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Allocation of the thoracic imaginal primordia in the Drosophila embryo.

The primordia of the thoracic imaginal discs of the Drosophila embryo originate as groups of cells spanning the parasegment boundary. We present evidence that the thoracic imaginal primordia are allocated in response to signals from the wingless (wg) and decapentaplegic (dpp) gene products. Rows of cells that express wg intersect rows of cells that express dpp to form a ladder-like pattern in the ectoderm of the germ band extended embryo. The imaginal primordia originate as groups of cells which lie near these intersection points. We have used a molecular probe derived from the Distal-less (Dll) gene to show that this population contains progenitor cells for both the dorsal (i.e. wing) and ventral (i.e. leg) discs. Although we show that Dll function is not required for allocation of imaginal cells, activation of an early Dll enhancer may serve as a molecular marker for allocation. A group of cells, which includes the imaginal progenitors, activate this enhancer in response to intercellular signals from wg and perhaps from dpp. We have used a conditional allele of wg to show that wg function is transiently required for both allocation of the imaginal primordia and for initiation of Dll expression in these cells during the brief interval when wg and dpp form the ladder-like pattern. Allocation of the imaginal primordium and activation of Dll expression appear to be parallel responses to a single set of positional cues.

Animals↗

Combined activities of Gurken and decapentaplegic specify dorsal chorion structures of the Drosophila egg.

During Drosophila oogenesis Gurken, associated with the oocyte nucleus, activates the Drosophila EGF receptor in the follicular epithelium. Gurken first specifies posterior follicle cells, which in turn signal back to the oocyte to induce the migration of the oocyte nucleus from a posterior to an anterior-dorsal position. Here, Gurken signals again to specify dorsal follicle cells, which give rise to dorsal chorion structures including the dorsal appendages. If Gurken signaling is delayed and starts after stage 6 of oogenesis the nucleus remains at the posterior pole of the oocyte. Eggs develop with a posterior ring of dorsal appendage material that is produced by main-body follicle cells expressing the gene Broad-Complex. They encircle terminal follicle cells expressing variable amounts of the TGFbeta homologue, decapentaplegic. By ectopically expressing decapentaplegic and clonal analysis with Mothers against dpp we show that Decapentaplegic signaling is required for Broad-Complex expression. Thus, the specification and positioning of dorsal appendages along the anterior-posterior axis depends on the intersection of both Gurken and Decapentaplegic signaling. This intersection also induces rhomboid expression and thereby initiates the positive feedback loop of EGF receptor activation, which positions the dorsal appendages along the dorsal-ventral egg axis.

Animals↗

The membrane skeleton of acetylcholine receptor domains in rat myotubes contains antiparallel homodimers of beta-spectrin in filaments quantitatively resembling those of erythrocytes.

I used immunogold labeling and quick-freeze, deep-etch, rotary replication to characterize the membrane skeleton at regions with high concentrations of acetylcholine receptor domains in receptor clusters of cultured rat muscle cells. This membrane skeleton consists of a network of filaments closely applied to the cytoplasmic membrane surface. The filaments are specifically decorated by immunogold labeling with a monoclonal antibody, VIIF7, that recognizes an isoform of beta-spectrin colocalizing with acetylcholine receptors. The filaments are 32 +/- 11 nm in length and three to four filaments (average 3.1-3.3) join at each intersection to form the network. These parameters are nearly identical to those reported previously for the membrane skeleton of erythrocytes. Depending on the amount of platinum coating, filament diameters range from 9 to 11 nm in diameter, and are 1.4 nm larger on average than spectrin filaments of erythrocytes replicated at the same time. Filaments are decorated with gold particles close to one end, consistent with the location of the epitope recognized by the monoclonal antibody. Computer modeling shows that all filament intersections in the membrane skeletal network are equally capable of being labeled by the monoclonal antibody. This pattern of labeling is consistent with a network containing antiparallel homodimers of beta-spectrin.

Animals↗

Desmosomes in hamster cheek pouch epithelium: their quantitative characterization during epithelial differentiation.

