Search

Free-electron lasers (FEL) hold the potential to revolutionize structural biology by producing X-ray pules short enough to outrun radiation damage, thus allowing imaging of biological samples without the limitation from radiation damage. Thus, a major part of the scientific case for the first FELs was three-dimensional (3D) reconstruction of non-crystalline biological objects. In a recent publication we demonstrated the first 3D reconstruction of a biological object from an X-ray FEL using this technique. The sample was the giant Mimivirus, which is one of the largest known viruses with a diameter of 450 nm. Here we present the dataset used for this successful reconstruction. Data-analysis methods for single-particle imaging at FELs are undergoing heavy development but data collection relies on very limited time available through a highly competitive proposal process. This dataset provides experimental data to the entire community and could boost algorithm development and provide a benchmark dataset for new algorithms.

We present a proof-of-concept three-dimensional reconstruction of the giant mimivirus particle from experimentally measured diffraction patterns from an x-ray free-electron laser. Three-dimensional imaging requires the assembly of many two-dimensional patterns into an internally consistent Fourier volume. Since each particle is randomly oriented when exposed to the x-ray pulse, relative orientations have to be retrieved from the diffraction data alone. We achieve this with a modified version of the expand, maximize and compress algorithm and validate our result using new methods.

Ten years ago, the discovery of Mimivirus, a virus infecting Acanthamoeba, initiated a reappraisal of the upper limits of the viral world, both in terms of particle size (>0.7 micrometers) and genome complexity (>1000 genes), dimensions typical of parasitic bacteria. The diversity of these giant viruses (the Megaviridae) was assessed by sampling a variety of aquatic environments and their associated sediments worldwide. We report the isolation of two giant viruses, one off the coast of central Chile, the other from a freshwater pond near Melbourne (Australia), without morphological or genomic resemblance to any previously defined virus families. Their micrometer-sized ovoid particles contain DNA genomes of at least 2.5 and 1.9 megabases, respectively. These viruses are the first members of the proposed "Pandoravirus" genus, a term reflecting their lack of similarity with previously described microorganisms and the surprises expected from their future study.

Mimivirus is one the largest DNA virus identified so far, infecting several Acanthamoeba species. Analysis of its genome revealed the presence of a nine-gene cluster containing genes potentially involved in glycan formation. All of these genes are co-expressed at late stages of infection, suggesting their role in the formation of the long fibers covering the viral surface. Among them, we identified the L136 gene as a pyridoxal phosphate-dependent sugar aminotransferase. This enzyme was shown to catalyze the formation of UDP-4-amino-4,6-dideoxy-D-glucose (UDP-viosamine) from UDP-4-keto-6-deoxy-D-glucose, a key compound involved also in the biosynthesis of L-rhamnose. This finding further supports the hypothesis that Mimivirus encodes a glycosylation system that is completely independent of the amoebal host. Viosamine, together with rhamnose, (N-acetyl)glucosamine, and glucose, was found as a major component of the viral glycans. Most of the sugars were associated with the fibers, confirming a capsular-like nature of the viral surface. Phylogenetic analysis clearly indicated that L136 was not a recent acquisition from bacteria through horizontal gene transfer, but it was acquired very early during evolution. Implications for the origin of the glycosylation machinery in giant DNA virus are also discussed.

Mimivirus, a DNA virus infecting acanthamoeba, was for a long time the largest known virus both in terms of particle size and gene content. Its genome encodes 979 proteins, including the first four aminoacyl tRNA synthetases (ArgRS, CysRS, MetRS, and TyrRS) ever found outside of cellular organisms. The discovery that Mimivirus encoded trademark cellular functions prompted a wealth of theoretical studies revisiting the concept of virus and associated large DNA viruses with the emergence of early eukaryotes. However, the evolutionary significance of these unique features remained impossible to assess in absence of a Mimivirus relative exhibiting a suitable evolutionary divergence. Here, we present Megavirus chilensis, a giant virus isolated off the coast of Chile, but capable of replicating in fresh water acanthamoeba. Its 1,259,197-bp genome is the largest viral genome fully sequenced so far. It encodes 1,120 putative proteins, of which 258 (23%) have no Mimivirus homologs. The 594 Megavirus/Mimivirus orthologs share an average of 50% of identical residues. Despite this divergence, Megavirus retained all of the genomic features characteristic of Mimivirus, including its cellular-like genes. Moreover, Megavirus exhibits three additional aminoacyl-tRNA synthetase genes (IleRS, TrpRS, and AsnRS) adding strong support to the previous suggestion that the Mimivirus/Megavirus lineage evolved from an ancestral cellular genome by reductive evolution. The main differences in gene content between Mimivirus and Megavirus genomes are due to (i) lineages specific gains or losses of genes, (ii) lineage specific gene family expansion or deletion, and (iii) the insertion/migration of mobile elements (intron, intein).

