor imatinib mesylate was introduced into the treatment regimen for CML in 1998. During the last few years, reports on chromosomal changes during imatinib treatment have been described. In this study, we evaluated the random aneuploidy rate with chromosomes 9 and 18 in bone marrow from treated and untreated patients. We found higher aneuploidy rates in both treated and untreated patients compared to the control group. In three patients who were treated with imatinib mesylate for more than 1.5 years, triploidy also appeared in some nuclei. To our knowledge, this is the first report on new chromosomal changes such as random aneuploidy and triploidy under imatinib treatment, but more studies are needed to investigate the long-term effect of the imatinib treatment on genetic instability.

Down syndrome (DS) is a multifactorial disorder with a high predisposition to leukemia and other malignancies. A change in the replication pattern from synchronous in normal genes to asynchronous in DS amniocytes has previously been reported. The objective of this study was to evaluate additional molecular cytogenetic factors which could re-emphasize the high correlation between DS cells and genetic instability. We found a higher rate of random aneuploidy in chromosomes 9 and 18 and a higher rate of asynchronous replication in the subtelomeric region or DS leukocytes than in cells from normal newborns. In addition, the telomere capture phenomenon was observed in the DS leukocytes but not in normal controls. The molecular cytogenetic factors observed in the DS individuals are known to correlate with genomic instability and with predisposition to cancer.

B-cell chronic lymphocytic leukemia (B-CLL) is the most common leukemia of adults in Western countries. The most frequent recurring chromosomal aberrations identified in B-CLL patients are trisomy 12 and deletions of 13q, 17p, and 11q. Cases with deletions of 11q and 17p have a poor prognosis, whereas cases with deletions in 13q have a favorable prognosis. It was previously shown that CLL patients with trisomy 12 and del(13)(q14) have a higher rate of asynchronous replication of normal structural genes when compared to those with normal karyotypes. We studied the replication pattern of the structural locus 21q22 and the imprinted gene SNRPN and its telomere (15qter) and the random aneuploidy of chromosomes 9 and 18 in CLL patients with trisomy 12 and deletions of 11q and 17p, and compared the results to those of CLL patients without these aberrations and to healthy controls. Random aneuploidy rate was higher in the group of patients with trisomy 12 as compared to all other groups. The replication pattern with higher asynchronous pattern was found in both aberration groups compared to the CLL patients without the aberrations and to the control group with involvement of 21q22 and 15qter, whereas the highest synchronous group was found in the 2 aberrations CLL patient groups compared to the other groups with the imprinted locus SNRPN. The existence and significance of chromosomal aberrations in CLL have a deleterious effect on the processes of cell cycle and gene replication and may have biological and prognostic implications.

A 33-year-old healthy woman, gravida 1 with twins pregnancy was admitted with mild preeclampsia and unusual hyponatremia which resolved promptly postpartum. This is the seventh reported case of hyponatremia complicating preeclampsia, four of the patients carried twins and four had nephrotic syndrome.

Telomeric regions of the human genome are of particular interest, because rearrangements of these regions are difficult to identify by conventional chromosome banding technology. With the advent of molecular cytogenetic techniques such as fluorescence in situ hybridization (FISH), it has been possible to investigate the terminus in cytogenetically visible terminal deletions and telomere rearrangements. We investigated telomere capture and aneuploidy rates in chronic lymphocytic leukemia (CLL) and chronic myeloid leukemia (CML) patients, as well as in healthy control subsets. Using a FISH technique, we estimated the random aneuploidy and telomere capture of the 21q22, SNRPN, and 15qter loci. Higher aneuploidy rates were found in the leukocytes of CLL and CML patients, compared with the control group, for the 21q22 and SNRPN loci. There was no difference in the aneuploidy rate between the CML and CLL groups. Telomere capture was found in the two groups (CLL and CML), but not in the control group. We propose that the telomere capture phenomenon is much more common than has been reported in the literature; however, its prognostic significance is yet to be established.