Desmosomes in stratified squamous epithelia appear to exhibit quantitative alterations during differentiation. In this work we use stereological and other morphometric methods to quantify these structures in epithelial cells from defined basal, spinous and granular strata. Hamster cheek pouch mucosa from five animals was processed for electron microscopy using strictly standardized techniques and a stratified random sampling procedure was used to obtain micrographs of cells from basal, spinous and granular layers. Stereological intersection counting techniques were used to determine for each layer the relative surface area of plasma membrane occupied by desmosomes (Ss), the number of desmosomes per unit surface area of plasma membrane (Ns), the mean individual desmosomal diameter (delta) and the mean individual desmosomal surface area (s). In addition, estimates of nuclear volume were obtained by direct measurement of nuclear profiles and volume-to-surface ratios were obtained by a combination of point and intersection counting, which enabled estimates for the volume (Vcell) and plasma membrane surface area (SPM) of the 'average' cell within each stratum to be acquired. Using this information, it was then possible to calculate both the total surface area (S) and the number (N) of desmosomes on the plasma membranes of average cells. The parameters Ss and Ns showed progressive increases between basal and granular layers, whereas values for delta and s were lower in granular cells when compared with basal and spinous cells. The parameters Vcell, SPM, S and N all increased progressively and significantly during differentiation. Between basal and granular layers, the mean cell volume and surface area had each increased approximately threefold, whereas the surface area and number of desmosomes on the average cell plasma membrane had increased approximately seven- and eleven-fold, respectively. Granular cells thus possess more numerous desmosomes, which occupy a greater proportion of the plasma membrane area but which are individually smaller, when compared with basal and spinous layers.

Animals↗

Localization of the sentinel node of the upper outer breast quadrant in the axillary quadrants.

BACKGROUND: Sentinel node (SN) biopsy is associated with much less morbidity than axillary dissection. In patients with early breast cancer, lymphatic mapping and SN biopsy accurately stage the axillary nodes. Both currently available lymphatic mapping agents, radiocolloid and blue dye, have some limitations that may make perioperative or preoperative SN identification difficult. In such cases, exact knowledge of the topography of the axilla and the most probable location of the SN may be crucial. METHODS: In 12 fresh female cadavers with no history of breast carcinoma, injections of patent blue dye were used to visualize the SNs in the axillary quadrants and their lymphatic collectors from the upper outer quadrant of the breast, which is the most common location of breast cancer. The axilla was divided into quadrants with regard to the intersection of the thoracoepigastric vein and the third intercostobrachial nerve. RESULTS: All SNs were located within a circle of 2-cm radius of this intersection in the fatty tissue at the clavipectoral fascia. In most cases, the SN was located in the fatty tissue near the clavipectoral fascia in the lower ventral quadrant of the axilla (n = 14, 58%). In seven cases (29%), the SN was located in the upper ventral quadrant, in two cases (8%) in the upper dorsal quadrant, and in one case in the lower dorsal quadrant. CONCLUSIONS: The results of this anatomical study may facilitate SN biopsy in patients with breast cancer.

Aged↗

Evaluation of the cortical structure in high resolution CT images of lumbar vertebrae by analysing low bone mineral density clusters and cortical profiles.

The structural classification of trabecular bone is of considerable clinical importance for the diagnosis of osteoporosis. Assessment of the cortical bone mineral density (BMD) and analysis of cortical structure is necessary because the cortex is also affected by osteoporosis. We describe a method for the automatic classification of the cortex from its structure on high resolution (HR) CT images. The method is based on the distribution of areas with low BMD in the cortex. Two different approaches are presented; one uses the grey scale profile of the cortical ridge, and the other one evaluates the distribution of connected regions (clusters) of low BMD. i.e. areas that lie below a certain threshold value. In HRCT images from 22 lumbar vertebrae, the number of intersections of the cortical intensity profile with a horizontal line was determined at various positions of this threshold line. At a certain position, there was a local maximum in the number of intersections which was 38% higher in the osteoporotic cases. The maximum shows a moderate correlation with the cortical BMD of rni = -0.72 (p < 0.0001). The number nc of connected regions (clusters) with pixel values below a certain threshold value was determined in relation to the threshold value T. The resulting function nc(T) shows a relative maximum slightly below the average grey scale value of the respective image. The curve depends on the degree of osteoporosis: the height of the maximum (i.e. the maximal number of clusters ncmax) allows distinction to be made between different degrees of osteoporosis, and ncmax shows a correlation with the cortical BMD of rnc = -0.84 (p < 0.0001).