Single-particle experiments using X-ray Free Electron Lasers produce more than 10(5) snapshots per hour, consisting of an admixture of blank shots (no particle intercepted), and exposures of one or more particles. Experimental data sets also often contain unintentional contamination with different species. We present an unsupervised method able to sort experimental snapshots without recourse to templates, specific noise models, or user-directed learning. The results show 90% agreement with manual classification., (© 2011 Optical Society of America)

X-ray lasers offer new capabilities in understanding the structure of biological systems, complex materials and matter under extreme conditions. Very short and extremely bright, coherent X-ray pulses can be used to outrun key damage processes and obtain a single diffraction pattern from a large macromolecule, a virus or a cell before the sample explodes and turns into plasma. The continuous diffraction pattern of non-crystalline objects permits oversampling and direct phase retrieval. Here we show that high-quality diffraction data can be obtained with a single X-ray pulse from a non-crystalline biological sample, a single mimivirus particle, which was injected into the pulsed beam of a hard-X-ray free-electron laser, the Linac Coherent Light Source. Calculations indicate that the energy deposited into the virus by the pulse heated the particle to over 100,000 K after the pulse had left the sample. The reconstructed exit wavefront (image) yielded 32-nm full-period resolution in a single exposure and showed no measurable damage. The reconstruction indicates inhomogeneous arrangement of dense material inside the virion. We expect that significantly higher resolutions will be achieved in such experiments with shorter and brighter photon pulses focused to a smaller area. The resolution in such experiments can be further extended for samples available in multiple identical copies.

Mimivirus, a virus infecting Acanthamoeba, is the prototype of the Mimiviridae, the latest addition to the nucleocytoplasmic large DNA viruses. The Mimivirus genome encodes close to 1000 proteins, many of them never before encountered in a virus, such as four amino-acyl tRNA synthetases. To explore the physiology of this exceptional virus and identify the genes involved in the building of its characteristic intracytoplasmic "virion factory," we coupled electron microscopy observations with the massively parallel pyrosequencing of the polyadenylated RNA fractions of Acanthamoeba castellanii cells at various time post-infection. We generated 633,346 reads, of which 322,904 correspond to Mimivirus transcripts. This first application of deep mRNA sequencing (454 Life Sciences [Roche] FLX) to a large DNA virus allowed the precise delineation of the 5' and 3' extremities of Mimivirus mRNAs and revealed 75 new transcripts including several noncoding RNAs. Mimivirus genes are expressed across a wide dynamic range, in a finely regulated manner broadly described by three main temporal classes: early, intermediate, and late. This RNA-seq study confirmed the AAAATTGA sequence as an early promoter element, as well as the presence of palindromes at most of the polyadenylation sites. It also revealed a new promoter element correlating with late gene expression, which is also prominent in Sputnik, the recently described Mimivirus "virophage." These results-validated genome-wide by the hybridization of total RNA extracted from infected Acanthamoeba cells on a tiling array (Agilent)--will constitute the foundation on which to build subsequent functional studies of the Mimivirus/Acanthamoeba system.

Millions of American girls and women have been drawn to smoking by an industry that has been clearly and systematically targeting women of all ages and life circumstances. Tobacco marketing strategies skillfully link cigarette use to typical female values. Biologically speaking, women are especially vulnerable to the legion of health problems of tobacco use. Smoking is a critical hazard for women in their reproductive years, particularly when they are pregnant.

Background: Aspergillus fumigatus, a saprophytic mould, is responsible for life-threatening, invasive pulmonary diseases in immunocompromised hosts. The role of the airway epithelium involves a complex interaction with the inhaled pathogen. Antimicrobial peptides with direct antifungal and chemotactic activities may boost antifungal immune response., Results: The inducible expression of defensins by human bronchial epithelial 16HBE cells and A549 pneumocyte cells exposed to A. fumigatus was investigated. Using RT-PCR and real time PCR, we showed an activation of hBD2 and hBD9 defensin genes: the expression was higher in cells exposed to swollen conidia (SC), compared to resting conidia (RC) or hyphal fragments (HF). The kinetics of defensin expression was different for each one, evoking a putative distinct function for each investigated defensin. The decrease of defensin expression in the presence of heat-inactivated serum indicated a possible link between defensins and the proteins of the host complement system. The presence of defensin peptide hBD2 was revealed using immunofluorescence that showed a punctual cytoplasmic and perinuclear staining. Quantification of the cells stained with anti hBD2 antibody demonstrated that SC induced a greater number of cells that synthesized hBD2, compared to RC or HF. Labelling of the cells with anti-hBD-2 antibody showed a positive immunofluorescence signal around RC or SC in contrast to HF. This suggests co-localisation of hBD2 and digested conidia. The HBD2 level was highest in the supernatants of cells exposed to SC, as was determined by sandwich ELISA. Experiments using neutralising anti-interleukine-1beta antibody reflect the autocrine mechanism of defensin expression induced by SC. Investigation of defensin expression at transcriptional and post-transcriptional levels demonstrated the requirement of transcription as well as new protein synthesis during A. fumigatus defensin induction. Finally, induced defensin expression in primary culture of human respiratory cells exposed to A. fumigatus points to the biological significance of described phenomena., Conclusion: Our findings provide evidence that respiratory epithelium might play an important role in the immune response during Aspergillus infection. Understanding the mechanisms of regulation of defensin expression may thus lead to new approaches that could enhance expression of antimicrobial peptides for potential therapeutic use during aspergillosis treatment.

Particulate atmospheric pollutants interact with the human airway epithelium, which releases cytokines, chemokines, and EGF receptor (EGFR) ligands leading to proinflammatory responses. There is little information concerning the short-term effects of EGFR activation by extracellular ligands on ionic regulation of airway surface lining fluids. We identified in the membrane of human epithelial bronchial cells (16HBE14o(-) line) an endogenous calcium- and voltage-dependent, outwardly rectifying small-conductance chloride channel (CACC), and we examined the effects of EGF on CACC activity. Ion channel currents were recorded with the patch-clamp technique. In cell-attached membrane patches, CACC were activated by exposure of the external surface of the cells to physiological concentrations of EGF without any change in cytosolic Ca(2+) concentration ([Ca(2+)](i)) and inhibited by tyrphostin AG-1478 (an inhibitor of EGFR that also blocks EGF-dependent Src family kinase activation). EGF activation of c-Src protein in 16HBE14o(-) cells was observed, and the signaling pathway elicited by EGFR was blocked by tyrphostin AG-1478. In excised inside-out membrane patches CACC were activated by exposure of the cytoplasmic face of the channels to the human recombinant Src(p60(c-src)) kinase with endogenous or exogenous ATP and inhibited by lambda-protein phosphatase. Secretion of EGFR ligands by epithelial airway cells exposed to pollutants would then elicit a rapid and direct ionic response of CACC mediated by EGFR activation via a Src kinase family-dependent signaling pathway.

