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Immunology of pregnancy. Implications for the mother.

Pregnancy is a unique event in which a genetically and immunologically foreign fetus usually survives to full term without apparent rejection by the mother's immune system. Over the past decade, more information has been gathered to provide insight into the complex immunological mechanisms that allow the fetus to grow and survive in most cases. Whereas the maternal-fetal interface was once felt to be an immunologically privileged site with complete separation between the fetus and the mother, it is now known that recognition of the foreign fetus does occur. However, despite this immunological recognition, several mechanisms have been discovered which may explain why the mother does not reject the foreign fetus. These mechanisms include fetal factors such as trophoblast cell properties and altered MHC Class I expression as well as local maternal factors such as specialized uterine natural killer cells and a shifting of the T-helper cell cytokine profile from a type 1 to a type II array. Other novel immunomodulators are found to be expressed in the local uterine environment to aid in fetal survival. Furthermore, the persistence of fetal cells in the maternal circulation long after pregnancy is over (termed chronic microchimerism) and may have implications for autoimmune diseases. This review presents investigations and developments relevant to an understanding as to why the fetus is not rejected by the maternal immune system.

Cytokines↗

Multilingual biomedical dictionary.

We present a unique technique to create a multilingual biomedical dictionary, based on a methodology called Morpho-Semantic indexing. Our approach closes a gap caused by the absence of free available multilingual medical dictionaries and the lack of accuracy of non-medical electronic translation tools. We first explain the underlying technology followed by a description of the dictionary interface, which makes use of a multilingual subword thesaurus and of statistical information from a domain-specific, multilingual corpus.

Abstracting and Indexing↗

Infiltration of hypertrophic esophageal smooth muscle by mast cells and basophils.

Partial obstruction leads to chronic distension and muscular hypertrophy of the opossum esophagus. The smooth muscle cells of the circular muscle layer enlarge, become pleomorphic and are surrounded by an amorphous ground substance in the extracellular space. Here we describe the histological and ultrastructural features of a peculiar cellular infiltrate in the hypertrophic smooth muscle. The infiltrate consisted uniquely of mast cells and basophils. In per unit area, the number of mast cells increased from 0.9 +/- 0.1 cells in controls to 3.7 +/- 0.2 in hypertrophic smooth muscle; the corresponding numbers for basophils were 2.5 +/- 0.2 and 7.2 +/- 0.3 cells. Cells were seen primarily in the septal spaces of the circular muscle layer and at the interface of the circular and longitudinal muscle layer. The cytoplasm of basophils is normally packed with round and oval granules. The granules stain metachromatically and with varying intensity on Wright-Giemsa stains. On transmission electronmicroscopy, granules display a membrane and a great diversity in the structure of their luminal contents. In hypertrophic muscle, most granules were discharging their contents into the cytoplasm or extracellular space. The membranes of adjacent empty granules then fused to form a chain of vacuoles. Similar changes occurred also in the mast cells which differed from the basophil by their lack of nuclear lobulation and by the greater homogeneity of their cytoplasmic granules. It is possible that these inflammatory cells are involved in the reconstruction of the smooth muscle and its connective tissue which occur during esophageal distension and hypertrophy.

Animals↗

Distribution of CD14+ and CD68+ macrophages in the placental bed and basal plate of women with preeclampsia and preterm labor.

