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An economised craniofacial identification system.

It has been attempted to develop an economised craniofacial identification system, as a special automated version of photo/video superimposition technique, that can deal with common cases of personal identification with the aid of a skull and a nearly front view face photograph of the suspected victim. The proposed method is economic in respect of (i) cost of hardware configuration, (ii) processing time as well as (iii) manual labour involved. Over and above, it has got a capability to take care of ambiguities due to soft tissue thickness during the selection of facial features, which is a part of the procedure. In order to reconstruct a 2-D cranial image, superimposable over the facial one, the new method does not need any reconstruction of a digitised 3-D cranial image. It works simply by a suitable segment-wise processing of a 2-D cranial image with the aid of the symmetry perceiving adaptive neuronet (SPAN), that has recently been introduced in connection with nearly front view facial image recognition. The final comparison of the facial and the superimposable cranial images is as versatile as the same for facial image recognition by SPAN.A practical application of this extended version of SPAN has been demonstrated in the present paper.

Child↗

Self-organizing continuous attractor networks and motor function.

Motor skill learning may involve training a neural system to automatically perform sequences of movements, with the training signals provided by a different system, used mainly during training to perform the movements, that operates under visual sensory guidance. We use a dynamical systems perspective to show how complex motor sequences could be learned by the automatic system. The network uses a continuous attractor network architecture to perform path integration on an efference copy of the motor signal to keep track of the current state, and selection of which motor cells to activate by a movement selector input where the selection depends on the current state being represented in the continuous attractor network. After training, the correct motor sequence may be selected automatically by a single movement selection signal. A feature of the model presented is the use of 'trace' learning rules which incorporate a form of temporal average of recent cell activity. This form of temporal learning underlies the ability of the networks to learn temporal sequences of behaviour. We show that the continuous attractor network models developed here are able to demonstrate the key features of motor function. That is, (i) the movement can occur at arbitrary speeds; (ii) the movement can occur with arbitrary force; (iii) the agent spends the same relative proportions of its time in each part of the motor sequence; (iv) the agent applies the same relative force in each part of the motor sequence; and (v) the actions always occur in the same sequence.

Excitatory Postsynaptic Potentials↗

Mouse visual cortex.

Neurons in mouse visual cortex have diverse receptive field properties and they respond selectively to specific features of visual stimuli. Owing to the lateral position of the eyes, only about a third of the visual cortex receives input from both eyes, but many cells in this region are binocular. Similar to higher mammals, closing one eye during a critical period shifts the responses of cells, such that they are better driven by the non-deprived eye. In this review I illustrate how the combination of transgenic mouse technology with single cell recording and modern imaging techniques might lead to a further understanding of the mechanisms that underlie the development, plasticity, and function of the mammalian visual cortex.

Animals↗

Causes of inconsistency in diagnosing and classifying intraductal proliferations of the breast. European Commission Working Group on Breast Screening Pathology.

It is now widely recognised that classifying ductal carcinoma in situ (DCIS) of the breast and diagnosing atypical ductal hyperplasia are associated with significant interobserver variation. Two possible reasons for this inconsistency are differences in the interpretation of specified histological features and field selection where morphology is heterogeneous. In order to investigate the relative contribution of these two factors to inconsistent interpretation of intraductal proliferations, histological sections of 32 lesions were sent to 23 European pathologists followed 3 years later by images of small parts of these sections. Kappa statistics for diagnosing hyperplasia of usual type, atypical ductal hyperplasia and ductal carcinoma in situ were 0.54, 0.35 and 0.78 for sections and 0.47, 0.29 and 0.78 for images, respectively, showing that most of the inconsistency is due to differences in morphological interpretation. Improvements can thus be expected only if diagnostic criteria or methodology are changed. In contrast, kappa for classifying DCIS by growth pattern was very low at 0.23 for sections and better at 0.47 for images, reflecting the widely recognised variation in the growth pattern of DCIS. Higher kappa statistics were obtained when any mention of an individual growth pattern was included in that category, thus allowing multiple categories per case; but kappa was still higher for images than sections. Classifying DCIS by nuclear grade gave kappa values of 0.36 for sections and 0.49 for images, indicating that intralesional heterogeneity has hitherto been underestimated as a cause of inconsistency in classifying DCIS by this method. More rigorous assessment of the proportions of the different nuclear grades present could lead to an improvement in consistency.