Cystathionine beta synthase deficiency induces hyperhomocysteinemia which is considered as a risk factor for vascular diseases. Studies underlined the importance of altered cellular redox reactions in hyperhomocysteinemia-induced vascular pathologies. Nevertheless, hyperhomocysteinemia also induces hepatic dysfunction which may accelerate the development of vascular pathologies by modifying cholesterol homeostasis. The aim of the present study was to analyze the modifications of redox state in the liver of heterozygous cystathionine beta synthase-deficient mice, a murine model of hyperhomocysteinemia. In this purpose, we quantified levels of reactive oxygen and nitrogen species and we assayed activities of main antioxidant enzymes. We found that cystathionine beta synthase deficiency induced NADPH oxidase activation. However, there was no accumulation of reactive oxygen (superoxide anion, hydrogen peroxide) and nitrogen (nitrite, peroxynitrite) species. On the contrary, hepatic hydrogen peroxide level was decreased independently of an activation of glutathione-dependent mechanisms. In fact, cystathionine beta synthase deficiency had no effect on glutathione peroxidase, glutathione reductase and glutathione S-transferase activities. However, we found a 50% increase in hepatic catalase activity without any variation of expression. These findings demonstrate that cystathionine beta synthase deficiency initiates redox disequilibrium in the liver. However, the activation of catalase attenuates oxidative impairments.

In higher plants, microtubules (MTs) are assembled in distinctive arrays in the absence of a defined organizing center. Three MT nucleation sites have been described: the nuclear surface, the cell cortex and cortical MT branch points. The Arabidopsis thaliana (At) genome contains putative orthologues encoding all the components of characterized mammalian nucleation complexes: gamma-tubulin and gamma-tubulin complex proteins GCP2 to GCP6. We have cloned the cDNA encoding AtGCP2, and show that gamma-tubulin, AtGCP2 and AtGCP3 are part of the same tandem affinity-purified complex and are present in a large membrane-associated complex. In addition, small soluble gamma-tubulin complexes of the size expected for a gamma-tubulin core complex are recruited to isolated nuclei. Using immunogold labelling, AtGCP3 is localized to both the nuclear envelope (NE) and the plasma membrane. To identify domains that could play a role in targeting complexes to these nucleation sites, truncated AtGCP2- and AtGCP3-green fluorescent protein fusion proteins were expressed in BY-2 cells. Several domains from AtGCP2 and AtGCP3 are capable of targeting fusions to the NE. We propose that regulated recruitment of soluble gamma-tubulin-containing complexes is responsible for nucleation at dispersed sites in plant cells and contributes to the formation and organization of the various MT arrays.

Cilia exert critical functions in numerous organisms, including that of cell motility, fluid transport and protozoan locomotion. Defects in this organelle can lead to lethal pathologies in humans, including primary ciliary dyskinesia. An understanding of the cilia formation process would lead to better characterization of defects involved in such pathologies. In the present study, we identified a gene encoding a novel human protein, BCAP for Basal body Centriole-Associated Protein, which shares homologies with a previously described protein, Outer Dense Fiber 2 (ODF2). ODF2, a major component of the sperm tail cytoskeleton, is required for the formation of mother centriole distal/subdistal appendages and the generation of primary cilia. Here, we show that the bcap gene contains 18 alternatively spliced exons and encodes five different isoforms, three long and two short ones. BCAP is preferentially expressed in cilia/flagella containing tissues. Moreover, its expression is correlated with cilia formation during mucociliary differentiation of human nasal epithelial cells. Using immunofluorescence analyses, BCAP was localized within basal bodies of ciliated cells and within centrioles of proliferating cells. In light of the several spliced isoforms of BCAP and the particular localization of the protein, BCAP isoforms could play distinct roles in cilia and in centrosomes.

Objective: To determine whether our health care employees were undergoing mammography screening according to American Cancer Society guidelines and to determine whether aggressive outreach, education and streamlining of mammography scheduling could improve compliance., Study Design: All female employees at North Shore University Hospital (NSUH) and several other health system facilities (SF) were sent mailings to their homes that included breast health education and mammography screening guidelines, a questionnaire regarding their own mammography screening history and the opportunity to have their mammography screening scheduled by the Mammography Screening Employee Inreach Program (MSEIP) coordinator., Results: Of the approximately 2,700 female employees aged 40 and over at NSUH and SF, 2,235 (82.7%) responded to the questionnaire, and 1,455 had a mammogram done via the MSEIP. Of the 1,455, 43% either were overdue for a mammogram or had never had one. During a second year of the MSEIP at NSUH and SF, an additional 1,706 mammograms were done., Conclusion: People employed in health care jobs do not necessarily avail themselves of appropriate health care screening. An aggressive program that utilized education, outreach and assistance with scheduling was effective in increasing compliance with mammography screening.