OBJECTIVE: Macrophages play a key role in implantation, placentation and parturition. Yet, whether or not the number of macrophages at the fetomaternal interface (basal plate of the placenta and placental bed) is altered in women with preeclampsia is the subject of controversy. The purpose of this study was to compare the immunoreactivity and distribution patterns of CD14 and CD68 positive macrophages in both the basal plate and placental bed from preeclamptic and non-preeclamptic pregnancies. METHODS: A cross-sectional study was conducted. Paraffin embedded sections of placental tissues and placental bed biopsies were obtained from patients with early onset preeclampsia (n=10) and from those with preterm labor/delivery (n=10) without preeclampsia matched for gestational age. Double immunohistochemistry using antibodies to CD14 and CD68 was performed, and the density of double or single positive cells in the basal plate and placental bed was evaluated. Non-parametric statistics were used for analysis. RESULTS: 1) A unique subset of CD14-/CD68+ cells was identified. The cells in question were present at a higher level in the decidua than in the myometrial segment of the placental bed (p<0.01); 2) The density and proportion of CD14+/CD68+ cells (double positive cells) were significantly higher in the myometrial segment than in the basal plate (p=0.0003); and 3) There were no significant differences in the density and patterns of immunopositive macrophages in the basal plate, the decidua, and the myometrium between women with preeclampsia and those with preterm labor/delivery (p>0.05). CONCLUSION: The macrophages at the fetomaternal interface can be dichotomized by CD14 and CD68 immunoreactivity. A gradient of CD14+/CD68+ macrophages was demonstrated between the superficial myometrium and the basal plate regardless of the etiology of preterm birth (preeclampsia or spontaneous preterm labor). The biological function of single positive (CD14-/CD68+) and double positive (CD14+/CD68+) macrophages at the fetomaternal interface remains to be established. The overall findings also suggest that the discrepancies in the literature are due to the varying markers used to detect macrophages and in the anatomical plane of the fetomaternal junction analyzed.

Antigens, CD↗

Melanie II--a third-generation software package for analysis of two-dimensional electrophoresis images: I. Features and user interface.

Although two-dimensional electrophoresis (2-DE) computer analysis software packages have existed ever since 2-DE technology was developed, it is only now that the hardware and software technology allows large-scale studies to be performed on low-cost personal computers or workstations, and that setting up a 2-DE computer analysis system in a small laboratory is no longer considered a luxury. After a first attempt in the seventies and early eighties to develop 2-DE analysis software systems on hardware that had poor or even no graphical capabilities, followed in the late eighties by a wave of innovative software developments that were possible thanks to new graphical interface standards such as XWindows, a third generation of 2-DE analysis software packages has now come to maturity. It can be run on a variety of low-cost, general-purpose personal computers, thus making the purchase of a 2-DE analysis system easily attainable for even the smallest laboratory that is involved in proteome research. Melanie II 2-D PAGE, developed at the University Hospital of Geneva, is such a third-generation software system for 2-DE analysis. Based on unique image processing algorithms, this user-friendly object-oriented software package runs on multiple platforms, including Unix, MS-Windows 95 and NT, and Power Macintosh. It provides efficient spot detection and quantitation, state-of-the-art image comparison, statistical data analysis facilities, and is Internet-ready. Linked to proteome databases such as those available on the World Wide Web, it represents a valuable tool for the "Virtual Lab" of the post-genome area.

Algorithms↗

Modeling the nuclear magnetic resonance behavior of lung: from electrical engineering to critical care medicine.

The present article reviews the basic principles of a new approach to the characterization of pulmonary disease. This approach is based on the unique nuclear magnetic resonance (NMR) properties of the lung and combines experimental measurements (using specially developed NMR techniques) with theoretical simulations. The NMR signal from inflated lungs decays very rapidly compared with the signal from completely collapsed (airless) lungs. This phenomenon is due to the presence of internal magnetic field inhomogeneity produced by the alveolar air-tissue interface (because air and water have different magnetic susceptibilities). The air-tissue interface effects can be detected and quantified by magnetic resonance imaging (MRI) techniques using temporally symmetric and asymmetric spin-echo sequences. Theoretical models developed to explain the internal (tissue-induced) magnetic field inhomogeneity in aerated lungs predict the NMR lung behavior as a function of various technical and physiological factors (e.g., the level of lung inflation) and simulate the effects of various lung disorders (in particular, pulmonary edema) on this behavior. Good agreement has been observed between the predictions obtained from the mathematical models and the results of experimental NMR measurements in normal and diseased lungs. Our theoretical and experimental data have important pathophysiological and clinical implications, especially with respect to the characterization of acute lung disease (e.g., pulmonary edema) and the management of critically ill patients.

