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Osteoporosis is defined as a skeletal disorder characterized by compromised bone strength. Bone strength depends, among others, on bone density, bone geometry and its internal architecture. With the recent introduction of a new generation high-resolution 3D peripheral quantitative computed tomography (HR-pQCT) system, direct quantification of structural bone parameters has become feasible. Furthermore, it has recently been demonstrated that bone mechanical competence can be derived from HR-pQCT based micro-finite element modeling (microFE). However, reproducibility data for HR-pQCT-derived mechanical indices is not well-known. Therefore, the aim of this study was to quantify reproducibility of HR-pQCT-derived indices. We measured 14 distal formalin-fixed cadaveric forearms three times and analyzed three different regions for each measurement. For each region cortical and trabecular parameters were determined. Reproducibility was assessed with respect to precision error (PE) and intraclass correlation coefficient (ICC). Reproducibility values were found to be best in all three regions for the full bone compartment with an average PE of 0.79%, followed by the cortical compartment (PE=1.19%) and the trabecular compartment with an average PE of 2.31%. The mechanical parameters showed similar reproducibility (PE=0.48%-2.93% for bone strength and stiffness, respectively). ICC showed a very high reproducibility of subject-specific measurements, ranging from 0.982 to 1.000, allowing secure identification of individual donors ranging from healthy to severely osteoporotic subjects. From these in vitro results we conclude that HR-pQCT derived morphometric and mechanical parameters are highly reproducible such that differences in bone structure and strength can be detected with a reproducibility error smaller than 3%; hence, the technique has a high potential to become a tool for detecting bone quality and bone competence of individual subjects.

The homeobox gene bicoid functions as an anterior pattern organizer of the Drosophila embryo, but other than in higher flies (Cyclorrhapha), bicoid orthologues appear to be absent from insect genomes. In Drosophila, bicoid is expressed in an anterior-to-posterior protein gradient and regulates spatially restricted expression domains of segmentation genes in a concentration-dependent manner. hunchback, a direct transcriptional target of Bicoid, complements the "morphogen" activity of Bicoid. hunchback is activated by Bicoid throughout the anterior half of the blastoderm and a Bicoid-binding cis-regulatory element has been identified immediately upstream of the proximal hunchback promoter P2 of Drosophila and other higher Cyclorrhapha (Schizophora). bicoid and Bicoid-dependent hunchback regulation are thought to have originated during or before the radiation of extant Cyclorrhapha, although the precise occurrence of these traits in lower Cyclorrhapha remains unknown. Previously, we have described a bicoid orthologue in Megaselia, a species of the lower cyclorrhaphan family Phoridae. Here, we report the occurrence of bicoid in two additional lower cyclorrhaphan families, Lonchopteridae and Platypezidae. We show that Megaselia Bicoid is required for anterior expression of Megaselia hunchback, and binds upstream of its P2 promoter. Furthermore, we report the expression of lacZ reporter constructs under the control of hunchback regulatory sequences from a range of lower cyclorrhaphan and non-cyclorrhaphan flies in transgenic Drosophila embryos. Our results are consistent with a cyclorrhaphan origin of bicoid and suggest that a Bicoid-binding enhancer upstream of the hunchback P2 promoter evolved at the latest in the last common ancestor of Megaselia and Schizophora.

The homeobox gene caudal (cad) regulates posterior development in Drosophila. In early embryos, the cad protein (CAD) is expressed in a posterior-to-anterior concentration gradient, which contributes polarity to the developing embryo. The CAD gradient is complementary to and dependent on the anterior pattern organizer Bicoid (BCD), which represses the translation of ubiquitous maternal cad transcripts in the anterior embryo through a direct interaction with the cad 3' untranslated region (UTR). Here, we show that early embryos of the lower cyclorrhaphan fly Megaselia express the putative cad orthologue Mab-cad throughout the posterior three quarters of the blastoderm but lack maternal transcripts. In transgenic blastoderm embryos of Drosophila, Mab-cad cis-regulatory DNA drives the expression of a reporter gene in a similar pattern, while Mab-cad 3' UTR fails to mediate translational repression of a ubiquitously transcribed reporter. For another lower cyclorrhaphan fly (Lonchoptera) and two related outgroup taxa of Cyclorrhapha (Empis, Haematopota), we report maternal cad expression in ovarian follicles. Together, our results suggest that BCD is not required for the translational repression of Mab-cad, and that maternal cad expression was lost in the Megaselia lineage.

Higher cyclorrhaphan flies including Drosophila develop a single extraembryonic epithelium (amnioserosa), which closes the germband dorsally. In most other insects two extraembryonic epithelia, serosa and amnion, line the inner eggshell and the ventral germband, respectively. How the two extraembryonic epithelia evolved into one is unclear. Recent studies have shown that, in the flour beetle Tribolium and in the milkweed bug Oncopeltus, the homeobox gene zerknüllt (zen) controls the fusion of the amnion with the serosa before dorsal closure. To understand the origin of the amnioserosa in evolution, we examined the expression and function of zen in the extraembryonic tissue of lower Cyclorrhapha. We show that Megaselia abdita (Phoridae) and Episyrphus balteatus (Syrphidae) develop a serosa and a dorsal amnion, suggesting that a dorsal amnion preceded the origin of the amnioserosa in evolution. Using Krüppel (Kr) and pannier (pnr) homologues of Megaselia as markers for serosal and amniotic tissue, respectively, we show that after zen RNAi all extraembryonic tissue becomes indistinguishable from amniotic cells, like in Tribolium but unlike in Drosophila, in which zen controls all aspects of extraembryonic development. Compared with Megaselia and Episyrphus, zen expression in Drosophila is extended to cells that form the amnion in lower Cyclorrhapha and is down-regulated at the developmental stage, when serosa cells in lower Cyclorrhapha begin to expand. These expression differences between species with distinct extraembryonic tissue organizations and the conserved requirement of zen for serosa development suggest that the origin of an amnioserosa-like epithelium was accompanied by expression changes of zen.