Cilia are specialized organelles that exert critical functions in numerous organisms, including that of cell motility, fluid transport and protozoan locomotion. Ciliary architecture and function strictly depend on basal body formation, migration and axoneme elongation. Numerous ultrastructural studies have been undertaken in different species to elucidate the process of ciliogenesis. Recent analyses have led to identification of genes specifically expressed in ciliated organisms, but most proteins involved in ciliogenesis remain uncharacterized. Using human nasal epithelial cells capable of ciliary differentiation in vitro, differential display was carried out to identify new proteins associated with ciliogenesis. We isolated a new gene, ICIS-1 (Involved in CIlia Stability-1), upregulated during mucociliary differentiation. This gene is localized within the TGF-beta1 promoter and is ubiquitously expressed in human tissues. Functional analyses of gene expression inhibition by RNA interference in Paramecium tetraurelia indicated that the ICIS-1 homologue interfered with cilia stability or formation. These findings demonstrate that ICIS-1 is a new protein associated with ciliated cells and potentially related to cilia stability.

Cytochromes P450 monooxygenases from the CYP98 family catalyze the meta-hydroxylation step in the phenylpropanoid biosynthetic pathway. The ref8 Arabidopsis (Arabidopsis thaliana) mutant, with a point mutation in the CYP98A3 gene, was previously described to show developmental defects, changes in lignin composition, and lack of soluble sinapoyl esters. We isolated a T-DNA insertion mutant in CYP98A3 and show that this mutation leads to a more drastic inhibition of plant development and inhibition of cell growth. Similar to the ref8 mutant, the insertion mutant has reduced lignin content, with stem lignin essentially made of p-hydroxyphenyl units and trace amounts of guaiacyl and syringyl units. However, its roots display an ectopic lignification and a substantial proportion of guaiacyl and syringyl units, suggesting the occurrence of an alternative CYP98A3-independent meta-hydroxylation mechanism active mainly in the roots. Relative to the control, mutant plantlets produce very low amounts of sinapoyl esters, but accumulate flavonol glycosides. Reduced cell growth seems correlated with alterations in the abundance of cell wall polysaccharides, in particular decrease in crystalline cellulose, and profound modifications in gene expression and homeostasis reminiscent of a stress response. CYP98A3 thus constitutes a critical bottleneck in the phenylpropanoid pathway and in the synthesis of compounds controlling plant development. CYP98A3 cosuppressed lines show a gradation of developmental defects and changes in lignin content (40% reduction) and structure (prominent frequency of p-hydroxyphenyl units), but content in foliar sinapoyl esters is similar to the control. The purple coloration of their leaves is correlated to the accumulation of sinapoylated anthocyanins.

Transport of the viral genome into the nucleus is an obligatory step in the replication cycle of plant pararetro- and geminiviruses. In both these virus types, the multifunctional coat protein (CP) is thought to be involved in this process. Here, a green fluorescent protein tagging approach was used to demonstrate nuclear import of the CPs of Rice tungro bacilliform virus (RTBV) and Mungbean yellow mosaic virus--Vigna (MYMV) in Nicotiana plumbaginifolia protoplasts. In both cases, at least two nuclear localization signals (NLSs) were identified and characterized. The NLSs of RTBV CP are located within both N- and C-terminal regions (residues 479KRPK/497KRK and 744KRK/758RRK), and those of MYMV CP within the N-terminal part (residues 3KR and 41KRRR). The MYMV and RTBV CP NLSs resemble classic mono- and bipartite NLSs, respectively. However, the N-terminal MYMV CP NLS and both RTBV CP NLSs show peculiarities in the number and position of basic residues. In vitro pull-down assays revealed interaction of RTBV and MYMV CPs with the nuclear import factor importin alpha, suggesting that both CPs are imported into the nucleus via an importin alpha-dependent pathway. The possibility that this pathway could serve for docking of virions to the nucleus is discussed.

Millions of American girls and women have been drawn to smoking by an industry that has been clearly and systematically targeting women of all ages and life circumstances. Tobacco marketing strategies skillfully link cigarette use to typical female values. Biologically speaking, women are especially vulnerable to the legion of health problems of tobacco use. Smoking is a critical hazard for women in their reproductive years, particularly when they are pregnant.

Breast cancer is reported to occur in from 1 in 3000 to 1 in 10,000 pregnancies worldwide. In the United States, 10% to 20% of breast cancers occur in women of childbearing age. Diagnosis and treatment of breast cancer during pregnancy present many additional challenges.

It is estimated that more than 200 million women smoke worldwide. It is also estimated that if current smoking rates among men and women persist worldwide, by 2025, 9% of the world's deaths and disabilities will be related to tobacco use. Smoking is associated with an increased risk of cardiovascular disease, cancer (lung, cervix, pancreas, kidney, bladder, esophagus, pharynx), pulmonary disorders, cerebrovascular disease, and many other health risks. Smoking also adversely affects fertility and reproductive outcomes. Although a greater percentage of women smoke in developed than in developing countries, women in developing countries appear to be initiating smoking habits in increasing numbers. If current trends persist, the adverse effects of smoking on women's health clearly will continue to escalate.

Each year more than 600000 women have deaths associated with cigarette smoking. In addition, cigarette smoking is associated with a wide array of morbidities (such as osteoporosis, cardiovascular disease, and adverse pregnancy outcomes). Two hundred million women smoke worldwide, and this number appears to be rising, particularly in developing countries. Obstetrician-gynecologists can play a role in reducing morbidity and mortality from cigarette smoking by educating women about the dangers, advising them not to smoke, and assisting those who do smoke to quit.