Blood-Air Barrier↗

Recent advances in mycotoxin determination in food and feed by hyphenated chromatographic techniques/mass spectrometry.

Mycotoxins are fungal toxins produced by molds, which occur universally in food and feed derivatives, and are produced under certain environmental conditions in the field before harvest, post-harvest, during storage, processing, and feeding. Mycotoxin contamination is one of the most relevant and worrisome problem concerning food and feed safety because it can cause a variety of toxic acute and chronic effects in human and animals. In this review we report the use of mass spectrometry in connection with chromatographic techniques for mycotoxin determination by considering separately the most diffuse class of mycotoxins: patulin, aflatoxins, ochratoxin A, zearalenone, trichothecenes, and fumonisins. Although the selectivity of mass spectrometry is unchallenged if compared to common GC and LC detection methods, accuracy, precision, and sensitivity may be extremely variable concerning the different mycotoxins, matrices, and instruments. The sensitivity issue may be a real problem in the case of LC/MS, where the response can be very different for the different ionization techniques (ESI, APCI, APPI). Therefore, when other detection methods (such as fluorescence or UV absorbance) can be used for the quantitative determination, LC/MS appears to be only an outstanding confirmatory technique. In contrast, when the toxins are not volatile and do not bear suitable chromophores or fluorophores, LC/MS appears to be the unique method to perform quantitative and qualitative analyses without requiring any derivatization procedure. The problem of exact quantitative determination in GC/MS and LC/MS methods is particularly important for mycotoxin determination in food, given the high variability of the matrices, and can be solved only by the use of isotopically labeled internal standards or by the use of ionization interfaces able to lower matrix effects and ion suppressions. When the problems linked to inconstant ionization and matrix effects will be solved, only MS detectors will allow to simplify more and more the sample preparation procedures and to avoid clean-up procedures, making feasible low-cost, high-throughput determination of mycotoxins in many different food matrices.

Animal Feed↗

Crystal structure of putidaredoxin, the [2Fe-2S] component of the P450cam monooxygenase system from Pseudomonas putida.

Stability of the [2Fe-2S]-containing putidaredoxin (Pdx), the electron donor to cytochrome P450cam in Pseudomonas putida, was improved by mutating non-ligating cysteine residues, Cys73 and Cys85, to serine singly and in combination. The increasing order of stability is Cys73Ser/Cys85Ser>Cys73Ser>Cys85Ser>WT Pdx. Crystal structures of Cys73Ser/Cys85Ser and Cys73Ser mutants of Pdx, solved by single-wavelength anomalous dispersion phasing using the [2Fe-2S] iron atoms to 1.47 A and 1.65 A resolution, respectively, are nearly identical and very similar to those of bovine adrenodoxin (Adx) and Escherichia coli ferredoxin. However, unlike the Adx structure, no motion between the core and interaction domains of Pdx is observed. This higher conformational stability of Pdx might be due to the presence of a more extensive hydrogen bonding network at the interface between the two structural domains around the conserved His49. In particular, formation of a hydrogen bond between the side-chain of Tyr51 and the carbonyl oxygen atom of Glu77 and the presence of two well-ordered water molecules linking the interaction domain and the C-terminal peptide to the core of the molecule are unique to Pdx. The folding topology of the NMR model is similar to that of the X-ray structure of Pdx. The overall rmsd of Calpha positions between the two models is 1.59 A. The largest positional differences are observed for residues 18-21 and 33-37 in the loop regions and the C terminus. The latter two peptides display conformational heterogeneity in the crystal structures. Owing to flexibility, the aromatic ring of the C-terminal Trp106 can closely approach the side-chains of Asp38 and Thr47 (3.2-3.9 A) or move away and leave the active site solvent-exposed. Therefore, Trp106, previously shown to be important in the Pdr-to-Pdx and Pdx-to-P450cam electron transfer reactions is in a position to regulate and/or mediate electron transfer to or from the [2Fe-2S] center of Pdx.