Aged↗

[Baseline study of sleep electroencephalography--daily variation and individual variation].

The aims of the present study were to observe the daily habituation to night sleep in a laboratory environment and to make clear the daily and individual sleep variations by using polygraph parameters, including electroencephalography (EEG). Sleep EEG records were obtained from a subject who slept ten successive nights, and from six subjects who each slept one night in the laboratory. The parameters used were as follows: sleep stage %, sleep latency (SL), REM latency (RL), number of stage shifts, subjective sleep, integral EMG, and slope (a) and intersect (b) of a regression equation used to estimate the sleep depth against sleep time. Stage WAKE and SL, slope (a), intersect (b) and the mean depth of sleep were found to become stable from the fifth night. Stage MT, the number of stage shifts, and integral EMG increased significantly from the fifth night and later, showing p less than 0.01, p less than 0.01, and p less than 0.05, respectively. Judging from these findings, the sleep habituation of the subject in the laboratory was completed within the first four nights. Coefficients of variation of sleep stage 2 and stage REM of the ten-nights' EEG were the lowest among all the sleep parameters examined. Almost all the parameters of day-to-day sleep of the subject who slept for ten successive nights in the laboratory showed smaller variations than those of the other six subjects. It may be concluded that the mist effect on sleep could be assessed more precisely by using an individual repeatedly than by using a group of subjects.

Adult↗

Airborne concentrations of PM(2.5) and diesel exhaust particles on Harlem sidewalks: a community-based pilot study.

Residents of the dense urban core neighborhoods of New York City (NYC) have expressed increasing concern about the potential human health impacts of diesel vehicle emissions. We measured concentrations of particulate matter [less than/equal to] 2.5 micro in aerodynamic diameter (PM(2.5)) and diesel exhaust particles (DEP) on sidewalks in Harlem, NYC, and tested whether spatial variations in concentrations were related to local diesel traffic density. Eight-hour (1000-1800 hr) air samples for PM(2.5 )and elemental carbon (EC) were collected for 5 days in July 1996 on sidewalks adjacent to four geographically distinct Harlem intersections. Samples were taken using portable monitors worn by study staff. Simultaneous traffic counts for diesel trucks, buses, cars, and pedestrians were carried out at each intersection on [Greater/equal to] 2 of the 5 sampling days. Eight-hour diesel vehicle counts ranged from 61 to 2,467 across the four sites. Mean concentrations of PM(2.5) exhibited only modest site-to-site variation (37-47 microg/m(3)), reflecting the importance of broader regional sources of PM(2.5). In contrast, EC concentrations varied 4-fold across sites (from 1.5 to 6 microg/m(3)), and were associated with bus and truck counts on adjacent streets and, at one site, with the presence of a bus depot. A high correlation (r = 0.95) was observed between EC concentrations measured analytically and a blackness measurement based on PM(2.5) filter reflectance, suggesting the utility of the latter as a surrogate measure of DEP in future community-based studies. These results show that local diesel sources in Harlem create spatial variations in sidewalk concentrations of DEP. The study also demonstrates the feasibility of a new paradigm for community-based research involving full and active partnership between academic scientists and community-based organizations.

Academic Medical Centers↗

Validation and calibration of a model used to reconstruct historical exposure to polycyclic aromatic hydrocarbons for use in epidemiologic studies.