Cancer is the second leading cause of death for women in the United States. In 1998, there were approximately 600,000 new cases of cancer and 270,000 cancer deaths in U.S. women. Many of these cancer deaths are preventable. In 1987, lung cancer replaced breast cancer as the leading cause of cancer deaths in U.S. females. Cancer of the lung and bronchus was responsible for 25% of cancer deaths in U.S. women last year, killing approximately 67,000 women. It has been estimated that almost 90% of lung cancer deaths are associated with cigarette smoking and, therefore, are preventable. Some approaches to smoking cessation are reviewed. Breast cancer was responsible for 30% of cancers (178,700 women) and 16% of cancer deaths (43,500 women). Strategies for early detection and, possibly, for prevention are reviewed. The third most common cancer and the third most common cause of cancer deaths in U.S. women is colorectal cancer. Strategies for prevention and early detection are addressed. Cancer of the reproductive system affects approximately 80,000 U.S. women each year and results in approximately 27,000 deaths. Current screening capabilities, as well as some of the limitations of our current screening technologies, are addressed. The healthcare team, working in concert with our patients, government, and the health insurance industry, should be able to substantially reduce both the incidence of and mortality from cancer in women.

In obstetrics and gynecology, more than 60% of current residents and 36% of medical school faculty are women. However, fewer than 10% of professors and only a small number of department chairs and organizational leaders are women. This paper reviews the data on the growing number of women in obstetrics and gynecology, and in medicine in general, during the past 25 years. It reviews some of the challenges that women have faced and some strategies to enhance equitable opportunities.

Objective: To test the hypothesis that the Residency Review Committee program requirements for obstetrics and gynecology residencies, when properly followed, will result in residents being educated in preventive and primary ambulatory health care for women during their residency training program as specialists in obstetrics and gynecology., Methods: The 60 requisite residency training competencies identified as essential to educate generalist physicians, and viewed by some educators as a benchmarking standard, each were evaluated to determine whether residents in obstetrics and gynecology are now being educated in each of these areas. The answer was considered affirmative if any of the following pertained: 1) the Residency Review Committee program requirements indicate that the competency "must" or "should" be taught, 2) the Residency Review Committee requests numerical verification related to the competency on the accreditation review application, or 3) by virtue of a specific rotation required by the Residency Review Committee it can be assumed that the resident will be educated in the competency. To make our assessment, we identified and listed the section of the Residency Review Committee for Obstetrics-Gynecology program requirements, which, when properly followed, would result in education in the particular competency., Results: Fifty-seven of the 60 competencies were considered applicable to obstetrician-gynecologists (care of infants, care of children, and infant/child preventive care were not), and residents in obstetrics and gynecology were found to be educated in 54 (95%)., Conclusion: During their residency training programs as specialists in obstetrics and gynecology, residents are being educated to be able to be providers of preventive and ambulatory primary health care for women.

Objective: The objective of this study was to determine whether hormone production by postmenopausal ovaries containing benign ovarian tumors differed from that of normal postmenopausal ovaries., Study Design: The sera of 32 postmenopausal patients were assayed before and after bilateral oophorectomy for estrone, 17 beta-estradiol, androstenedione, testosterone, and dehydroepiandrosterone sulfate. The data from all patients as a group were analyzed, followed by analysis of the data from 15 patients with normal ovaries separately from the remaining 17 patients who had nonfunctioning, benign ovarian tumors., Results: For patients with benign ovarian tumors there was a statistically significant drop in estrone (from a presurgical level of 55.8 +/- 46.3 pg/ml to a postoperative level of 29.9 +/- 10.2 pg/ml, p < 0.03) and 17 beta-estradiol (from 18.6 +/- 14.1 pg/ml preoperatively to 9.8 +/- 3.8 pg/ml postoperatively, p < 0.02). For postmenopausal woman with normal ovaries there was no significant drop in estrone or 17 beta-estradiol after bilateral oophorectomy. There was a statistically significant drop in testosterone and androstenedione after bilateral oophorectomy both for women with normal ovaries and for those with benign tumors. No significant differences in dehydroepiandrosterone sulfate were noted in either group., Conclusions: These data suggest that, although normal postmenopausal ovaries have not been demonstrated to secrete clinically significant amounts of estrogen, those that contain benign ovarian tumors do secrete small but significant amounts of estrone and 17 beta-estradiol. Both tumor-containing and normal ovaries secrete the androgens androstenedione and testosterone, this secretion not being significantly influenced by the presence of a tumor.

Objective: To determine whether large-loop excision of the transformation zone (LLETZ) can be used in our population to treat patients in a routine colposcopy clinic without diminishing the capability to see the large number of patients who require care, and to compare these results to our previous experience with laser surgery., Methods: Patients found to have squamous intraepithelial lesions on colposcopically directed biopsies were offered treatment with LLETZ during counseling regarding their biopsy findings. Procedures included "ablation equivalents" and "cone biopsy equivalents" using local anesthesia. Follow-up examinations were used to determine cure and included cytology, colposcopy, and directed biopsies when indicated. The cure rate was compared to our previous experience with laser surgery., Results: Two hundred thirty-six patients were treated in the colposcopy clinic without diminishing the capability to see all patients requiring care. Complications were few. The mean (+/- standard deviation) follow-up period was 50.7 +/- 25.3 weeks. The overall cure rate of 91.3% (95% confidence interval [CI] 87.1-95.5) was not influenced by the severity of the disease, but positive endocervical margins significantly lowered the cure rate to 69.2% compared with those who had negative margins. Before this series, only 73.1% of our patients scheduled for laser surgery returned for treatment. Assuming a 90% cure rate among those who returned, this means that the actual cure rate was only 65.8%. The likelihood of cure was 1.37 times greater (95% CI 1.27-1.52; P < .0001) using LLETZ in the clinic at the time the patient was counseled regarding her biopsy findings than using laser at a later date. In 33 patients, the LLETZ specimen showed no evidence of disease. The relative risk of negative histology was 3.31 (95% CI 1.78-6.13; P < .001) when LLETZ was done for a discrepancy between cytology and histology as opposed to any other indication. Cancer was found on the LLETZ specimen in four patients (two microinvasive, two frankly invasive), but was not suspected preoperatively in any of the patients., Conclusion: In our inner-city clinic, treatment with LLETZ at the time the patient was counseled regarding her biopsy findings improved the actual cure rate. The LLETZ procedure can be done safely in a clinic setting without diminishing the capability to care for a large number of patients.