Adrenodoxin↗

Neuropeptide Y-containing neurons are situated predominantly outside cytochrome oxidase puffs in macaque visual cortex.

Layers II/III of the primary visual cortex contain a regular pattern of histochemically detectable cytochrome oxidase (CO)-rich "puffs," which differ from the interpuff regions in their thalamo-cortical and cortico-cortical connectivity, receptive-field properties, and the density of inhibitory GABA-containing synaptic terminals. We used an immunocytochemical method, in combination with cytochrome oxidase histochemistry, to analyze the spatial relationship between neurons that contain neuropeptide Y (NPY) and the CO puffs. Of a total of 606 neurons, only 2.6% of the NPY-containing cells are located in the puffs, whereas the rest are situated in the interpuffs, or at the interface between puffs and interpuffs. The number of NPY-containing neurons in the puffs is substantially less than that expected in an equal volume of the interpuffs (X2 = 13.86; df = 1; P less than 0.001). These observations indicate that columns containing the puffs may differ also from those in the interpuff regions in that they contain a unique array of chemically and morphologically distinct local circuit neurons.

Animals↗

Strategies for chemical reaction searching in SciFinder

The bibliographic, chemical structure, and chemical reaction databases produced by Chemical Abstracts Service allow a number of possibilities for chemical reaction searching. While these same databases may be searched through the STN network, many end-users find the intuitive software interface SciFinder simpler, but there still are issues to address. Searching may be performed through keywords, chemical structures, or chemical reactions, and the answers may vary with respect to precision and comprehension. Often combinations of search options may be needed to best solve the problem. Retrosynthetic analyses are easily performed in the chemical reaction database and can give unique insights into synthetic alternatives.

Journal Article↗

Phage P4 origin-binding domain structure reveals a mechanism for regulation of DNA-binding activity by homo- and heterodimerization of winged helix proteins.

The origin-binding domain of the gpalpha protein of phage P4 (P4-OBD) mediates origin recognition and regulation of gpalpha activity by the protein Cnr. We have determined the crystal structure of P4-OBD at 2.95 A resolution. The structure of P4-OBD is that of a dimer with pseudo twofold symmetry. Each subunit has a winged helix topology with a unique structure among initiator proteins. The only structural homologue of the P4-OBD subunit is the DNA-binding domain of the eukaryotic transcriptional activator Rfx1. Based on this structural alignment, a model for origin recognition by the P4-OBD dimer is suggested. P4-OBD mutations that interfere with Cnr binding locate to the dimer interface, indicating that Cnr acts by disrupting the gpalpha dimer. P4-OBD dimerization is mediated by helices alpha1 and alpha3 in both subunits, a mode of winged helix protein dimerization that is reminiscent of that of the eukaryotic transcription factors E2F and DP. This, in turn, suggests that Cnr is also a winged helix protein, a possibility that is supported by previously unreported sequence homologies between Cnr and Rfx1 and homology modelling. Hence, in a mechanism that appears to be conserved from phage to man, the DNA-binding activity of winged helix proteins can be regulated by other winged helix proteins via the versatile use of the winged helix motif as a homo- or heterodimerization scaffold.

Amino Acid Sequence↗

Crystal structure at 2.6-A resolution of human macrophage migration inhibitory factor.

Macrophage migration inhibitory factor (MIF) was the first cytokine to be described, but for 30 years its role in the immune response remained enigmatic. In recent studies, MIF has been found to be a novel pituitary hormone and the first protein identified to be released from immune cells on glucocorticoid stimulation. Once secreted, MIF counterregulates the immunosuppressive effects of steroids and thus acts as a critical component of the immune system to control both local and systemic immune responses. We report herein the x-ray crystal structure of human MIF to 2.6 angstrom resolution. The protein is a trimer of identical subunits. Each monomer contains two antiparallel alpha-helices that pack against a four-stranded beta-sheet. The monomer has an additional two beta-strands that interact with the beta-sheets of adjacent subunits to form the interface between monomers. The three beta-sheets are arranged to form a barrel containing a solvent-accessible channel that runs through the center of the protein along a molecular 3-fold axis. Electrostatic potential maps reveal that the channel has a positive potential, suggesting that it binds negatively charged molecules. The elucidated structure for MIF is unique among cytokines or hormonal mediators, and suggests that this counterregulator of glucocorticoid action participates in novel ligand-receptor interactions.