To determine whether low maternal serum concentrations of human chorionic gonadotropin (hCG) were associated with poor pregnancy outcome.Between 1999 and 2000, 20,880 women underwent triple test screening in our hospital. The levels of hCG were detected by fluorescent immunoassay. Low hCG levels (or = 0.20 MoM in our center) were considered to be a marker for increased risk for trisomy 18 or adverse pregnancy outcome. Each patient completed a questionnaire regarding fetal karyotype, complications of pregnancy and pregnancy outcome.Low maternal serum concentrations of hCG were detected in 119 pregnancies (0.57%). Of these, 19 (16%) were found to be missed abortions. The distribution of the remaining 100 cases was as follows: 72% had an isolated low hCG level, 24% had a low hCG level and a combination of hCG + alpha-fetoprotein0.80 MoM, and 4% had a low hCG level and a combination of hCG + unconjugated estriol0.80 MoM. No trisomy 18 or other chromosomal abnormalities were detected in our patient population. However, there were perinatal complications.Based on our screened population, a combination of multiple analytes seemed to be a better marker than an isolated finding of low maternal serum concentrations of hCG with regards to abnormal fetal karyotype, specifically trisomy 18, although it did determine a high-risk group in terms of complications of pregnancy.

In this study we describe two patients with congenital absence of alpha-fetoprotein (AFP). The pathological examination results, including an immunohistochemical stain, which define qualitatively the levels of AFP detected by the biochemical studies and the comparative genomic hybridization (CGH) are enclosed. A description of the suggested functions of AFP and the means of its production are set forth. An explanation is suggested for the lack of symptoms in a newborn with undetectable levels of AFP and the mechanism by which this condition might occur.

Multiple myeloma represents a malignant proliferation of plasma cells derived from a single clone. It is well known that alkylating agents are capable of inducing myelodysplastic syndromes (MDS) and acute myelocytic leukemias (AML). This risk of both diseases in patients with multiple myeloma has been estimated to be 10-20% after 10 years. We aimed to evaluate the time course and the type of genetic abnormalities in melphalan-treated patients in the chronic stage of the disease. We applied fluorescence in situ hybridization methods with probes to 5q31 and 7q31 to mononuclear peripheral blood leukocytes of 18 melphalan-treated patients and compared the results to those of 8 untreated myeloma patients. We found three patients (17%) with a 5q31 deletion in their peripheral white blood cells, but no 7q31 deletion. These findings suggest that 5q- occurs before the overt development of MDS/AML and raise important concerns regarding long-term treatment of myeloma patients with alkylating agents. Also, the performance of cytogenetic evaluation should be considered before autologous transplantation. The clinical and biological implications of these findings should be evaluated in larger clinical and laboratory studies.

Benign cystic teratoma, also known as dermoid cyst, is the most common of all ovarian neoplasms accounting for 10%–20% of all ovarian cysts. Most have a normal 46,XX karyotype. Less than 2% can undergo malignant transformation. By using the comparative genomic hybridization technique on nine benign ovarian cysts we were able to show two cases with chromosomal aberrations not seen before in dermoid cysts but known to be involved in malignant ovarian tumors. We speculate that these are the tumors carrying the potential for malignant transformation.

Xeroderma pigmentosum (XP) is a rare autosomal recessive syndrome. Laboratory investigations have failed to detect any consistent anomaly in cells from XP heterozygotic subjects, although examples of behavior intermediate between normal and XP cells have been reported. To estimate random aneuploidy we applied fluorescence in situ hybridization (FISH) with alpha-satellite probes for chromosomes 8 and 9 and replication pattern for TP53 (p53), ERBB2 (HER-2/neu), and MYCN (N-MYC) loci and for the imprinted SNRPN locus. A significantly higher rate of aneuploidy rate was observed in XP patients and carriers than in controls. The asynchrony pattern was significantly higher in XP carriers and patients with all three coding loci analyzed and significantly lower in XP patients and carriers with the imprinted locus SNRPN than in the control group. Molecular cytogenetic parameters such as random aneuploidy and replication pattern, which are known to reflect chromosomal instability, may be part of the tumorigenesis process. In XP patients and carriers, this genetic instability may represent a potential for developing malignancies.

Comparative genomic hybridization (CGH) was applied to samples taken from various sites of placentas originating from complicated pregnancies: 24 with intrauterine growth restriction (IUGR), one with multiple fetal malformation, one with toxemia, one with hydrocephalus and two with undetectable maternal serum alpha-fetoprotein (MSAFP). One of the most common aberrations in the IUGR cases was the addition of a whole or part of the X chromosome. Other aberrations such as additional Y chromosome or of 13(q22) or loss of chromosome 17 also appeared in different cases. In one IUGR case trisomy 8 (in one site) and 47,XXY (in all sites) were detected. In the two cases with undetectable MSAFP monosomy 16 was found. Some of the results were also confirmed by the FISH technique. In all the control cases (six normal and five with aneuploidy) CGH concurred with the known karyotype. Our results demonstrate the usefulness of the CGH technique in the genetic evaluation of fresh and paraffin embedded placentas in problematic pregnancies even when morphology is normal. However, it is very important to take multiple samples from different sites of the placenta.