The large increase in interest in micro-computed tomography (micro-CT) over the last decade reflects the+ need for a method able to non-destructively visualize the internal three-dimensional structure of an object. Thereby, the real beauty of computed tomography lies in the fact that it is available for a large range of nominal resolutions, which allows hierarchical imaging from whole bodies down to the tissue level. Although micro-CT is currently mainly used for imaging of hard tissue (i.e., bone and tooth), future developments might also allow high soft tissue contrast either using appropriate contrast agents or x-ray contrast mechanisms. This chapter aims to review the steps necessary for a successful micro-CT measurement. Although the actual measurement is often machine dependent, the chapter does not describe a specific system but rather lists all steps that eventually have to be considered to set up a measurement, run the measurement, process the image data, and get morphometric indices as a result. The chapter provides an easy understandable manual that should allow newcomers to perform successful measurements and hence to best profit from this powerful technique.

Unlabelled: Nondestructive SR-based microCT and nano-CT methods have been designed for 3D quantification and morphometric analysis of ultrastructural phenotypes within murine cortical bone, namely the canal network and the osteocyte lacunar system. Results in two different mouse strains, C57BL/6J-Ghrhr(lit)/J and C3.B6-Ghrhr(lit)/J, showed that the cannular and lacunar morphometry and their bone mechanics were fundamentally different., Introduction: To describe the different aspects of bone quality, we followed a hierarchical approach and assessed bone tissue properties in different regimens of spatial resolution, beginning at the organ level and going down to cellular dimensions. For these purposes, we developed different synchrotron radiation (SR)-based CT methods to assess ultrastructural phenotypes of murine bone., Materials and Methods: The femoral mid-diaphyses of 12 C57BL/6J-Ghrhr(lit)/J (B6-lit/lit) and 12 homozygous mutants C3.B6-Ghrhr(lit)/J (C3.B6-lit/lit) were measured with global SR microCT and local SR nano-CT (nCT) at nominal resolutions ranging from 3.5 microm to 700 nm, respectively. For volumetric quantification, morphometric indices were determined for the cortical bone, the canal network, and the osteocyte lacunar system using negative imaging. Moreover, the biomechanics of B6-lit/lit and C3.B6-lit/lit mice was determined by three-point bending., Results: The femoral mid-diaphysis of C3.B6-lit/lit was larger compared with B6-lit/lit mice. On an ultrastructural level, the cannular indices for C3.B6-lit/lit were generally bigger in comparison with B6-lit/lit mice. Accordingly, we derived and showed a scaling rule, saying that overall cannular indices scaled with bone size, whereas indices describing basic elements of cannular and lacunar morphometry did not. Although in C3.B6-lit/lit, the mean canal volume was larger than in B6-lit/lit, canal number density was proportionally smaller in C3.B6-lit/lit, so that lacuna volume density was found to be constant and therefore independent of mouse strain and sex. The mechanical properties in C3.B6-lit/lit were generally improved compared with B6-lit/lit specimens. For C3.B6-lit/lit, we observed a sex specificity of the mechanical parameters, which could not be explained by bone morphometry on an organ level. However, there is evidence that for C3.B6-lit/lit, the larger cortical bone mass is counterbalanced or even outweighed by the larger canal network in the female mice., Conclusions: We established a strategy to subdivide murine intracortical porosity into ultrastructural phenotypes, namely the canal network and the osteocyte lacunar system. Nondestructive global and local SR-based CT methods have been designed for 3D quantification and subsequent morphometric analysis of these phenotypes. Results in the two different mouse strains C57BL/6J-Ghrhr(lit)/J and C3.B6-Ghrhr(lit)/J showed that the cannular and lacunar morphometry and the biomechanical properties were fundamentally different.

Mouse models are widely used in genomic studies for quantitative trait loci (QTL) analyses. In the field of skeletal micro-structure, microCT has proven to be an invaluable imaging tool for the characterization of structural bone traits. However, the definition of analysis compartments requires a lot of user interaction, and therefore is not applicable as a standard way to analyze genetic linkage studies with several hundreds of animals. Here, we developed an automated three-dimensional based algorithm for a high-throughput regional analysis of three compartments in murine femora, including whole bone, cortical bone in the diaphysis and trabecular bone in the metaphysis. The algorithm relies on basic image processing concepts using morphological operators as well as a new approach of separating cortical from trabecular bone. Reproducibility of the automatic approach was investigated with respect to precision errors (PE(%CV)) of micro-structural indices analyzed in these automatically defined compartments. The developed algorithm was then used to perform a high-throughput analysis of over 2000 femora in a genetic linkage study for further examination of stability and performance. Precision errors were 3.5% or less for all micro-structural indices. The analysis of one femur (mask generation and parameter evaluation) took 7 min on an AlphaServer DS25. The algorithm showed a very high reliability and worked successfully for 99.64% of all femora. Investigations of correlations amongst the assessed micro-structural indices together with heritability and polygene estimates revealed apparent volume density (AVD), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp) and cortical thickness (Ct.Th) as candidates for a successful QTL analysis. The presented automatic analysis allows for standardized high-throughput phenotypic screening in mice femora for large genetic linkage studies with very high reliability and good precision.

