Maternal obesity heritably perturbs offspring metabolism for three generations without serial programming.

Maternal obesity can program offspring metabolism across multiple generations. It is not known whether multigenerational effects reflect true inheritance of the induced phenotype, or are due to serial propagation of the phenotype through repeated exposure to a compromised gestational milieu. Here we sought to distinguish these possibilities, using the Avy mouse model of maternal obesity. In this model, F1 sons of obese dams display a predisposition to hepatic insulin resistance, which remains latent unless the offspring are challenged with a Western diet. We find that F2 grandsons and F3 great grandsons of obese dams also carry the latent predisposition to metabolic dysfunction, but remain metabolically normal on a healthy diet. Given that the breeding animals giving rise to F2 and F3 were maintained on a healthy diet, the latency of the phenotype permits exclusion of serial programming; we also confirmed that F1 females remained metabolically healthy during pregnancy. Molecular analyses of male descendants identified upregulation of hepatic Apoa4 as a consistent signature of the latent phenotype across all generations. Our results exclude serial programming as a factor in transmission of the metabolic phenotype induced by ancestral maternal obesity, and indicate inheritance through the germline, probably via some form of epigenetic inheritance.

Human APC2 localization and allelic imbalance.

A second adenomatous polyposis coli (APC)-like gene, APC2/APCL, was recently described and localized to chromosome 19. We have fine mapped APC2 to a small region of chromosome 19p13.3 containing markers D19S883 and WI-19632, a region commonly lost in a variety of cancers, particularly ovarian cancer. Interphase fluorescence in situ hybridization analysis revealed an APC2 allelic imbalance in 19 of 20 ovarian cancers screened and indicates that APC2 could be a potential tumor suppressor gene in ovarian cancer. When overexpressed in SKOV3 ovarian cancer cells, which express low levels of APC2, exogenous APC2 localized to the Golgi apparatus, actin-containing structures, and occasionally to microtubules. Antibodies against the NH2 terminus of human APC2 show that endogenous APC2 is diffusely distributed in the cytoplasm and colocalizes with both the Golgi apparatus and actin filaments. APC2 remained associated with actin filaments after treatment with the actin-disrupting agent, cytochalasin D. These results suggest that APC2 is involved in actin-associated events and could influence cell motility or adhesion through interaction with actin filaments, as well as functioning independently or in cooperation with APC to down-regulate beta-catenin signaling.

Amniotic fluid androgens and estrogens in midgestation.

Amniotic fluid androgen and estrogen levels associated with 48 male and 72 female fetuses between 14 and 20 weeks of gestation were measured. Amniotic fluid testosterone levels were significantly higher (P less than 0.001) in the male (224 +/- 11 pg/ml) than the female fetuses (39 +/- 2 pg/ml) with no overlap of values. Amniotic fluid androstenedione concentrations were also significantly higher (P less than 0.001) with male (1024 +/- 53 pg/ml) than female fetuses (668 +/- 39 pg/ml), but there was overlap. There was no difference between anmiotic fluid dehydroepiandrosterone levels for the two sexes. Estrone concentrations were slightly but not significantly higher with the male (353 +/- 33 pg/ml) than with female fetuses (331 +/- 28 pg/ml), while estradiol concentrations were significantly higher (P =0.002) with the female (96 +/- 8 pg/ml) than male (64 +/- 4 pg/ml) fetuses. It is interpreted that the higher amniotic fluid testosterone and androstenedione levels for the male fetuses reflect fetal testicular secretion. The significantly higher estradiol concentrations for the female fetuses may reflect early ovarian secretion.

NRG-3 in human breast cancers: activation of multiple erbB family proteins.

Ligands of the EGF/Heregulin family control the growth of epithelial cells by binding to receptors of the erbB family. By searching a large database of cDNA sequences at Human Genome Sciences Inc. we have identified a new encoded protein sequence containing all the conserved elements of the EGF/Heregulin family. The same sequence has recently been independently identified as NRG-3. The EGF-like domain of NRG-3 was generated as a recombinant protein in E. coli and used to test the specificity of receptor binding. In human breast cancer cells and in 32D cells transfected by erbB family members, NRG-3 activated multiple erbB family members. These include EGF receptor (erbB1) and erbB4 when expressed individually and erbB2 and erbB3 when expressed together. Recombinant NRG-3 altered the growth of human breast cancer cells growing in vitro. NRG-3 was expressed in cell lines derived from breast cancer. These results indicate that NRG-3 is a potential regulator of normal and malignant breast epithelial cells in vivo.