Objective A study was conducted to evaluate the sensitivity of combining a second trimester triple test and targeted ultrasound in order to detect Down syndrome in women undergoing amniocentesis over 35 years of age. Methods Women over 35 years of age underwent a triple test and an ultrasound examination for chromosomal markers immediately prior to genetic amniocentesis. Results One thousand and six women were examined. Four hundred and thirty seven were triple test-positive and in 195 cases ultrasonographic abnormalities were observed. Thirteen had Down syndrome and eight had other chromosomal abnormalities. All women with Down syndrome babies were triple test-positive and seven also had ultrasonographic markers. Three of eight women who had babies with chromosomal aberrations other then Down syndrome were also triple test-positive. Conclusions The use of the triple test as a screening tool in our population would reduce the number of amniocenteses by 60%, while no cases of Down syndrome would be missed. Ultrasonographic markers have added little to this population. Three non-Down syndrome chromosomal abnormalities and two Down syndrome mosaic cases would be missed by this approach. Copyright © 2002 John Wiley & Sons, Ltd.

We describe the use of the fluorescence in situ hybridization (FISH) technique to detect residual Philadelphia chromosome-positive (Ph+) cells in a patient with blastic phase chronic myelogenous leukemia (CML) after aggressive cytoreductive treatment. The analysis was made in interphase nuclei because of the very small number of recognizable metaphases in leukemic patients. FISH was a reliable tool for the detection of chromosome translocations in interphase nuclei as compared with conventional cytogenetic and polymerase chain reaction (PCR) techniques.

Hepatocellular carcinoma (HCC) is a very common and highly malignant tumor, associated mainly with chronic viral hepatitis, cirrhosis of any cause, aflatoxin exposure and ethanol consumption. The aim of this study was to map genomic aberrations in HCC by a recently developed technique: comparative genomic hybridization (CGH). We applied CGH on 17 liver specimens, of which seven were HCCs. The rest were benign liver tumors, cirrhotic and normal livers, and other liver malignancies. Our study included mainly large tumors (mean size 10.5 cm) unrelated to viral hepatitis or cirrhosis. Our CGH analysis detected genomic imbalances in 42% of HCCs. The common aberrations included DNA gains of 1q, 9p, and 8q and DNA losses of 17p, 13q, 9q, 4q, and 11q. Also, we detected trisomies 8, 9, 18 and 21, which have not been reported previously. Gains and losses of DNA found in this study probably involve oncogenes and tumor suppressor genes that play a role in the puzzle of hepatocarcinogenesis. This study also suggests a possible link between the size of the tumor and the burden of genetic changes.

Chronic lymphocytic leukemia (CLL) is the most common leukemia in humans, with the major cytogenetic aberrations of trisomy 12 and deletion of 13q14. This study examined the influence of these aberrations on general gene replication. The study group included three subgroups: (1) 15 CLL patients, (2) 4 CLL patients with trisomy 12, (3) 3 CLL patients with deletions in 13q14. Five healthy individuals served as a control group. Monocolor fluorescence in situ hybridization (FISH) with probes for c-myc, HER-2/neu, and p53 was applied to lymphocyte nuclei for the evaluation of replication timing. Asynchronous replication (SD) rate was significantly higher in all CLL patients (P0.01) when compared to the control group and was even higher in the group of CLL patients with trisomy 12 and 13q14 deletion (P0.01). The asynchrony rate was significantly higher in cells with trisomy 12 for all three probes analyzed, compared to "healthy" cells in the same patients (P0.001). To conclude, in CLL patients with a chromosomal aberration such as trisomy 12 and 13q14 deletion we were able to demonstrate a high rate of asynchrony of replication. The high correlation between cells with trisomy 12 and SD pattern could reflect direct influence of the aberration on gene replication and cell cycle control.