Recently, we proposed a new computational approach for the volumetric spatial decomposition of a three-dimensional bone structure into its basic rod and plate elements. This method was based on an image skeletonization approach, where two model parameters were used to identify an ideal skeleton. The goal of this study was to estimate the sensitivity of local morphometric indices to these two model parameters. Our results showed that the rod derived indices behaved more smoothly than plate derived indices, which suggests that rod derived indices are more trustworthy. The results also demonstrated that it was reasonable to reduce the model to only one parameter by setting the noise parameter n to twice the value of the slenderness parameter s. In conclusion, we found that local morphometric indices are reliable measures showing large differences between samples and thus may shed new light on structural differences of trabecular bone in a local fashion by adequately choosing one single optimization parameter.

Inroads have been made in the diagnosis and treatment of osteoporosis, yet dual-energy X-ray absorptiometry is still the primary diagnostic modality. This method provides 2D projections of an irregular 3D construct. However, human cancellous bone is highly heterogeneous with varying material properties. Therefore, to properly assess fracture risk, it is imperative to take into consideration microstructural indices besides subregional bone volume fraction (BV/TV). A power law model with average BV/TV as the independent variable describes 38% of the variation in yield strength; however, this predictive power is increased to 56% when BV/TV of the weakest subregion is considered. Of twenty-five specimens studied, 76% had minimum BV/TV, maximum principal Eigen value of the fabric tensor (H1) and minimum connectivity density (Conn.D) values within the visually determined failure regions. These three independent morphometric indices yielded significant differences between the failure and non-failure regions of each specimen. From the results, we conclude that subregions with minimal BV/TV values are better predictors of mechanical failure in cancellous bone than average specimen BV/TV. Addition of microstructural indices augments this predictive power to generate a trabecular failure prediction model based on volume fraction and cancellous bone microstructure specifically in areas where trabecular failure is most likely to occur.

Direct assessment of bone competence in vivo is not possible, hence, it is inevitable to predict it using appropriate simulation techniques. Although accurate estimates of bone competence can be obtained from micro-finite element models (muFE), it is at the expense of large computer efforts. In this study, we investigated the application of structural idealizations to represent individual trabeculae by single elements. The objective was to implement and validate this technique. We scanned 42 human vertebral bone samples (10 mm height, 8 mm diameter) with micro-computed tomography using a 20 microm resolution. After scanning, direct mechanical testing was performed. Topological classification and dilation-based algorithms were used to identify individual rods and plates. Two FE models were created for each specimen. In the first one, each rod-like trabecula was modeled with one thickness-matched beam; each plate-like trabecula was modeled with several beams. From a simulated compression test, assuming one isotropic tissue modulus for all elements, the apparent stiffness was calculated. After reducing the voxel size to 40 microm, a second FE model was created using a standard voxel conversion technique. Again, one tissue modulus was assumed for all elements in all models, and a compression test was simulated. Bone volume fraction ranged from 3.7% to 19.5%; Young's moduli from 43 MPa to 649 MPa. Both models predicted measured apparent moduli equally well (R2 = 0.85), and were in excellent agreement with each other (R2 = 0.97). Tissue modulus was estimated at 9.0 GPa and 10.7 GPa for the beam FE and voxel FE models, respectively. On average, the beam models were solved in 219 s, reducing CPU usage up to 1150-fold as compared to 40 microm voxel FE models. Relative to 20 microm voxel models 10,000-fold reductions can be expected. The presented beam FE model is an abstraction of the intricate real trabecular structure using simple cylindrical beam elements. Nevertheless, it enabled an accurate prediction of global mechanical properties of microstructural bone. The strong reduction in CPU time provides the means to increase throughput, to analyze multiple loading configuration and to increase sample size, without increasing computational costs. With upcoming in vivo high-resolution imaging systems, this model has the potential to become a standard for mechanical characterization of bone.

Synchrotron radiation micro-computed tomography (SRmicroCT) is a very useful technique when it comes to three-dimensional (3D) imaging of complex internal and external geometries. Being a fully non-destructive technique, SRmicroCT can be combined with other experiments in situ for functional imaging. We are especially interested in the combination of SRmicroCT with mechanical testing in order to gain new insights in the failure mechanism of trabecular bone. This interest is motivated by the immense costs in health care due to patients suffering from osteoporosis, a systemic skeletal disease resulting in decreased bone stability and increased fracture risk. To better investigate the different failure mechanisms on the microlevel, we have developed a novel in situ mechanical compression device, capable of exerting both static and dynamic displacements on experimental samples. The device was calibrated for mechanical testing using solid aluminum and bovine trabecular bone samples. To study different failure mechanisms in trabecular bone, we compared a fatigued and a non-fatigued bovine bone sample with respect to failure initiation and propagation. The fatigued sample failed in a burst-like fashion in contrast to the non-fatigued sample, which exhibited a distinct localized failure band. Moreover, microscopic cracks - microcracks and microfractures - were uncovered in a 3D fashion illustrating the failure process in great detail. The majority of these cracks were connected to a bone surface. The data also showed that the classification of microcracks and -fractures from 2D section can sometimes be ambiguous, which is also true for the distinction of diffuse and distinct microdamage. Detailed investigation of the failure mechanism in these samples illustrated that trabecular bone often fails in delamination, providing a mechanism for energy dissipation while conserving trabecular bone architecture. In the future, this will allow an even better understanding of bone mechanics related to its hierarchical structural organization.