Ligneous cellulitis associated with an IUD.

Many reports have documented serious pelvic infection associated with an IUD. A case of ligneous pelvic cellulitis with cutaneous sinus tract formation, hydronephrosis, and ureteral obstruction in a patient with an IUD is presented. This is an uncommon manifestation of chronic pelvic infection and responded to antibiotics and steroid administration. The origin and treatment of ligneous cellulitis are discussed.

Amniotic fluid testosterone levels in midpregnancy.

Amniotic fluid testosterone levels were measured on specimens obtained between 12 and 25 weeks' gestation from 58 male-fetus and 77 female-fetus pregnancies. For the male fetuses the mean +/- SE amniotic fluid testosterone level of 223 +/- 10 pg/ml was significantly higher (P less than 0.001) than the concentration found for the female fetuses (40 +/- 2 pg/ml). The ranges were 104-424 and 18-82 pg/ml, respectively, for the same fetuses. In the male fetuses the highest mean level was found during the 17th gestational week, but the mean level observed during any 1 week was not significantly differenf from any other week for both sexes. These data are consistent with the concept that amniotic fluid testosterone levels may be a rapid and effective method for establishing fetal sex in utero during midgestation.

Diagnosis of a triploid fetus at genetic amniocentesis.

Nearly 20% of spontaneously aborted fetuses with chromosomal errors have been shown to have a triploid karyotype. Such a fetus is rarely encountered during the course of an amniocentesis at midtrimester. Because amniotic fluid cell polyploidy is an occasional finding among normal fetuses at midtrimester, the following report illustrates the importance of gathering corroborative evidence of an affected fetus before arriving at a prenatal diagnosis.

K-shell spectroscopy of an independently diagnosed uniaxially expanding laser-produced aluminum plasma.

We present detailed spectroscopic analysis of the primary K-shell emission lines from a uniaxially expanding laser-produced hydrogenic and heliumlike aluminum plasma. The spectroscopic measurements are found to be consistent with time-dependent hydrodynamic properties of the plasma, measured using Thomson scattering and shadowgraphy. The K-shell population kinetics code FLY with the measured hydrodynamic parameters is used to generate spectra that are compared to the experimental spectra. Excellent agreement is found between the measured and calculated spectra for a variety of experimental target widths employed to produce plasmas with different optical depths. The peak emission from the hydrogenic Lyman series is determined to be from a temporal and spatial region where the hydrodynamic parameters are essentially constant. This allows a single steady-state solution of FLY to be used to deduce the electron temperature and density, from the measured line ratios and linewidths, for comparison with the Thomson and shadowgraphy data. These measurements are found to agree well with time-dependent calculations, and provide further validation for the FLY calculations of the ionization and excitation balance for a K-shell aluminum plasma. We also discuss the possible application of this data as a benchmark for hydrodynamic simulations and ionization/excitation balance calculations.

The utility of flexible sigmoidoscopy after a computerized tomographic colonography revealing only rectosigmoid lesions.

BACKGROUND: Identifying polyps by computerized tomographic colonography typically prompts colonoscopy, increasing its cost, risk and inconvenience. Many polyps are confined to the rectosigmoid and theoretically amenable to resection via flexible sigmoidoscopy. AIM: To determine the prevalence of advanced proximal colonic neoplasia when computerized tomographic colonography reveals only rectosigmoid polyps, and characterize the yield of polypectomy via flexible sigmoidoscopy in such patients. METHODS: Subjects underwent computerized tomographic colonography and colonoscopy with segmental unblinding. Patients with only rectosigmoid findings by computerized tomographic colonography were identified retrospectively. Flexible sigmoidoscopy findings were estimated by including lesions distal to the descending/sigmoid colon junction during colonoscopy. Proximal lesions were also reviewed. Advanced lesions were defined as: adenocarcinoma, tubular adenoma >1 cm, > or =3 tubular adenomas, tubulovillous histology or high-grade dysplasia. RESULTS: By computerized tomographic colonography, 15% (203 of 1372) had only rectosigmoid polyps. Concomitant lesions in the proximal colon were seen in 32% (64 of 203) during colonoscopy. Advanced proximal neoplasia occurred in 2% (three of 203) with only rectosigmoid polyps on computerized tomographic colonography. CONCLUSIONS: Using flexible sigmoidoscopy to follow-up computerized tomographic colonography demonstrating only rectosigmoid polyps would eliminate 15% of subsequent colonoscopies. This strategy carries a small risk of missed proximal advanced neoplasia. This miss rate appears comparable to that of colonoscopy alone. Further study on the cost-effectiveness of this approach is warranted.