Objective Essential mixed cryoglobulinemia (EMC) type II is associated with hepatitis C virus (HCV) in 90% of the patients with this disorder. A significant subset of these patients is at risk to develop non-Hodgkin lymphoma (NHL). The objective of this study was to examine whether the presence of EMC, a presumably premalignant step of lymphoproliferation, is associated with changes in the replication state of normal structural genes. Materials and Methods The study group included three subgroups: (1) seven patients with HCV without EMC; (2) eight patients with HCV associated with EMC. 3. Seven patients with follicular lymphoma; and (3) six healthy individuals served as control group. Monocolor fluorescent in situ hybridization (FISH) with probes to p53, RB-1, and 21q22 was applied to leukocytes nuclei for the evaluation of replication timing. Results Asynchronous replication (SD) rate was similar in patients with NHL and those with HCV associated with EMC and both are significantly higher when compared to patients with HCV without EMC and to normal controls (p 0.01) for each comparison. This held true for all studied loci (21q22, RB-1, and p53). Patients infected by HCV (but without EMC) had a significantly higher rate of asynchronous pattern in comparison with healthy controls (p 0.01). Conclusions Patients with a "premalignant" clinical condition HCV with EMC already demonstrate asynchronous type of replication which is similar to patients who already have an established malignant disease (i.e., NHL). In the future, replication may be used to assess the risk of malignant transformation in patients with "benign" proliferation.

The risk of myelodysplastic syndrome (MDS) or acute myeloblastic leukemia (AML) in patients with multiple myeloma has been estimated to be 10-20% after 10 years. Most myeloma patients develop MDS/AML after 3-4 years of treatment with alkylating agents, mainly melphalan; chromosomes 5 and 7 are most frequently involved. We studied 14 patients with myeloma by fluorescence in situ hybridization (FISH) with a probe to 5q31 (the critical area of deletion on chromosome 5) to verify whether deletion of 5q31 occurs during the course of stable, uncomplicated myeloma, and to assess the clinical importance of this abnormality. We found 2 patients (14%) with deletion of 5q31 in 30-40% of their peripheral white blood cells. One patient with this deletion received a high cumulative amount of melphalan, and the other patient was treated with multiple alkylating agents, including melphalan. In these patients, no clinical or laboratory evidence of transformation occurred 14 and 12 months after the finding of the aberration. These findings suggest that 5q-may occur months prior to the overt development of (t)-MDS/AML, and raise important concerns regarding the management of patients with this and similar aberrations, including modification of treatment and performance of cytogenetic evaluation prior to autologous or PSC transplantation. The clinical and biological implications of these findings should be evaluated in larger clinical and laboratory studies.

Nonrandom deletions of the long arm of chromosome 6 (6q) are associated with various lymphoid malignancies. It has been suggested that deletions of 6q25∼27, 6q21, and 6q23 typically occur in intermediate-grade, high-grade, and low-grade lymphomas, respectively. We used fluorescence in situ hybridization (FISH) to evaluate the occurrence of 6q27 deletion in chronic lymphatic leukemia (CLL) and multiple myeloma (MM). 6q27 deletion was detected in 21% of patients with CLL and in 28% of patients with MM. The percentage of cells containing deletions ranged between 25∼49. Two patients with MM had progressive disease and the aberration was detected in both. We conclude that FISH is a sensitive method to detect 6q27 deletion in lymphoproliferative disorders. Also, this deletion is not specific to intermediate-grade lymphomas, but occurs also in CLL and MM.

The study investigates the effect of sperm preincubation with FF on the fertilization rate in an IVF program. The oocytes inseminated with FF pretreated semen showed a significantly higher rate of fertilization. The percentage of cleavage and polypronuclear occurrence did not differ from the controls.

Intracranial hemorrhage during pregnancy is very rare. This catastrophic event has a poor prognosis and can contribute to maternal death for nonobstetrical reasons. The proper management during pregnancy remains controversial. Increasing awareness of the condition had been prompted by the fact that an acquired diagnosis is now usually available and a definitive and successful surgical procedure may be possible.