Amino Acid Sequence↗

An industry, clinical, and academic telehealth partnership venture: progress, goals achieved, and lessons learned.

The goal of this project was to develop and test a multi-application telehealth workstation with an interface seamless to the end-users. This project required collaboration among the private sector, a clinical regional referral setting, a clinical tertiary receiving center, and a university academic unit. The project applied the usability testing methodology to design and test the workstation: (1) the developmental phase focused on planning, prototype workstation development, and end-user trials in the private sector research and development environment, and (2) the operational testing phase moved the workstation into the clinical setting. The latter included training of end-users and addressing policy and ethical issues. In addition, the partners documented the goals achieved and lessons learned. The project resulted in the refinement of the workstation for clinical applications. The unique, diverse, and at times complementary goals and lessons learned by each partner are noted. Collaboration around a shared goal was a key element in achieving the refined workstation. Collaborators reported that the lessons learned will aid them as they pursue telehealth activities.

Alberta↗

Developing a coding scheme for detecting usability and fun problems in computer games for young children.

This article describes the development and assessment of a coding scheme for finding both usability and fun problems through observations of young children playing computer games during user tests. The proposed coding scheme is based on an existing list of breakdown indication types of the detailed video analysis method (DEVAN). This method was developed to detect usability problems in task-based products for adults. However, the new coding scheme for children's computer games takes into account that in games, fun, in addition to usability, is an important factor and that children behave differently from adults. Therefore, the proposed coding scheme uses 8 of the 14 original breakdown indications and has 7 new indications. The article first discusses the development of the new coding scheme. Subsequently, the article describes the reliability assessment of the coding scheme. The any-two agreement measure of 38.5% shows that thresholds for when certain user behavior is worth coding will be different for different evaluators. However, the any-two agreement of .92 for a fixed list of observation points shows that the distinction between the available codes is clear to most evaluators. Finally, a pilot study shows that training can increase any-two agreement considerably by decreasing the number of unique observations, in comparison with the number of agreed upon observations.

Child↗

The hospital library online--a point of service for consumers and hospital staff: a case study.

The Health Library at Stanford University is described in the context of electronic information services provided to Stanford University Medical Center, the local community, and Internet users in general. The evolution from CD-ROM-based services to Web-based services and in-library services to networked resources are described. Electronic services have expanded the mission of The Health Library to include national and international users and the provision of unique services and collections.

CD-ROM↗

Structural and functional organization of a diencephalic sensory-motor interface in the gymnotiform fish, Eigenmannia.