Long duration backlighter experiments at Omega.

We have successfully demonstrated a 7.5 ns duration pinhole-apertured backlighter at the Omega laser facility. Pinhole-apertured point-projection backlighting for 8 ns will be useful for imaging evolving features in experiments at the National Ignition Facility. The backlighter consisted of a 20 microm diameter pinhole in a 75 microm thick Ta substrate separated from a Zn emitter (9 keV) by a 400 microm thick high-density carbon piece. The carbon prevented the shock from the laser-driven surface from reaching the substrate before 8 ns and helped minimize x-ray ablation of the pinhole substrate. Grid wires in x-ray framing camera images of a gold grid have a source-limited resolution significantly smaller than the pinhole diameter due to the high aspect ratio of the pinhole, but do not become much smaller at late times.

Demonstration of the density dependence of x-ray flux in a laser-driven hohlraum.

Experiments have been conducted using laser-driven cylindrical hohlraums whose walls are machined from Ta2O5 foams of 100 mg/cc and 4 g/cc densities. Measurements of the radiation temperature demonstrate that the lower density walls produce higher radiation temperatures than the high density walls. This is the first experimental demonstration of the prediction that this would occur [M. D. Rosen and J. H. Hammer, Phys. Rev. E 72, 056403 (2005)10.1103/PhysRevE.72.056403]. For high density walls, the radiation front propagates subsonically, and part of the absorbed energy is wasted by the flow kinetic energy. For the lower wall density, the front velocity is supersonic and can devote almost all of the absorbed energy to heating the wall.

Abortion and menstrual extraction for the ambulatory patient.

PIP: Complications attending menstrual regulation, office suction and curettage abortion, and ambulatory hospital suction and curettage abortion are discussed and minimum criteria for outpatient abortion are sought. Guidelines established by the American College of Obstetricians and Gynecologists for the performance of abortion are reiterated. Uterine perforation, pelvic infections, hemorrhage, and incomplete evacuation are complications which should not exceed usual hospital rates when abortion is performed in an office. The possibilities that dilatation of the cervix in abortion may result in an increased risk of future stillbirth deliveries and premature delivery, that secondary infertility can result from subclinical salpingitis or the development of intrauterine synechia, and that an increased risk of ectopic pregnancy may result from abortion are long-term complications that must be considered. Standards, preoperative procedures, equipment and operative technique, and postoperative procedures and considerations in menstrual regulation are presented. The usual procedures for office and hospital suction curettage abortion are presented and discussed.^ieng

Development of Leishmania chagasi (Kinetoplastida: Trypanosomatidae) in the second blood-meal of its vector Lutzomyia longipalpis (Diptera: Psychodidae).

Light and electron microscopy investigations were carried out to compare the development of Leishmania chagasi in Lutzomyia longipalpis females that took a second blood-meal and others that took a single blood-meal. The establishment of the parasite in the foregut and the thoracic midgut of the vector was not severely affected by the intake of the second blood-meal. At 3 days after the intake of the second blood-meal, a rapid increase in the proportion of metacyclic promastigotes detected in the midgut and proboscis of the vector was noticed. No similar increase in the proportion of metacyclic promastigotes observed in the females that took a single blood-meal was noticeable. The results indicate a higher probability of transmission of the parasite to the vertebrate host at the third bite of the vector than at the second bite.

Dynamic changes in the distribution of cytoplasmic myosin during Drosophila embryogenesis.

Dramatic changes in the localization of conventional non-muscle myosin characterize early embryogenesis in Drosophila melanogaster. During cellularization, myosin is concentrated around the furrow canals that form the leading margin of the plasma membrane as it plunges inward to package each somatic nucleus into a columnar epithelial cell. During gastrulation, there is specific anti-myosin staining at the apical ends of those cells that change shape in regions of invagination. Both of these localizations appear to result from a redistribution of a cortical store of maternal myosin. In the preblastoderm embryo, myosin is localized to the egg cortex, sub-cortical arrays of inclusions, and, diffusely, the yolk-free periplasm. At the syncytial blastoderm stage, myosin is found within cytoskeletal caps associated with the somatic nuclei at the embryonic surface. Following the final syncytial division, these myosin caps give rise to the myosin rings observed during cellularization. These distributions are observed with both whole immune serum and affinity-purified antibodies directed against Drosophila non-muscle myosin heavy chain. They are not detected in embryos stained with anti-Drosophila muscle myosin antiserum or with preimmune serum. Although immunolocalization can only suggest possible function, these myosin localizations and the coincident changes in cell morphology are consistent with a key role for non-muscle myosin in powering cellularization and gastrulation during embryogenesis.