Objective Macrosomia at term is associated with increased maternal and neonatal morbidity, including a higher rate of cesarean delivery and shoulder dystocia. Induction of labor has been suggested as a means to prevent further weight gain and improve outcome. The aim of this study was to determine whether or not induction of labor in these cases improves maternal and neonatal outcome. Methods Patients at term with an ultrasonic fetal weight estimation of 4000–4500 g were prospectively randomized into two groups: induction of labor (group I) and expectant management (group II). Patients with diabetes, a previous cesarean delivery, or nonvertex presentation were excluded. Outcome variables included mode of delivery, arterial cord pH, presence of shoulder dystocia, brachial plexus injury, clavicular fracture, cephalohematoma, and intraventricular hemorrhage. Results Of 273 patients who were eligible for the study, 134 were randomized to group I and 139 to group II. Parity, gestational age, and fetal weight estimation were similar in the two groups. The neonates of group II patients were significantly heavier (4132.8 ± 347.4 versus 4062.8 ± 306.9 g; P = .024). The rate of cesarean delivery was 19.4% in group I and 21.6% in group II patients (not significant [NS]). Cord pH was similar in both groups. Shoulder dystocia was diagnosed in five group I and six group II patients (NS). None developed brachial plexus injury. There were two cases of mild, transient brachial plexus injury in group II patients without documented shoulder dystocia. Mild intraventricular hemorrhage was diagnosed in three of 44 group I and two of 31 group II neonates evaluated (NS). Conclusion In this prospective, randomized study, induction of labor for suspected macrosomia at term did not decrease the rate of cesarean delivery or reduce neonatal morbidity. Ultrasonic estimation of fetal weight between 4000 and 4500 g should not be considered an indication for induction of labor.

The karyotype of acute myeloid leukemia (AML) blasts has a major prognostic importance. However conventional cytogenetic studies of AML patients may fail to reveal chromosome abnormalities. Trisomy 8 is a common numerical abnormality in all AML subtypes. We evaluated the role of fluorescent in situ hybridization (FISH) in the detection of trisomy 8 in de-novo AML, and compared the results to chromosome analysis in some patients. Cytogenetic studies were performed in 9 of 12 patients. In three patients no metaphases were obtained. Of the remaining six, trisomy 8 was only detected in the metaphases of 1 patient. In contrast, 4 patients showed +8 with FISH and one had a borderline value. We conclude that FISH is a rapid and sensitive method to detect numerical aberrations in AML. In the future larger prospective studies should explore the biological and clinical application of the FISH method in different hematological malignancies.

608 Senescence in genotoxic amniocytes and stressed trophoblasts Tal Biron-Shental, Moshe Fejgin, Yehudit Sharon, Dvora Kidron, Aliza Amiel Meir Medical Center, Sackler School of Medicine, Tel Aviv University, Obstetrics and Gynecology, Kfar Saba, Israel, Meir Medical Center, Tel Aviv University, Genetic Institute, Kfar Saba, Israel, Bar Ilan University, Faculty of Life Science, Ramat Gan, Israel, Meir Medical Center, Sackler School of Medicine, Tel Aviv University, Pathology, Kfar Saba, Israel OBJECTIVE: Cellular senescence is a state of metabolic and cell cycle arrest evoked by exogenous or endogenous stressors, especially genotoxic stress. Some cases of senescence are characterized by formation of senescence-associated heterochromatin foci (SAHF). Short, dysfunctional telomeres lead to senescence. We have previously reported short telomeres in aneuploidy, intrauterine hypoxia (which causes intrauterine fetal growth restriction [IUGR]), and in uncontrolled gestational diabetes mellitus. We studied senescence in amniocytes and placentas from aneuploidic fetuses, in trophoblasts from IUGR fetuses, and from diabetic placentas, by assessing SAHF. STUDY DESIGN: Using DAPI staining, we counted the percentage of cells with increased nuclear fragmentation (SAHF). We assessed SAHF expression in trophoblasts from placental biopsies (n 7) and in amniocytes (n 10) from known trisomy-21 fetuses and compared the results to control placentas (n 6) and to amniocytes (n 10) from fetuses with a known normal karyotype. We also assessed the presence of SAHF in trophoblasts from pregnancies complicated by IUGR (n 12) and from pregnancies complicated by uncontrolled diabetes mellitus [DM] (n 10) and compared them to gestational-agematched controls from uncomplicated pregnancies (n 12). RESULTS: A significantly higher rate of cells with SAHF was found in trisomy-21 placentas and amniocytes, as well as in IUGR and in trophoblasts from pregnancies with uncontrolled diabetes compared to the control samples (Figure). CONCLUSION: Increased SAHF formation in amniocytes and in trophoblasts that were previously found to express shorter telomeres emphasizes the correlation between short telomeres and senescence. These findings may be linked to other genetic instability parameters previously described in trisomy-21 and help explain intrauterine programming in pregnancies with IUGR and uncontrolled diabetes mellitus. 609 Insulin growth factor 1 (IGF-1) therapy restored IUGR induced alteration in pancreatic tissues Khaled Omar, Tarek Alsaied, Ahmed Lababidi, Helen Jones, Swathi Balaji, Sundeeb Keswani, Timothy Crombleholme, Mounira Habli Cincinnati Childrens Hospital medical center, Center for molecular fetal therapy, Cincinnati, OH, Cincinnati Childrens Hospital medical center, Pediatrics, Cincinnati, OH, Colorado Childrens Hospital medical center, Pediatric surgery, Aurora, CO OBJECTIVE: We demonstrated that intraplacental gene therapy with adenoviral insulin growth factor (Ad-hIGF1) corrects impaired birthweight and glucose regulatory response in a mouse model of placental insufficiency. Evidence from diabetic patients and Murine IUGR offspring demonstrates decreased area and mass of islets and beta cells. We hypothesized that one of the mechanisms of AdhIGf1 induced attenuation in IUGR related adult onset diabetes is by restoring pancreatic mass, beta and alpha cell ratio. STUDY DESIGN: Laparotomy was performed on pregnant C57BL/6J mice at gestational day 18 and pups were divided into 3 groups (n 3). Control: Sham operated; IUGR: by ligation of a branch of mesenteric uterine artery; IGF1-Treated: intra placental injection of Ad hIGF1 after ligation. Pups were delivered on day 20, cross-fostered to CD1 mice. At 32 weeks, Pancreas of the off springs were weighed, and histologically processed and whole mounted. After every 20 section, a 5 um consecutive primary and reference sections were obtained for the entire pancreas. Immunohistochemistry staining was performed for Beta (anti-insulin) and Alpha (anti-glucagon) cells, and area was quantified (design-based stereology). Data were analysed using ANOVA. RESULTS: Male IUGR demonstrated 22% significant reduction in pancreatic mass (230.6 5.8 vs 296 5.7 vs 290 10 mg, p 0.01), 84 % reduction in Islets/Pancreas percentage area (2.5 1.1% vs 14.8 2.6% vs 16 3.7%, p 0.02), 80% in Beta cell/Alpha cell percentage area and 12% in Beta cell/ Islets percentage area (Table) as compared to SHAM, which was restored to normal with IGF1 treatment. Total area of pancreas, Islets, beta and alpha cells did not show any significant differences among 3 groups. CONCLUSION: Intra-placental gene transfer of Ad-hIGF1 restored IUGR-induced alterations in pancreatic mass, beta and alpha cell ratio. These changes may represent a potential mechanism of fetal reprogramming by which (AdhIGF1) may attenuate fetal predisposition to adult onset Diabetes. Algorithm for stepwise, contingent Down syndrome screening www.AJOG.org Academic Issues, Antepartum Fetal, Clinical Ob, Fetus, Genetics, Hypertension, Med-Surg-Diseases, Operative Ob, U/S Poster Session IV