The goal of our study was to evaluate two newly developed implant designs and their behavior in terms of subsidence in lumbar vertebral bodies under cyclic loading. The new implants were evaluated in two different configurations (two small prototypes vs. one large prototype with similar load-bearing area) in comparison to a conventional screw-based implant (MACS TL). A pool of 13 spines with a total of 65 vertebrae was used to establish five testing groups of similar bone mineral density (BMD) distribution with eight lumbar vertebrae each. In additional to BMD assessment via dual-energy X-ray absorptiometry, cancellous BMD and structural parameters were determined using a new generation in vivo 3D-pQCT. The specimens were loaded sinusoidally in force control at 1 Hz for 1000 cycles at three load levels (100, 200, and 400 N). A survival analysis using the number of cycles until failure (Cox regression with covariates) was applied to reveal differences between implant groups. All new prototype configurations except the large cylinder survived significantly longer than the control group. The number of cycles until failure was significantly correlated with the structural parameter Tb.Sp. and similarly with the cancellous BMD for three of five implants. In both large prototypes the cycle number until failure significantly correlated with the preoperative distance to the upper endplates. Although the direct relationship between bone structure or density and mechanical breakage behavior cannot be conclusively proven, all the prototypes adapted for poor bone structure performed better than the comparable conventional implant., (Copyright 2006 Orthopaedic Research Society.)

Unlabelled: Local morphometry based on the assessment of individual rods and plates was applied to 42 human vertebral trabecular bone samples. Results showed that multiple linear regression models based on local morphometry as a measure for bone microstructure helped improving our understanding of the role of local structural changes in the determination of bone stiffness as assessed from direct and computational biomechanics., Introduction: In a recent study, we proposed a method for local morphometry of trabecular bone, i.e., morphometry as applied to individual rods and plates. In this study, we used this method to study the relative importance of local morphometry in the assessment of bone architecture and its relative contribution to the stiffness of human vertebral bone., Materials and Methods: We extracted 42 human trabecular bone autopsies from nine intact spinal columns. The cylindrical samples were imaged with muCT to assess bone microstructure. From these images, global and local morphometric indices were derived and related to Young's modulus as assessed by experimental uniaxial compression testing (Emeas) and computational finite element analysis (EFE)., Results: We found the best single predictor for Young's modulus to be apparent bone volume density (BV/TV), which explained 89% of the variance in EFE when fitted with a power law. A multiple linear regression model combining mean trabecular spacing (Tb.Sp), mean slenderness of the rods (), and the relative amount of rod volume to total bone volume (Ro.BV/BV) was able to explain 90% of the variance in EFE. This model could not be improved by adding BV/TV as an independent variable. Furthermore, we found that mean trabecular thickness of the rods was significantly related to EFE (r2 = 0.42), whereas mean trabecular thickness of plates had no correlation to Young's modulus. Because the globally determined trabecular thickness does not discriminate between rods and plates, this index had only a poor predictive power for EFE (r2 = 0.09), showing the importance of local analysis of individual rods and plates., Conclusions: From these results, we conclude that models based on local morphometry help improving our understanding of the relative importance of local structural changes in the determination of the stiffness of bone. Separate analysis of individual rods and plates may help to better predict age and disease-related fractures as well as to shed new light on the effect of pharmaceutical intervention in the prevention of such fractures beyond BMD.

Bone microarchitecture is believed to play a key role in determining bone quality. We therefore present a new method for the volumetric spatial decomposition of trabecular bone samples into its basic elements (rods and plates). This new method is a framework for the element based description of bone microarchitecture. First, the newly developed algorithm was validated on computer-generated models. Then, it was applied to 328 human trabecular bone samples harvested from 70 donors at five different anatomical sites (calcaneus, femoral head, iliac crest, lumbar spine 2 and 4), which were previously scanned by microcomputed tomography. Standard three-dimensional morphometric algorithms were used to analyze the trabeculae on an individual basis with respect to their volume, surface, and thickness. The results were statistically compared for the five sites. In this study, it was possible for the first time to spatially decompose trabecular bone structures in its volumetric elements; rods and plates. The size of the largest element in the structures showed significant differences for the five compared sites. In samples from femoral head, we found that basically one "major element" was spanning through the whole structure whereas in lumbar spine and calcaneus, smaller elements dominate. From this, we suggest that the strength of strong, dense plate-like structures is determined by the major elements whereas in looser rod-like structures the strength is given by the arrangement, quality, and shape of a whole set of elements. Furthermore, we found that globally determined structural indices such as the mean curvature of the bone surface () or related to this the structure model index (SMI) are almost exclusively explained by the arrangement of the plates. This also suggests that rods hold independent information characterizing trabecular bone quality, especially in the spine. These findings may improve the understanding of the site-specific role of bone microarchitecture in determining bone quality and in future studies the competence of bone.