OBJECTIVES: We previously developed a historical reconstruction model to estimate exposure to airborne polycyclic aromatic hydrocarbons (PAHs) from traffic back to 1960 for use in case-control studies of breast cancer risk. Here we report the results of four exercises to validate and calibrate the model. METHODS: Model predictions of benzo[a]pyrene (BaP) concentration in soil and carpet dust were tested against measurements collected at subjects' homes at interview. In addition, predictions of air intake of BaP were compared with blood PAH-DNA adducts. These same soil, carpet, and blood measurements were used for model optimization. In a separate test of the meteorological dispersion part of the model, predictions of hourly concentrations of carbon monoxide from traffic were compared with data collected at a U.S. Environmental Protection Agency monitoring station. RESULTS: The data for soil, PAH-DNA adducts, and carbon monoxide concentrations were all consistent with model predictions. The carpet dust data were inconsistent, suggesting possible spatial confounding with PAH-containing contamination tracked in from outdoors or unmodeled cooking sources. BaP was found proportional to other PAHs in our soil and dust data, making it reasonable to use BaP historical data as a surrogate for other PAHs. Road intersections contributed 40-80% of both total emissions and average exposures, suggesting that the repertoire of simple markers of exposure, such as traffic counts and/or distance to nearest road, needs to be expanded to include distance to nearest intersection.

Calibration↗

"The very house of difference": Zami, Audre Lorde's lesbian-centered text.

Locating the feminist subject at the intersection of shifting social groups, or "homes," allows me to explore the connections between concepts of home and identity in Audre Lorde's "biomythography," Zami: A New Spelling of My Name. While Lorde's writing attends to the complex intersections of race, gender, and sexuality, as the prologue to the text makes apparent, it also testifies to the "naturalness" of lesbianism as the ground of writing, thinking, and acting. In Lorde's text, the lesbian body figures neither as an essential or fixed identity nor as the site for a unified conception of community or home, but rather, as a paradigm for a new kind of writing--one which inhabits the very house of difference.

Black or African American↗

Ambiguity and the image of the king.

The following essay explores problems posed by a recently-published fresco (dated to the first century AD) that depicts Alexander the Great standing opposite an unknown female figure. The fresco is unusual in its use of conventional or codified figure types, in particular a widely-found statue type known as the "Alexander with the Lance," and for its placement of Alexander in anecdotal relation with a woman. While discussions of the picture thus far have tried to identify the scene depicted (by reference to histories of Alexander's life), the following analysis takes the difficulty of doing so itself as a motivated aspect of the image. I argue that the fresco's mode of representation is to bring together figure types whose conventional fields of meaning are in conflict with one another, and then to highlight this conflict in order to comment upon the fields (or figure types) themselves. In this case, the fresco's ambiguity in signification (the undecidability of its reference) enables a highly strategic critique of the "Alexander with the Lance" because the latter, as a prototypical "image of the king," depends upon the necessary and transparent extension of its signs. By virtue of the anecdotal relation between "Alexander" and the depicted female figure (an Aphrodite type) the fresco's critique reveals the close association between the claims for representation made by the image of the king and the patriarchal structures of power they seek to instantiate. The fresco thus offers remarkably direct data for understanding the intersection of representation and gender in the early Roman empire. I suggest in conclusion that because the image seems also to posit a specifically gendered (male) gaze, its critique is extended to the spectator and thereby provides data for understanding the intersection of the practice of representation (here, viewing) and gender.

Gender Identity↗

Method-based differences in the automated analysis of the three-dimensional morphology of trabecular bone.