Trisomies 8 and 9 are the most common numerical chromosome abnormalities in polycythemia vera (PCV). Their role in the pathogenesis of the disease is unclear, however, as is their diagnostic or prognostic value. We evaluated fluorescent in situ hybridization as compared to chromosome analysis for the detection of trisomies 8 or 9 in peripheral blood cells of PCV patients. We demonstrated that FISH is a more sensitive method for the detection of the abnormalities. A positive correlation between the duration of the disease and trisomy 9 was found. FISH is a sensitive, convenient, and rapid method for the diagnosis and follow-up of chromosome aberrations in patients with PCV The application of FISH to a larger cohort of patients may provide valuable information regarding the role of the chromosomal aberrations in the initiation and progression of this disease.

pregnancies complicated with intrauterine growth restriction Tal Biron-Shental, Yudith Sharon, Dvora Kidron, Moshe D. Fejgin, Aliza Amiel Meir Medical Center, Tel Aviv University, Obstetrics and Gynecology, Kfar Saba, Israel, Meir Medical Center, Genetic Institute, Kfar Saba, Israel, Meir Medical Center, Tel Aviv University, Pathology, Kfar Saba, Israel, Meir Medical Center, Faculty of Life Sciences, Bar Ilan University, Genetic Institute, Kfar Saba, Israel OBJECTIVE: Telomeres are nucleoprotein structures sited at the termini of the chromosomes that protect them from end to end fusion and degradation. Intrauterine Growth Restriction (IUGR) correlates with placental telomere shortening and with senescence. Senescence is associated with changes in the nuclear chromatin known as senescence-associated heterochromatin foci (SAHFs). Shortened and therefore unstable chromosomes can be stabilized by transfer of telomeres from other chromosomes. This process is termed telomere capture. Our aim was to assess mechanisms of telomere shortening in placentas from pregnancies complicated with IUGR. STUDY DESIGN: Placental biopsies derived from 13 pregnancies complicated with IUGR and from 12 gestational-age matched uncomplicated pregnancies. Clinical and histopathological characteristics were collected to ensure IUGR was secondary to placental insufficiency. We estimated the percentage of trophoblasts with SAHFs and the expression of the telomere capture mechanism using Dapi staining and FISH technique. RESULTS: Significantly increased expression of SAHFs (p 0.013) and significantly increased appearance of the telomere capture mechanism (p 0.023) were observed in placental biopsies from pregnancies complicated with IUGR compared to the control samples. (table). CONCLUSION: These results support and explain the observations of telomere shortening in IUGR placentas. We suggest that these findings may play a role in the pathophysiology of IUGR, perhaps by promoting senescence in trophoblasts of IUGR placentas.