Introduction: A recently developed method allows investigating trabecular bone on an elemental (rod/plate) level. With this method, it is possible to measure local morphometric parameters such as thickness or orientation directly on the extracted rods and plates. Age-related changes of trabecular microarchitecture can thus be investigated on an elemental level, which may help to improve the understanding of age-related bone failure mechanism as well as the effect of pharmaceutical intervention in the prevention of such fractures., Methods: Autopsies from femoral heads (FH) and lumbar spine (LS) were analyzed by global morphometry. Additionally, the trabecular structures were decomposed into rods and plates for the analysis with local morphometry. These morphometric indices were related to age using an analysis of covariance to test for gender differences and linearity with age., Results: In this study, age-related changes showed no gender but site differences. In LS, rods were thinned in aging and finally vanish from the structure, causing a transformation of the trabecular bone structure to longer and, on average, thicker rods. In FH, changes were expressed by a simultaneous thinning and loss of interconnecting trabeculae and perforation of plates leading to new plates and rods. Results were mostly in agreement with earlier findings using descriptive analysis of the aging process., Conclusion: Here we present for the first time preliminary quantitative evidence of changes in the local microstructure, i.e., individual rods and plates. Nevertheless, the number of samples was too small to make for ready conclusions. We conclude that the combination of local and global morphometry is a useful method for a detailed and quantitative description of age-related changes in bone microstructure.

A collaborative study on Raman spectroscopy was carried out by members of the ENFSI (European Network of Forensic Science Institutes) European Fibres Group (EFG) on three dyed fibers: two red acrylics and one red wool. Raman instruments from six different manufacturers were tested as well as nine different laser wavelengths ranging from blue (lambda = 458 nm) to near infrared-NIR (lambda = 1064 nm). This represents the largest comparison study of Raman analytical parameters carried out on identical fiber samples. For the chosen fiber and dye samples, red lasers (lambda = 633 and 685 nm) gave the poorest spectral quality whereas blue (458 nm), green (514 nm) and near infrared lasers (785, 830 and 1064 nm) provided average results. Blue (488 nm) and green lasers (532 nm) globally gave the best quality spectra. Fluorescence problems were often encountered with some of the excitation wavelengths and therefore a flexible Raman instrument equipped with different lasers can be recommended to measure forensic fiber samples. The instrument should also be equipped with a Raman microscope in order to be able to focus on a single fiber. This study shows that Raman spectroscopy usually enables the identification of the main dye present in a colored fiber; however, minor dye components are much more difficult to detect. SERRS (Surface Enhanced Resonance Raman Scattering) techniques give an improvement of the dye's spectral intensity but no spectral improvement was observed for the two red acrylic and red wool fibers tested.

Cellular solids constitute an important class of engineering materials encompassing both man-made and natural constructs. Materials such as wood, cork, coral, and cancellous bone are examples of cellular solids. The structural analysis of cellular solid failure has been limited to 2D sections to illustrate global fracture patterns. Due to the inherent destructiveness of 2D methods, dynamic assessment of fracture progression has not been possible. Image-guided failure assessment (IGFA), a noninvasive technique to analyze 3D progressive bone failure, has been developed utilizing stepwise microcompression in combination with time-lapsed microcomputed tomographic imaging (microCT). This method allows for the assessment of fracture progression in the plastic region, where much of the structural deformation/energy absorption is encountered in a cellular solid. Therefore, the goal of this project was to design and fabricate a novel micromechanical testing system to validate the effectiveness of the stepwise IGFA technique compared to classical continuous mechanical testing, using a variety of engineered and natural cellular solids. In our analysis, we found stepwise compression to be a valid approach for IGFA with high precision and accuracy comparable to classical continuous testing. Therefore, this approach complements the conventional mechanical testing methods by providing visual insight into the failure propagation mechanisms of cellular solids., ((c) 2005 Wiley Periodicals, Inc.)

Establishment of segmental pattern in the Drosophila syncytial blastoderm embryo depends on pair-rule transcriptional regulators. mRNA transcripts of pair-rule genes localise to the apical cytoplasm of the blastoderm via a selective dynein-based transport system and signals within their 3'-untranslated regions. However, the functional and evolutionary significance of this process remains unknown. We have analysed subcellular localisation of mRNAs from multiple dipteran species both in situ and by injection into Drosophila embryos. We find that although localisation of wingless transcripts is conserved in Diptera, localisation of even-skipped and hairy pair-rule transcripts is evolutionarily labile and correlates with taxon-specific changes in positioning of nuclei. We show in Drosophila that localised pair-rule transcripts target their proteins in close proximity to the nuclei and increase the reliability of the segmentation process by augmenting gene activity. Our data suggest that mRNA localisation signals in pair-rule transcripts affect nuclear protein uptake and thereby adjust gene activity to a variety of dipteran blastoderm cytoarchitectures.