The three-dimensional (3D) morphology of trabecular bone is frequently quantified using computer programs. However, there are no standardized implementations of morphology programs and many variations are possible. Even though programs may use the same basic method, results can be significantly different because of differences in implementation. Morphology data from different laboratories therefore may not be comparable. The method of directed secants, with parallel plate assumptions, is commonly used to quantify 3D morphology. We examined the effect of several variations in the implementation of this method on measurements of trabecular plate number (Tb.N), trabecular thickness, and trabecular spacing. Three-dimensional micromagnetic resonance images of 10 bovine trabecular bone specimens were analyzed using several variations of the directed secant method. An analysis of covariance with repeated measures suggested that variations in the algorithm used to count test line intersections, variations in the criteria used to classify a test coordinate as bone or marrow, and variations in the number of test grid rotations had significant effects on Tb.N (p < 0.0001). The largest difference in Tb.N (52%) was due to the method used to count test line intersections with the bone-marrow interface. Variations in the classification algorithm and variations in the number of test line grid rotations resulted in a 6% difference in Tb.N. The spacing of the test line grids did not affect Tb.N (p = 0.28), and all differences were independent of volume fraction (p = 0.67). These data show that there can be significant differences in trabecular bone morphology measurements due only to the method used for the measurements. To facilitate comparisons between laboratories, we have made validated computer programs to measure trabecular bone morphology available over the Internet.

Algorithms↗

Microinstrument gradient-force optical trap.

A micromachined fiber-optic trap is presented. The trap consists of four single-mode, 1064-nm optical intersection. The beam fibers mounted in a micromachined silicon and glass housing. Micromachining provides the necessary precision to align the four optical fibers so that the outputs have a common intersection forms a strong three-dimensional gradient-force trap with trapping forces comparable with that of optical tweezers. Characterization of the multibeam fiber trap is illustrated for capture of polystyrene microspheres, computer simulations of the trap stiffness, and experimental determination of the trapping forces.

Computer Simulation↗

Comparison of measurements made with two different instruments of the same atmospheric vertical profile.

The validation of atmospheric remote-sensing measurements involves the comparison of vertical profiles of atmospheric constituents obtained by different instruments. This operation is a complex one because it has to take into account the measurement errors that are described by the variance-covariance matrices and the different features of the two observing systems that are described by the averaging kernels. The procedure is discussed and a method of comparison that is rigorous and does not involve degradation of the available information is developed by use of the formalism of functional spaces. The functional spaces that can be used for representation of the two profiles are reviewed, and criteria are determined for the choice of the most convenient functional space to minimize degradation of the measurements. Once the functional spaces are chosen, the components of the profiles are compared in the intersection space of these two functional spaces. If the intersection space coincides with the null vector, a pseudointersection space with useful geometrical properties can be used instead. A test of the method is made with a realistic simulation. In the test the profiles retrieved by two real instruments are simulated and quantitatively compared.

Journal Article↗

The importance of outcomes research in pediatric emergency medicine.

Applying the methods and tools of outcomes research, "evaluation of the impact of health care on the health outcomes or 'end results' of patients and populations," to the clinical domain of emergency services for children offers an important strategic opportunity for addressing the questions that confront all health care services: What works? For which patients? At what cost? From whose perspective? Although the important questions to address are extensive, much of the intersection between emergency services and outcomes research remains unexplored. Important challenges for researchers intrigued by the opportunities at this intersection of fields include: 1) clear definition of the scope of emergency services; 2) consideration of the appropriate end-point of emergency services-the entire episode of illness and/or services provided within the emergency setting; 3) selection and development of measures that incorporate children's and families' perspectives; and 4) a clear focus on linking research findings with strategies for improving outcomes and informed decision making. This essay will provide an overview of accomplishments and challenges from the field of outcomes research, suggest important opportunities for applying existing methods to emergency medical services for children, and identify potential career paths for current and future investigators.

Child↗

Screening of core targets for Di(2-ethylhexyl) Phthalate-related gastric cancer based on machine learning, molecular docking, and SHAP analysis.