Polycystic ovarian disease is both a hyperestrogenic and a hyperandrogenic syndrome, and all studies have shown that hyperestrogenemia is the result of an elevation of estrone with plasma estradiol levels in the normal follicular range. Because a literature search failed to reveal any report of polycystic ovarian disease with significantly elevated estradiol levels, we report a case in which the plasma estradiol was so massively elevated as to mimic an estrogen-producing neoplasm. This case also suggests that although polycystic ovarian disease is a very rare cause of such excessive estradiol production, it should be included in the differential diagnosis of estrogen-producing neoplasms.

Objectives: The Association of Professors of Gynecology and Obstetrics and the Council on Resident Education in Obstetrics and Gynecology have proposed a fourth-year medical school curriculum for a student interested in pursuing a residency in obstetrics and gynecology., Study Design: Faculty members and residents in North Carolina, Illinois, and Michigan were surveyed as to the ideal curriculum that they would recommend for fourth-year students. The committee members representing the Council on Resident Education in Obstetrics and Gynecology and the Association of Professors of Gynecology and Obstetrics then reviewed these surveys and proposed a final curriculum., Results: A core curriculum of general medicine as an acting internship, an intensive care unit rotation, neonatology, and emergency medicine was recommended. Additional courses strongly considered were ambulatory obstetrics-gynecology, acting internship in obstetrics-gynecology, endocrinology, and general surgery., Conclusion: The committee recommends a curriculum that is broad and balanced in general medical education.

Laparoscopic surgery for ovarian lesions has the potential for an enormous positive impact on the care that we are able to provide for our patients. Technologic advances and the proliferation of new equipment have occurred at a rapid rate. However, the criteria for the safe and appropriate use of endoscopic surgery have not been developed fully or followed by all surgeons, and there is some evidence that a lack of adherence to carefully considered protocols has the potential to have an adverse impact on women with ovarian lesions. McDonough made a thoughtful presentation on the need for technology assessment in the reproductive sciences, focusing on the fact that there has been a rapid diffusion of technology without prior broad-based critical peer review and stressing the need to have scientific technology assessment. Pitkin, in his editorial on operative laparoscopy, questioned whether we are evaluating therapeutic appropriateness properly as a separate and independent issue from technical feasibility. Obviously, it would be preferable to have had more controlled prospective data on the relative merits and risks of the use of the laparoscope for resection of ovarian cysts before the widespread use of this technology. However, there are several situations in which it currently appears to have important clinical utility for the patient. In addition, it already is being used widely, and it would be inappropriate to withhold this method from our patients. However, it should be urged that careful prospective evaluation be undertaken so that both the scientific community and the public are able to have an accurate understanding of the risks, benefits, indications, and contraindications of this technology. In addition, when a surgeon is considering pelviscopic surgery for an ovarian mass, it is essential that this be done with strict adherence to carefully constructed preoperative criteria and intraoperative protocols.

We sought to determine the prevalence of human immunodeficiency virus (HIV) infection in a population of women with human papillomavirus (HPV)-related diseases attending a colposcopy clinic who had no other CDC-defined risk factors for HIV. Study patients included all new patients attending our colposcopy clinic who were found to have histologic evidence of condyloma or cervical intraepithelial neoplasia. Those patients not already known to be HIV-positive were offered testing for HIV. Demographic information was obtained on all patients. Results were compared to data from anonymous testing of our own obstetrical population. One hundred forty of 208 women (67.3%) were either previously known to be HIV-positive or agreed to be tested. Sixteen (11.4%) were HIV-positive. Eight of the HIV-positive women were not previously known to be HIV-positive and 6 of the 8 had no definable risk factors for HIV infection. This is 4.6% of the women not already known to have a CDC-defined risk factor for HIV. The rate of HIV infection in our obstetrical population is 1.6%. In women without other definable risks for HIV infection and who had HPV-related disease the relative risk of HIV infection in our population was 2.94 (95% confidence interval 1.21-6.94; P < 0.031). In areas where HIV is endemic there is a high prevalence of HIV infection in women with HPV-related disease. Even in women without another definable risk factor for HIV, HPV-related disease may serve as a marker for an increased risk of HIV infection in this population.

Objective: Our goal was to determine the rate of attrition from obstetrics and gynecology residency programs., Study Design: The Council on Resident Education in Obstetrics and Gynecology sent questionnaires to all 295 obstetrics and gynecology residency program directors in the United States and Canada. These programs represent 4306 postgraduate-year 1 through 4 (or 5) resident positions each year. The program directors were asked the number of residents who left voluntarily or were dismissed in a 2-year period and the reasons they left., Results: In a 2-year period 299 residents left or were dismissed (6.94% over 2 years, or 3.47% per year). Only 88 (1% per year) left specifically because they decided they preferred a different discipline., Conclusion: The rate of attrition from obstetrics and gynecology residency programs is not excessively high.