and improving insulin sensitivity. We sought to assess whether the addition of metformin to insulin treated pregnant women would improve glucose control and lower insulin requirements. STUDY DESIGN: This retrospective study included pregnant women with diabetes requiring 100 units of insulin to achieve euglycemia. Unbalanced glucose levels mandating further treatment ( 100 daily units of insulin) were divided into 2 groups: Group A treated with increasing doses of insulin solely. Group B addition of metformin 850mg bid along with insulin. The groups were compared for parameters of glucose control (number of daily hyperglycemia levels, time needed to achieve optimal control, maternal satisfaction on a scale from 1 to 5 and perinatal outcome (birthweight, macrosomia, shoulder dystocia, neonatal hypoglycemia). RESULTS: 64 pregnant women meeting the inclusion crtieria were included. Metformin was added to the insulin treatment in 24 women (group B) while the other 40 were administered with increasing doses of insulin to achieve euglycemia (group A). The two groups were similar with regards to maternal age, body mass index, uterine scar and percentage of pre-gestational diabetes. Mean daily insulin requirements were significantly higher in group A (1352.3 vs 1053.6 units, p 0.05). Optimal glycemic control was achieved earlier with the combined treatment (8.31.2 days vs 10.11.2 days, p 0.05) and was better tolerated by the parturients (satisfaction score of 4.3 vs 3.6, p 0.05). Obstetrical performance was similar between the two groups with similar birthweights (3485212gr vs 3387138gr), NICU admission rates, neonatal hypoglycemia and macrosomia. CONCLUSION: The addition of metformin to pregnant women with diabetes requiring large amounts of insulin resulted in more rapid glucose control and higher maternal satisfaction and should be considered in these cases.

Data concerning oncogene activation in CLL are very limited. When studied by Southern blot, rearrangements of bcl -1, bcl -2, and bcl -3 have been only infrequently reported. We evaluated the role of fluorescence in situ hybridization (FISH) in the detection of gene rearrangements in two CLL patients. We used multiple DNA probes, including those of chromosome 12, immunoglobulin heavy and light chains, and the oncogenes bcl -1, bcl -2, and bcl -3. Additionally, routine cytogenetic study was performed. In one patient, trisomy 12 and bcl -2 translocation were demonstrated by both methods, while trisomy 12 and bcl -1 translocation were seen in the second patient, who had a normal karyotype. Larger studies should evaluate the role of FISH in the detection of oncogene involvement in CLL and compare it with other molecular methods.

Detection of minimal residual disease is one of the major goals in bone marrow transplantation. We used a fluorescence in-situ hybridization technique to detect residual Philadelphia-chromosome positive cells in chronic myelogenous leukemia (CML) patients after sex-mismatch BMT. We analyzed the level of detection using probes for the BCR/ABL fusion product by comparison with results obtained with probes for the Y and X sex chromosomes. Detection of sex-mismatch chromosomes was significantly higher than that of the BCR/ABL translocation. In contrast, a higher specificity of residual tumor cell detection by the BCR/ABL probe was demonstrated because most of the sex-mismatch cells detected by FISH had a normal karyotype. Tumor-specific markers probes are thus superior and more accurate than sex-mismatch probes for detection of MRD in CML patients after BMT.