The sialomucin CD34 is expressed on hematopoietic cells and blood vessels during murine development.

The processes of angiogenesis and hematopoiesis require a high degree of coordination during embryogenesis. Whereas much is understood about the development of the vascular system in avian embryos, little information has been attained in mammals, predominantly because there are no specific markers for either blood vessels or hematopoietic cells in any developing mammalian system. We have recently shown that murine CD34 (mCD34) is expressed on the vascular endothelium in all organs and tissues of the adult mouse as well as on a small percentage of presumably hematopoietic stem cells in the bone marrow and fetal liver. Here we show that mCD34 is also expressed on the endothelium of blood vessels and on a subset of hematopoietic-like cells throughout murine development. mCD34 is first observed on the yolk sac endothelium of day 7.5 embryos and on a subset of hematopoietic cells within these yolk sacs. mCD34 expression is maintained on vessels and hematopoietic cells in all organs and tissues throughout embryogenesis. In addition, mCD34 is localized on growth conelike filopodial processes that appear at the budding edge of newly sprouted capillaries. Double staining of capillaries for mCD34 and laminin shows that these growth conelike processes seem to be free of laminin, whereas the formed capillaries seem to be coated with this extracellular matrix protein. Analysis of vessels in developing brain shows that these filopodial processes seem to be directed toward the ventricular epithelium, a previously described site of vascular endothelial growth factor synthesis. Finally, we show that the vascular structures of developing murine embryoid bodies also express mCD34. These data suggest that mCD34 is a useful marker for the analysis of the development of the blood vascular system in murine embryos.

Amniocentesis for antenatal diagnosis. Review of problems and outcomes in a large series.

The results of amniocentesis for diagnostic genetic studies on 242 patients are presented in detail. These suggest that obtaining fluid free from contaminating red blood cells is important to successful cell culture. The data also suggest that it is technically more difficult to obtain such fluid tham might be expected, despite the use of ultrasonic localization and experienced operators. Nevertheless, 97 per cent of patients who elect to complete antenatal diagnosis will ultimately obtain the information sought, in time to intervene medically if desired. Moreover, the act of obtaining this information is shown to be without significant risk to the ultimate outcome of pregnancy.

Fetal exsanguination and other vascular injuries from midtrimester genetic amniocentesis.

A case of fetal exsanguination after midtrimester genetic amniocentesis is reported. Examination of the ration of fetal-to-meternal red cells in the amniotic fluid obtained by amniocentesis for genetic study from 242 consecutive patients, including the reported case, suggests that fetal hemorrhage is relatively common and difficult to avoid during this procedure. It also appears that hemorrhage occurs significantly more oftern when the placenta is located anteriorly. Reasons as to why the accident is rarely fatal are considered, and some recommendations for patient counselling and management are discussed.

Results of microsurgical treatment of tubal infertility and early second-look laparoscopy in the post-pelvic inflammatory disease patient: implications for in vitro fertilization.

Sixty-nine patients with tubal infertility secondary to pelvic inflammatory disease were surgically treated by one of three infertility surgeons, who used microsurgery for repair of the tubal pathologic condition and early second-look laparoscopy 6 to 30 days postoperatively for lysis of postoperative adhesions. No patient was included in this group whose disease was thought to have originated from endometriosis or prior abdominal surgery. The average follow-up time was 43.1 months (range 12 to 85.9). Nine patients were excluded from the analysis. Pregnancy outcome by procedure, expressed as the percentage of patients conceiving, was as follows: adhesiolysis, 69% (61% term, 8% ectopic); fimbrioplasty, 35% (25% term, 10% ectopic); salpingostomy, 30% (18% term, 12% ectopic); and cornual implantation, 60% (40% term, 20% ectopic). No added therapeutic value could be attributed to the use of early second-look laparoscopy. Given the relatively poor outcome of fimbrioplasty and salpingostomy, it may be prudent to advise patients with bilateral partial and/or total tubal occlusion against tuboplasty in favor of in vitro fertilization and embryo transfer.