This paper presents a new method for the generation of a beam finite element (FE) model from a three-dimensional (3D) data set acquired by micro-computed tomography (micro-CT). This method differs from classical modeling of trabecular bone because it models a specific sample only and differs from conventional solid hexahedron element-based FE approaches in its computational efficiency. The stress-strain curve, characterizing global mechanical properties of a porous structure, could be well predicted (R(2)=0.92). Furthermore, validation of the method was achieved by comparing local displacements of element nodes with the displacements directly measured by time-lapsed imaging methods of failure, and these measures were in good agreement. The presented model is a first step in modeling specific samples for efficient strength analysis by FE modeling. We believe that with upcoming high-resolution in-vivo imaging methods, this approach could lead to a novel and accurate tool in the risk assessment for osteoporotic fractures.

Background: The impact of various medical and demographic factors on the quality of life (QoL) of breast cancer patients has been discussed controversially. We investigated the influence of six different factors on long-term QoL and body image of women with primary breast cancer., Patients and Methods: Two-hundred and seventy-four breast cancer patients were administered the QoL questionnaire following a mean interval of 4.2 years after primary diagnosis. All women had been primarily treated for stage I to III breast cancer without evidence of distant metastases. QoL was evaluated by using the QLQ-C30 questionnaire Version 2.0. Supplementary scales included body image, satisfaction with surgical treatment, cosmetic result and fear of recurrence. We analyzed the impact of tumor stage, surgical treatment, adjuvant radiotherapy, adjuvant cytotoxic therapy, age and length of follow-up period on the examined outcome parameters., Results: At the time of the follow-up examination, patients showed minor impairment of QoL (mean 67.8) and body image (mean 24.8), but more fear of recurrence (mean 60.7). None of the studied factors had a significant impact on overall QoL (P >0.05) according to the QLQ-C30 questionnaire. In contrast, with the exception of the factors 'cytotoxic therapy' and 'radiotherapy' all investigated variables influenced at least one of the additional psychological scales (P <0.05). The primary surgical treatment modality had the strongest impact and affected all four scales. Patients treated with breast conservation reported a more favorable body image, compared to those treated with mastectomy (17.2 versus 37.5, P <0.01), more satisfaction with surgical treatment (4.0 versus 10.7, P = 0.01), rated a better cosmetic result (75.5 versus 57.1, P <0.01), but presented more fear of recurrence (63.9 versus 55.3, P = 0.04)., Conclusion: Current QoL questionnaires do not sufficiently cover all relevant aspects of QoL, but might be complemented by breast cancer specific aspects such as body image and fear.

Clinical observations indicate that human immunodeficiency virus (HIV)-positive women experience more postoperative problems than do HIV-negative women. To obtain a better estimate of the individual risk of postoperative morbidity among HIV-infected women, and to determine which procedures pose the greatest risk, we performed a retrospective case-control study in which we assessed the outcomes after 235 obstetric and gynecologic surgical procedures. For purposes of comparison, an HIV-negative control patient was matched for each of the 235 surgical procedures performed, on the basis of the type of procedure and patient age. We found a significantly greater number of postoperative complications among the HIV-positive women. Higher complication rates occurred after abdominal surgery (odds ratio [OR], 3.6; P=.001) and curettage (OR, 7.7; P=.06). Among HIV-infected women, the risk of complications was associated with immune status. Antiretroviral therapy and standard perioperative antibiotic prophylaxis did not decrease the risk of complications. Indications for performing abdominal surgery and curettage on HIV-infected women should be carefully weighed against the potential risk of postoperative complications.

The members of the evolutionarily conserved Hox-gene complex, termed Hox genes, are required for specifying segmental identity during embryogenesis in various animal phyla. The Hox3 genes of winged insects have lost this ancestral function and are required for the development of extraembryonic epithelia, which do not contribute to any larval structure. Higher flies (Cyclorrhapha) such as Drosophila melanogaster contain Hox3 genes of two types, the zerknüllt type and the bicoid type. The zerknüllt gene is expressed zygotically on the dorsal side of the embryo and is required for establishing extraembryonic tissue. Its sister gene bicoid is expressed maternally and the transcripts are localized at the anterior pole of the mature egg. BICOID protein, which emerges from this localized source during early development, is required for embryonic patterning. All known direct bicoid homologues are confined to Cyclorrhaphan flies. Here, we describe Hox3 genes of the non-Cyclorrhaphan flies Empis livida (Empididae), Haematopota pluvialis (Tabanidae), and Clogmia albipunctata (Psychodidae). The gene sequences are more similar to zerknüllt homologues than to bicoid homologues, but they share expression characteristics of both genes. We propose that an ancestral Hox3 gene had been duplicated in the stem lineage of Cyclorrhaphan flies. During evolution, one of the gene copies lost maternal expression and evolved as zerknüllt, whereas the second copy lost zygotic expression and evolved as bicoid. Our finding correlates well with a partial reduction of zerknüllt-dependent extraembryonic tissue during Dipteran evolution.

In the last 10 years about 130 women with a drug addiction and more than 100 HIV-positive pregnant women were treated at the 1. University Hospital of Obstetrics and Gynaecology in Munich. Besides a specialized medical treatment both groups required intensive psychosocial care. HIV-infected people are still isolated and suffer from the social stigmata. Their essential needs for sexuality and children of their own are often ignored or even condemned because of irrational fears about HIV, which continue despite rapid medical improvements. The life-expectancy for example has increased since the inauguration of protease inhibitors. Vertical transmission of HIV is below 2% through medical treatment in pregnancy, elective cesarean section and renunciation of breastfeeding. Drug addicted pregnant women are given the opportunity to change their life in order to care for their children appropriately. The basis for this is a substitution with levomethadone and elimination of the use of other drugs. The addicted women often can reduce the dosage of levomethadone during the course of their pregnancy and sometimes can cease totally. Normally they are highly motivated and thus can ease the withdrawal symptoms of their newborns following delivery. By establishing a reliable social net during pregnancy mothers learn to recognize the demands of their children after birth and thus emotional and cognitive deficits can be prevented.