The diencephalic nucleus electrosensorius (nE) of gymnotiform fish comprises a series of finely tuned neuronal filters for control of the jamming avoidance response (JAR) and probably other electromotor tasks as well. The nE receives electrosensory input from the dorsal torus semicircularis (TSd) and octavolateral input from the ventral torus (TSv). The nE, in turn, projects to various hypothalamic and thalamic nuclei, including the prepacemaker nucleus (PPn), which can modulate the frequency of electric organ discharges (EODs) via its unique input to the medullary pacemaker nucleus. Four subdivisions of the nE can now be recognized: 1) The beat-related area (nEb)--a rostral cluster of tightly packed cells which receives TSd input and projects to the inferior lobe, anterior tuberal nucleus, anterior thalamic nucleus, central posterior thalamic nucleus, and PPn. The nEb contains neurons responsive to beat patterns caused by jamming stimuli. Stimulation of the nEb with L-glutamate, however, fails to induce any EOD-frequency shift. 2) The area causing EOD-frequency rises (nE increases)--a horizontal band of cells at the dorsal aspect of the caudal nE which receives TSd input and projects to the PPn and vicinity and to the cerebellum; nE increases stimulation induces slow EOD-frequency rises characteristic of the JAR. Responses of these cells to jamming stimuli are not yet known. 3) The area causing EOD-frequency falls (nE decreases)--a horizontal band of cells at the ventral aspect of the caudal nE which receives TSd input and projects only to the PPn and vicinity; nE decreases stimulation induces slow EOD-frequency falls characteristic of the JAR. The responses of these cells to jamming stimuli are not yet known. 4) The acousticolateral region (nEar)--a complex medial region of the nE which receives input predominantly from the ventral torus and projects to the inferior lobe, anterior tuberal nucleus, central posterior thalamic nucleus, PPn, and cerebellum; the sensory and motor properties of this region are not known in detail, although auditory and mechanosensory responses have been recorded here. Projections to the PPn and its vicinity suggest direct control of electromotor behaviors by the nE, whereas thalamic and hypothalamic projections may provide a substrate for electrosensory influences on neuroendocrine and motivational control centers. The optic tectum projects strongly to the pretectum and various other diencephalic nuclei in the vicinity of the nE, but it does not innervate the nE itself. Accordingly, ablation of the tectum does not affect the performance of the JAR.

Animals↗

Collaborative learning using Internet2 and remote collections of stereo dissection images.

We have investigated collaborative learning of anatomy over Internet2, using an application called remote stereo viewer (RSV). This application offers a unique method of teaching anatomy, using high-resolution stereoscopic images, in a client-server architecture. Rotated sequences of stereo image pairs were produced by volumetric rendering of the Visible female and by dissecting and photographing a cadaveric hand. A client-server application (RSV) was created to provide access to these image sets, using a highly interactive interface. The RSV system was used to provide a "virtual anatomy" session for students in the Stanford Medical School Gross Anatomy course. The RSV application allows both independent and collaborative modes of viewing. The most appealing aspects of the RSV application were the capacity for stereoscopic viewing and the potential to access the content remotely within a flexible temporal framework. The RSV technology, used over Internet2, thus serves as an effective complement to traditional methods of teaching gross anatomy.

Anatomy↗

A signaling role for the uncleaved form of alpha 6 integrin in differentiating lens fiber cells.

Many alpha integrin subunits are cleaved during their processing to yield heavy and light chains, which remain associated by disulfide bonds. While uncleaved alpha integrin subunits can form functional receptors that sometimes have distinct signaling roles from their better-characterized endoproteolytically cleaved counterparts, their expression at the cell surface and their association with signaling complexes have yet to be determined in vivo. In this study, we demonstrate that, in differentiating lens fiber cells, the uncleaved form of alpha 6 integrin was expressed at the cell surface. This form of alpha 6 integrin coimmunoprecipitated with both the signaling adaptor molecule Shc and its downstream effector Grb2, suggesting that, in lens fiber cells, uncleaved alpha 6 integrin was associated with a Shc-mediated signaling complex. We show that expression of the cleaved form of alpha 6 integrin progressively decreased relative to uncleaved alpha 6 integrin as the state of lens cell differentiation increased, resulting in the predominance of uncleaved alpha 6 integrin in the lens fiber cell zones. Interestingly, we previously have shown that alpha 6 integrin is localized principally along the extensive cell-cell interfaces of these lens fiber cells, in the absence of its extracellular matrix ligand laminin. While we found that the cleaved form of alpha 6 integrin contained both high mannose and complex sugars, the uncleaved form of alpha 6 integrin contained only high mannose sugars. These properties suggest that the uncleaved form of alpha 6 integrin may have a unique role in the embryonic lens. Its high association with Shc and Grb2 in the differentiating cortical fiber cell zone indicates that alpha 6 integrin may provide a cell survival signal in the presence of the apoptotic-like processes that are initiated in this region of the embryonic lens to clear the lens cells of their organelles.

Adaptor Proteins, Signal Transducing↗