A comparison was made of the ACTH stimulation test in the proliferative and luteal phases of the menstrual cycle in 13 subjects, each of whom served as her own control. The test yielded a 53.9% false-positive rate in the luteal phase. The study, therefore, clearly shows that from a practical, clinical point of view, the test is completely unreliable in the luteal phase, and is only valid when carried out in the follicular phase of the cycle.

The safety and efficacy of patient-controlled analgesia for the long-term control of cancer pain was tested prospectively. Respiratory rates, mental status, and pain relief were recorded at baseline and compared with those during the study period. Patients had a lower analgesic demand (i.e., self-administered less morphine during the nighttime); specifically, dosing declined 48% from the daytime level. Respiratory rates did not change appreciably during the study and no cases of significant respiratory depression were encountered. Patients self-administered sufficient morphine to produce adequate but not complete pain relief in almost all trials. Pain relief was safely achieved by both intravenous and subcutaneous routes of administration in both the inpatient and outpatient settings. Mean 24-h morphine use stayed relatively constant even for patients receiving more than 2 weeks of treatment. In conclusion, patient-controlled analgesia is effective and safe therapy for the long-term control of severe cancer pain.

Obstetrics and gynecology residency programs have traditionally involved long hours in the hospital. In recent years, in an attempt to determine whether work hours could be reduced while at least maintaining resident education and patient care, many program directors have instituted night float systems. In New York State, these systems must adhere to rigid hospital code requirements (limiting total hours worked and with specific mandates regarding time away from the hospital); in other areas, these requirements are not as limiting. At the request of the Council on Resident Education in Obstetrics and Gynecology, residency program directors and residents in the United States and Canada were sent a survey regarding whether they had a night float program, how it was structured, and what changes it was perceived to have caused. Responses were received from 193 program directors (65%) and 302 residents. Major differences were noted in the structure of the programs within New York state compared with those outside the state. In New York, 63% of the programs had residents in all 4 years participating in the night float; this was true for only 10% of the programs outside New York. In New York state, the programs were required to adhere to state hospital code requirements limiting hours on duty and mandating the specifics of time off, whereas the programs outside New York did not necessarily adhere to these restrictive requirements. Twelve characteristics were evaluated regarding changes that were perceived to have occurred as a result of the night float program.(ABSTRACT TRUNCATED AT 250 WORDS)

A comprehensive program was founded in 1982 to provide adolescents with prenatal and family planning care. The program's impact through its first five years of operation on medical aspects of pregnancy course and fetal outcome will be the subject of a separate report. This study examines subsequent maternal and infant health of the patients attending the program compared to a control group. Four hundred ninety-eight adolescents and their newborns attending the program's mother-baby family planning clinic from 1982 to 1989 (subject group) were compared to ninety-one adolescents and their newborns receiving postpartum family planning and pediatrics clinics from 1980 through 1989 (control group). Seventy-five percent of the subject group regularly attended mother-baby clinic, compared to 18% of the control group attending family planning and pediatric clinics (P less than or equal to .0001). The subject group experienced less maternal and infant morbidity, greater school attendance, graduation, employment, and contraceptive use than the control group (P less than or equal to .0001). Many parameters improved with each program year indicating continued wide acceptance of our program by area adolescents.

One hundred fifty-six members and candidate members of the Society of Gynecologic Oncologists responded to a survey concerning the "laparoscopic management of ovarian neoplasms subsequently found to be malignant." Twenty-nine responders (19%) reported a total of 42 cases of ovarian malignancy. The laparoscopic procedure was aborted or the cyst was aspirated in 38% of the cases, and partial or complete excisions were attempted in 33 and 29%, respectively. The characteristics of the masses were as follows: less than 8 cm 67%, cystic 62%, unilocular 48%, and unilateral 81%. All four "benign" characteristics were present in 31% of the cases found to be malignant, and three of four characteristics were present in 24%. Laparotomy was performed at the time of laparoscopy in 17% of cases, after laparoscopy in 71% with an average interval of 4.8 weeks, and not at all in 12%. Fifty-seven percent of the cases were invasive epithelial malignancies, whereas 29% were tumors of low malignant potential. At least 50% of the patients had stages II-IV. We conclude that attempted laparoscopic excisions of adnexal masses that are subsequently found to be malignant are not uncommon, and that the presence of so-called "benign" characteristics does not preclude malignancy. Attempts at partial or complete excision are common, as are delays in subsequent definitive surgery. The stage of disease is often advanced, and all histologic types of malignancy are encountered. We advocate careful evaluation of this practice, with development of strict guidelines to ensure optimal patient care.

In 1988, New York became the first state to implement regulatory measures limiting housestaff work hours. Because restrictions on residents' work hours will have such profound and far-reaching implications for how obstetrics and gynecology residencies are conducted, the Council on Resident Education in Obstetrics and Gynecology (CREOG) requested that a survey be conducted to solicit information from program directors of U.S. obstetrics and gynecology residencies who had already begun to alter their call schedules. Two hundred and ninety-six programs were contacted, and representatives of those that had implemented changes were requested to respond. Eighty-two responses were received; 26 of these contained information that could be collated. From these 26 responses the authors have structured a prototypic call schedule and presented its application. A key feature necessary to implement the new type of schedule is the use of a night float system. It is concluded that changes can and will be made by obstetrics and gynecology residencies. Creative scheduling, as described in this article, is essential and will facilitate the task; however, the current standards of education and patient care will be difficult to maintain without additional economic and human resources.