The Drosophila gene bicoid functions at the beginning of a gene cascade that specifies anterior structures in the embryo. Its transcripts are localized at the anterior pole of the oocyte, giving rise to a Bicoid protein gradient, which regulates the spatially restricted expression of target genes along the anterior-posterior axis of the embryo in a concentration-dependent manner. The morphogen function of Bicoid requires the coactivity of the zinc finger transcription factor Hunchback, which is expressed in a Bicoid-dependent fashion in the anterior half of the embryo. Whereas hunchback is conserved throughout insects, bicoid homologs are known only from cyclorrhaphan flies. Thus far, identification of hunchback and bicoid homologs rests only on sequence comparison. In this study, we used double-stranded RNA interference (RNAi) to address the function of bicoid and hunchback homologs in embryos of the lower cyclorrhaphan fly Megaselia abdita (Phoridae). Megaselia-hunchback RNAi causes hunchback-like phenotypes as observed in Drosophila, but Megaselia-bicoid RNAi causes phenotypes different from corresponding RNAi experiments in Drosophila and bicoid mutant embryos. Megaselia-bicoid is required not only for the head and thorax but also for the development of four abdominal segments. This difference between Megaselia and Drosophila suggests that the range of functional bicoid activity has been reduced in higher flies.

The Drosophila gene bicoid functions as the anterior body pattern organizer of Drosophila. Embryos lacking maternally expressed bicoid fail to develop anterior segments including head and thorax. In wild-type eggs, bicoid mRNA is localized in the anterior pole region and the bicoid protein forms an anterior-to-posterior concentration gradient. bicoid activity is required for transcriptional activation of zygotic segmentation genes and the translational suppression of uniformly distributed maternal caudal mRNA in the anterior region of the embryo. caudal genes as well as other homeobox genes or members of the Drosophila segmentation gene cascade have been found to be conserved in animal evolution. In contrast, bicoid homologs have been identified only in close relatives of the schizophoran fly Drosophila. This poses the question of how the bicoid gene evolved and adopted its unique function in organizing anterior-posterior polarity. We have cloned bicoid from a basal cyclorrhaphan fly, Megaselia abdita (Phoridae, Aschiza), and show that the gene originated from a recent duplication of the direct homolog of the vertebrate gene Hox3, termed zerknüllt, which specifies extraembryonic tissues in insects.

Course of Grief Response, influencing Factors, Satisfaction with treatment, Need for Psychotherapeutic Support: The emotional response to a late miscarriage or a stillbirth was examined retrospectively in a systematic study involving 51 patients. The women had lost a child either by stillbirth (beyond the 20th week of gestation) or shortly after delivery. We assessed the long-term course of possible determinants of grief, a complicated grief response, as well as the patients' satisfaction with the treatment and their need for further psychotherapeutic support. The mourning response was complicated by a depressive reaction in 22%. Major risk factors were individual disposition, lack of partner and professional support. The findings reveal that the course of the grief response is positively influenced by: 1. open discussion (thematisation) with the skilled personnel and the patients' families, 2. direct or indirect contact with the deceased child (concretisation) and 3. the patients concerned being extensively informed as to place and circumstances of the funeral (whereabouts of the child).

Only recently a stillbirth has been recognized as a major loss for the majority of women. Based on psychotherapeutic experience and clinical investigations, symptoms and risk factors of normal and pathological courses of grief are delineated and illustrated by a case vignette. Risk factors for complicated grief refer to quality of inpatient care, partner support, previous depression, anxiety or unresolved strain. Recommendations for inpatient care include validating the loss, making it real, enabling the bereaved parents to make informed decisions. Ambulatory after care should provide for monitoring and supporting the grief process.

In our opinion German gynaecology has failed to adequately face what came to pass during the Nazi period. This can be proved objectively, for there is no evidence that, after 1945, gynaecology had in any way cared to take notice--either thermatically or medically--of the thousands of victims of inhuman practices such as forced termination of pregnancy, compulsory sterilisation and the like. During the past 50 years recollections of enforced sterilisations, compulsory abortions, deliberate and hence criminal negligence and problematic approaches in research and teaching were almost completely banished from the area of conscious awareness and largely suppressed or silently ignored. Most of the medical directors of Departments of Gynaecology of German universities shared this view whenever they were questioned on the connections between gynaecology and Nazism. Now that two generations have passed it seems possible to examine and explore with less guilt feelings and shame the immensely fateful rôle of gynaecology in that context. Accent should be on the fate of the victims of that period. To bring back these events to memory, however, does not permit to conceal the part played by the physicians committing of these inhuman Nazi crimes. Data collected from a psychosomatically oriented examination of victims exemplify that to concretely recall gynaecology during Nazism a1-so offers a chance in several respects. One of the possibilities in this context is to signal "late apology" and regret to patients who had been victims, in one's own area of work, after one has psychically worked over their fate. Besides, a gynaecological-psychosomatic expertise will help e.g. that compulsorily sterilised women are granted financial aid that has at long last become a legal possibility and can be applied for since 1980. However, the relevant patient records do show very clearly that the inhuman practice of gynaecology during the so-called "Third Reich" was not only a collective problem but equally due to a failure of the individual conscience of numerous gynaecologists. Working over this complex may enhance our own sensitivity for psychosomatic and ethic problems and counteract any likelihood of a recurrence of an inhuman gynaecology.