To assess prospectively the efficiency and safety of two extended spectrum cephalosporins used as pre-operative prophylaxis in nonelective cesarean sections, and compare the results to those of a third group of patients that received cefamezine post cord clamping.Two hundred and forty one patients undergoing a nonscheduled cesarean section were assigned to receive either cefonicid or ceftriaxone prior to skin incision. These patients were followed prospectively for infectious and fetal complications. The outcome of these patients was also compared with another group of 194 patients that received cefamezine prophylaxis post cord clamping, and whose data were collected retrospectively. Chi-square analysis of variance were performed with P0.05 considered significant.There were no significant differences in the febrile complications among the two groups of patients that received pre-operative prophylaxis. However, these patients had significantly less wound infections (P = 0.008) and a significantly shorter hospital stay (P0.001) than the patients who received their prophylactic antibiotics post cord clamping.Extended-spectrum cephalosporins, when given pre-operatively, are both effective and safe, and may have an advantage over intra-operative first generation cephalosporins in the reduction of post cesarean section infectious morbidity.

Burkitt's lymphoma cells exhibit the reciprocal balanced chromosomal translocation t(8:14) in 75% of patients. Cytogenetic analysis is time-consuming, requires in vitro culture with mitogens and enables to analyse a relatively small number of cells. We evaluated the role of multicolor fluorescent in situ hybridization (FISH) in the rapid detection of t(8:14) in Burkitt's lymphoma cells. FISH detected the JH/myc translocation in 100% of the cells of five cell lines carrying the classical t(8:14) and in fresh cells obtained from a newly diagnosed Burkitt's lymphoma patient. In contrast, the translocation was not detected in the Bajb cell line that does not carry t(8:14). We conclude that FISH is a rapid and reliable diagnostic tool for the detection of the JH/myc translocation in Burkitt's lymphoma patients.

A case report is presented in which a tubal pregnancy established after ET of frozen-thawed embryos in an amenorrheic woman had progressed for 3 weeks without ovarian steroid supplementation

Fifty-two women, who had 62 ovum aspiration cycles, received a progesterone (P) supplementation of 100 mg/day that was initiated 10 hours before human chorionic gonadotropin (hCG) administration and was continued over the following 6 days. Forty-eight women who had 74 ovum pick-ups, but did not get P, served as controls. Forty-four (84.6%) women of the treatment group, and 40 (83.3%) of the controls had ovum fertilization and embryo replacement. The fertilization and cleavage rates and the mean number of replaced embryos per embryo transfer (ET) cycle did not differ between the groups. Endometrial biopsies, from treatment group women with no fertilized eggs, which were taken 48 hours after ovum pick-up, mostly revealed an "advanced endometrial dating," in relation to the "day of hCG." Pregnancy rate per ET cycle for the treatment group was significantly higher than that of the controls; 41.2% versus 23.3%, respectively. It is concluded that the higher pregnancy rate resulted from an improvement in uterine receptivity.

Laparoscopic surgery is currently the preferred diagnostic and treatment modality for ectopic pregnancy (EP). Cornual pregnancy, a rare and dangerous form of EP, is usually treated by cornual resection or hysterectomy. We report the first case to be treated by laparoscopic surgery. A 27 year old, gravida 3 para 2, was diagnosed with a nine week right cornual pregnancy. Laparoscopy revealed a 3 x 4 cm bulge into the right cornual area. Both fallopian tubes were otherwise normal. The uterus was fixed by a grasping forceps and the right tube and ovarian ligament were coagulated to reduce the blood supply. The gestational area was entered via a 1 cm incision using unipolar scissors and electrocoagulation. The pregnancy contents were suctioned; cavity margins and placental bed were coagulated until hemostasis was secured. The uterus contracted. The posterior leaf of the broad ligament was flapped over the incision site and attached by coagulation points. Postoperative course was uneventful. A hysterosalpingogram performed 4 months post surgery revealed a normal uterine cavity and blocked right tube.

To the Editor.— Amniotic fluid embolism is a rare but deadly pathological entity that accounts for roughly 10% of all maternal deaths. 1,2 We describe a fatal outcome following induction of labor with prostaglandin. Report of a Case.— A 37-year-old white woman, gravida 4, para 2, aborta 1, was admitted to our labor ward at 42 weeks' gestation for induction of labor. Her current pregnancy was uncomplicated. Results of a genetic amniocentesis at 16 weeks were normal. At 42 weeks, the patient was admitted for an oxytocin challenge test. All vital signs were normal and the cervix was posterior and uneffaced. The results of her oxytocin challenge test were negative and she was discharged after her contractions stopped. The following day, she returned complaining of reduction in fetal movements. The nonstress test was reactive and results of physical examination were unchanged. In an attempt to "ripen" the cervix, 3 mg