The value of serum beta-hCG measurement in the diagnosis of ectopic pregnancy is well established, and there have been recent studies on the use of serum progesterone levels. However, we have been unable to find any reports on the potential application of serum estradiol (E2) assays in the diagnosis of ectopic pregnancy. We therefore concurrently measured serum E2, progesterone, and beta-hCG in 100 women with ectopic pregnancies, as well as in 69 controls with normal intrauterine pregnancies and 36 women with threatened abortion. The mean (+/- standard deviation) E2 levels for ectopic-pregnancy patients, the normal controls, and the women with threatened abortion were 281.1 +/- 115.6, 788.2 +/- 45.5, and 788.8 +/- 40.6 pg/mL, respectively; the mean levels in the ectopic group were significantly different (P less than .0001) from those of the other two groups. All but one of the ectopic pregnancies had values below 650 pg/mL for E2 and 23 ng/mL for progesterone, and all but one of the normal intrauterine pregnancies had values above these levels. Our data suggest that the addition of the estradiol assay, with or without progesterone, to the early evaluation of patients suspected of having an ectopic pregnancy may be helpful in diagnosis.

We report the case of a patient who was successfully treated with a long-acting GnRH agonist for pulmonary endometriosis. This 28-year-old woman had symptomatic pleural endometriosis, documented by biopsies, as well as symptomatic pelvic endometriosis. Two surgical procedures, consisting of excision of pleural endometriotic tissue and partial pleurectomies, failed to relieve her chest symptoms. Little relief was achieved with pseudopregnancy treatment. Satisfactory symptomatic improvement was obtained with danazol, but this medication had to be discontinued because of severe side effects. Trial of a GnRH agonist, leuprolide acetate, achieved complete remission of her chest symptoms; in addition, the patient became pregnant immediately after cessation of therapy. Gonadotropin-releasing hormone agonist therapy may be an important therapeutic alternative for women with pulmonary endometriosis who cannot tolerate danazol treatment and in whom surgical therapy fails to relieve the chest symptoms.

Thirty-eight postmenopausal women and one with pituitary amenorrhea underwent cervical laser surgery. The cervical canal of each patient was patent 2 weeks later. Eighteen of the 30 women reevaluated 4-48 months later had total obliteration of the cervical canal. In some patients, the cervix was obliterated and flush with the vaginal vault so that it could not be identified. Five other patients had severe cervical stenosis. Factors contributing to cervical os obliteration appeared to be the depth of the cervical defect, the lack of the physical action of blood passing through the cervical canal, and the hypoestrogenic state. This outcome makes continued adequate cytologic and colposcopic surveillance of these patients impossible and leaves them at risk for developing unrecognized cervical and endometrial disease.

The charts of 500 sequential perimenopausal patients were reviewed and their alterations in menstrual flow were characterized as fitting one of three patterns. Sixty-two patients (12%) had sudden amenorrhea, 347 (70%) had oligomenorrhea and/or hypomenorrhea, and 91 (18%) had menorrhagia, metrorrhagia, and/or hypermenorrhea. All of the patients in the menorrhagia/metrorrhagia and/or hypermenorrhea group had histologic evaluations to rule out the presence of premalignant or malignant disease; 17 women (19%) in this group had premalignant or malignant findings. In addition, 4 of the 5 women with malignancies had intermenstrual bleeding. Of the 9 women in the study with intermenstrual bleeding, 4 had invasive cancer and 2 had endometrial hyperplasia. This study documents and highlights the high incidence of premalignant and malignant findings in perimenopausal patients with bleeding patterns other than amenorrhea or oligomenorrhea/hypomenorrhea, and underscores the need for perimenopausal women with menorrhagia, metrorrhagia, and/or hypermenorrhea to undergo thorough evaluation.

The uterine adenofibroma is a form of mixed mesodermal tumor in which both epithelial and stromal components are benign. A case is presented in which a patient with this lesion was treated with excision of the tumor rather than hysterectomy. Two years later, she developed a recurrence of the lesion, which was again treated with wide local excision. A hysterectomy was done 9 months later, and a polypoid lesion was noted in the fundus of the uterus which on histologic examination was thought to be potentially either an adenofibroma precursor or a maturation of the previous cellular adenofibroma. Since uterine adenofibroma usually occurs in older patients, the standard treatment has been hysterectomy. Because of this patient's age, she was initially treated with a more conservative approach. However, despite the fact that the uterine adenofibroma is a benign lesion, it has an obvious potential to recur when treated conservatively.

There is evidence to suggest that the human immunodeficiency virus (HIV) can be transmitted from mothers to their uninfected offspring via infected breast milk. The quantitative risk of transmission by this route is unknown. We must urge all individuals at risk for HIV infection to be tested. In the United States, it is recommended that women who are known to be HIV-positive be discouraged from breast-feeding. In certain regions of the United States, such as high-risk areas of New York City, HIV infection may be present in more than 3% of reproductive-age women, most of whom do not know they are infected. Women who either live in areas of the United States in which acquired immunodeficiency syndrome (AIDS) is endemic or are living in other parts of the country and are at high risk for AIDS, yet refuse to be tested, should be counseled regarding the potential for transmission of the virus to uninfected offspring via breast-feeding.

Psammoma bodies have never previously been noted in conjunction with cervical cancer. This paper presents a case of endocervical adenocarcinoma with psammoma bodies. Malignant and benign lesions which have previously been associated with psammoma bodies are reviewed. The proposed mechanisms of psammoma body formation are considered. The appropriate action upon finding psammoma bodies on cervicovaginal cytology is discussed.