PURPOSE: Given the existing uncertainties regarding the link between Di(2-ethylhexyl) phthalate (DEHP) exposure and gastric cancer (GC) progression, this study aimed to clarify their association, identify the toxic targets of DEHP, and elucidate the underlying molecular mechanisms. METHODS: Multiple integrated approaches were employed, including Gene Expression Omnibus (GEO) data analysis, network toxicology, molecular docking, and machine learning. STRING and Cytoscape tools were utilized to identify key targets, while Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to explore the functional enrichment of intersecting targets. Machine learning and SHAP analysis were applied to screen core targets in GC. Molecular docking was performed to evaluate the binding affinity of DEHP toward core targets, and 200 ns molecular dynamics simulations were further conducted for representative complexes to validate their dynamic stability. RESULTS: A total of 18 key targets were identified using STRING and Cytoscape. GO and KEGG enrichment analyses demonstrated that these intersecting targets were primarily enriched in the extracellular region, as well as the Calcium signaling pathway and cAMP signaling pathway. Through machine learning analyses, 7 key genes (ADRB2, ESRRG, GRIA4, IL13RA2, NR3C2, PLA2G1B, and SULT2A1) were identified as core targets in GC through machine learning analyses. Molecular docking simulations revealed strong binding specificity between DEHP and the target proteins. Among them, NR3C2 and ADRB2 exhibited relatively high predictive importance in the machine learning models. DEHP showed favorable binding affinity toward these core targets, and molecular dynamics simulations further confirmed that ADRB2-DEHP and NR3C2-DEHP complexes maintained stable conformations throughout the simulation. CONCLUSIONS: Our findings identified GC associated genes that were computationally predicted as potential targets of DEHP. These results indicated structural compatibility between DEHP and its target proteins but did not prove that DEHP exposure accounts for the gene expression changes in GC.

Molecular Docking Simulation↗

The proportional Venn diagram of obstructive lung disease: two approximations from the United States and the United Kingdom.

STUDY OBJECTIVES: The nonproportional Venn diagram of obstructive lung disease (OLD) produced for the 1995 American Thoracic Society guidelines has not been quantified. We aim to quantify the proportion of the general population with OLD and the intersections of physician-diagnosed asthma, chronic bronchitis, and emphysema in the United States and the United Kingdom, and to examine the relationship to obstructive spirometry. DESIGN AND PARTICIPANTS: We analyzed data from the US National Health and Nutrition Examination (NHANES) III survey (1988 to 1994) and the UK General Practice Research Database for the year 1998. RESULTS: The areas of intersection among the three OLD conditions produced seven mutually exclusive disease groups. The asthma-only group was the largest proportion of OLD patients, accounting for 50.3% and 79.4% of all OLD patients in the United States and the United Kingdom, respectively, and decreased with increasing age. Overall, 17% and 19% of OLD patients in the United States and in the United Kingdom, respectively, reported more than one OLD condition, and this percentage increased with age. According to the spirometry data from NHANES III, only 37.4% of emphysema-only patients had objective airflow obstruction. The prevalence of airflow obstruction was significantly higher among participants with combinations of emphysema and chronic bronchitis (57.7%), with emphysema and asthma (51.9%), and with all three OLD diseases concomitantly (52.0%). CONCLUSION: Concomitant diagnosis of asthma, chronic bronchitis, or emphysema is common among OLD patients from the general population, particularly in adults aged > or = 50 years.

Adult↗

The triadic intersubjective matrix in supervision: the use of disclosure to work through painful affects.

The use of the psychoanalyst's subjective reactions as a tool to better understand his/her patient has been a central feature of clinical thinking in recent decades. While there has been much discussion and debate about the analyst's use of countertransference in individual psychoanalysis, including possible disclosure of his/her feelings to the patient, the literature on supervision has been slower to consider such matters. The attention to parallel processes in supervision has been helpful in appreciating the impact of affects arising in either the analyst/patient or the supervisor/analyst dyads upon the analytic treatment and its supervision. This contribution addresses the ways in which overlapping aspects of the personalities of the supervisor, analyst and patient may intersect and create resistances in the treatment. That three-way intersection, described here as the triadic intersubjective matrix, is considered inevitable in all supervised treatments. A clinical example from the termination phase of a supervised analysis of an adolescent is offered to illustrate these points. Finally, the question of self-disclosure as an aspect of the supervisory alliance is also discussed.

Adolescent↗