Several more recent research results in psychosomatic obstetrics and gynecology will be presented, and directions for the practice will be given. In conclusion, it will be discussed how, for the realization of psychosomatic concepts, 'old-fashioned thinking' with biased, monocausal, scientific approaches is unproductive. This concerns, in particular, university gynecological hospitals where psychosomatic thinking is only very hesitantly accepted. The goal is, therefore, to promote a new thinking process, which incorporates an increased consideration of reference points like sensibility, flexibility and creativity.

Nowadays, in vitro fertilisation can be judged as a relatively successful treatment in special cases of sterility. Success cannot be defined only by induction of pregnancy, but must also be taken into consideration the couple's and the "IVF-children's" personal situation. The main problem of IVF is the high multiple rate of nearly 20%, resulting in a higher frequency of preterm deliveries. This leads to higher perinatal mortality and more longterm deficiencies in these children. The women's attitude during pregnancy can be considered to be responsible. From a psychosomatic point of view (the husband's presence during delivery, breast feeding etc.) there are favourable preconditions for the "birth of the family." In most cases, the couples give a contented or good view of their partnership (self-assessment). Nearly 5% of them seem to be "disappointed" after the birth of the baby. The method of IVF is positively assessed from an overall point of view. But only half of the women do not keep this special method of procreation secret.

It is well known that tendon or muscle interposition can cause fractures to be irreducible by closed means. A rare tendon interposition of extensor carpi ulnaris occurred in an 11-year-old boy. The isolated displaced distal ulnar physeal injury was the result of a motor vehicle accident. Attempts to perform a closed reduction failed. At operation, an interposed extensor carpi ulnaris tendon was found. After repositioning the tendon, a near anatomic reduction was easily accomplished. Only two cases seem to have been reported in the literature on the need for open reduction of distal ulnar physeal injury, none with an interposed tendon. Interposed extensor carpi ulnaris tendon should be considered as a cause of irreducible displaced distal ulna epiphyseal fracture.

Nine pregnant women with the AIDS problem were under treatment and care at the Department of Gynaecology of the University of Berlin at Charlottenburg during 1985. Of these, seven had HTLV-III antibodies only, whereas one woman showed additional clinical signs such as lymphoadenopathy and opportunistic infections; another woman developed exanthemas during pregnancy that pointed to a possible manifestation of AIDS. The increasing incidence in our clinic of pregnant women infected with or actually suffering from AIDS is due to the fact that for several years we have been conducting heroin withdrawal programmes in pregnant women. It must be born in mind, however, that generally more and more women outside the high-risk groups covered by anamnesis are being infected with HTLV-III. Screening tests for HTLV-III can help to clarify matters and make it possible to care adequately for the patient and to guarantee the obstetric team an appropriate management of the risk of infection. However, it must be considered as a minimum demand to be made on the obstetrician to conduct tests for HTLV-III antibodies in women of the high-risk groups (heroin addicts, prostitutes, blood transfusion recipients, fathers at risk). Based on update knowledge, the authors supply pointers to a more realistic information of pregnant women. Suggestions are advanced in respect of care during pregnancy, childbirth and puerperium.

The mounting problems of drug addiction now also confront the obstetrician and pediatrician. 1/3 of the 60,000 addicts to opiates in the Federal Republic of Germany are women. Of these 80% are in the reproductive age group. Despite endocrine problems pregnancies do occur and are high risk pregnancies because of hepatitis, venereal disease, malnutrition, phlebitis, abscesses, premature deliveries, premature rupture of the membranes, malpresentations, dysmaturity, pre-eclampsia, and numerous other social and psychological problems. Because of the varying content of heroin on the black market the fetus runs the risk of acute overdose or withdrawal. Withdrawal results in extremely marked fetal movements, with increased oxygen consumption and a danger of intra-uterine asphyxia sometimes resulting in intra-uterine fetal death. Most newborns develop a withdrawal syndrome with irritability, high pitched cries, shivering, tachycardia, perspirations, fever and generalized seizures. The experience of the Department for Women for the Free University in Berlin-Charlottenburg are presented and compared to a review of the literature. The medical, social, and psychological aspects of the problem are discussed.

Chromosomal abnormalities and abnormal embryonic development have been observed after human in vitro fertilisation (IVF). Such investigations are not permitted according to the ethical guidelines approved by the IVF team in Berlin ("Berliner Modell"). However, the chromosomal status was studied in oocytes which remained unfertilised in a human IVF programme. In 50 fertilisation attempts the chromosomes of 62 oocytes could be analysed. 45 of the oocytes were in the process of meiotic maturation. One of these showed abnormalities in the first meiotic division and seven during the second meiotic division. After fertilisation in addition to normal embryos, all of which were transferred to the mother, three embryos were triploid and six oocytes were fragmented. The results prove that in human IVF chromosome abnormalities can occur in oocytes already before fertilisation. The importance of the results for later stages of embryonic development is discussed.