Breast↗

Fast-tracking revisited: routine cardiac surgical patients need minimal intensive care.

OBJECTIVE: Following cardiac surgery, patients are transferred from the operating theatre to intensive care. This clinical environment has one nurse per patient and facilities for mechanical ventilation. Patients are kept in this setting until the following day. This practice has been challenged with early extubation of patients. At our institution we have established a fast-track policy including the following features: (1) patient selection; (2) operating list scheduling with fast-track patients first; (3) anaesthetic tailored to early extubation; (4) methodical procedure with warm cardiopulmonary bypass; (5) removal of the arterial line; (6) transfer from intensive care to a separate high dependency unit ('step-down') on the day of operation, where the ratio of nurse to patient is one to three and there are no ventilatory facilities and no invasive monitoring; or (7) to keep these patients on ICU but decrease the nurse to patient ratio. METHOD: The case notes of 572 patients who predominantly had myocardial revascularisation, undergoing this process from July 1996 to July 2000 at our institution were reviewed. RESULTS: Mean EUROSCORE for the study group was 1.42. The 30-day mortality rate for the study group was 0.34%, mean intensive care time was 5 h 52 min, mean time to extubation was 3 h 10 min, mean readmission rate to intensive care was 0.34% and mean hospital stay from day of operation (inclusive) was 5.65 days. This process increased our throughput by 14.6% (compared to standard practices). COMMENT: This study demonstrates that transfer of appropriate patients to a high dependency area from intensive care following cardiac surgery is safe. It allows intensive care beds to be used by more than one patient each day and allows significant cost savings by reducing the nursing ratio per patient.

Adult↗

Cellular survival pathways and resistance to cancer therapy.

Chemotherapy and irradiation induce programmed cell death (apoptosis) in their target cells. Dysregulated apoptosis is a feature that is selected for during tumor formation and contributes to therapeutic resistance. Cell survival in the face of cytotoxic therapy is dictated by both internal properties of the cell, such as status of components of the apoptotic machinery, and its extracellular milieu, such as extracellular matrix (ECM) and growth factor receptor expression and signaling. The importance of extracellular survival signals as key regulators of apoptosis is now being recognized by the ability of growth factors (GFs), GF receptors (GFRs), and GFR signaling to promote cellular survival. GFs can mitigate or abrogate the effectiveness of cancer therapy and protect against other cellular insults. Because survival signals generated by different extracellular sources converge at key molecules, new molecular targets have been identified which could be exploited to maximize the effectiveness of cytotoxic cancer therapy.

Journal Article↗

Processing of local and global visual features in obsessive-compulsive disorder.

Research conducted with the Rey-figure task has suggested that patients with obsessive-compulsive disorder (OCD) process local aspects of a complex stimulus more efficiently than the overall gestalt. The aim of the present study was to investigate if this "local bias" is established already during early stimulus encoding or occurs only during later processing, once a percept has been formed (e.g., memory retrieval). To this end, responding to local and global targets of hierarchical letters (e.g., an "E" composed out of small "T"s) was assessed in 30 OCD patients and 28 healthy controls. OCD patients and controls performed comparably on all parameters. These results lend no support to the notion of an early perceptual bias towards local elements in OCD patients. It remains to be tested whether a bias towards local features is confined to situations where local and global features compete for selection.

Adult↗

Differential mass spectrometry: a label-free LC-MS method for finding significant differences in complex peptide and protein mixtures.

Efficiently identifying and quantifying disease- or treatment-related changes in the abundance of proteins is an important area of research for the pharmaceutical industry. Here we describe an automated, label-free method for finding differences in complex mixtures using complete LC-MS data sets, rather than subsets of extracted peaks or features. The method selectively finds statistically significant differences in the intensity of both high-abundance and low-abundance ions, accounting for the variability of measured intensities and the fact that true differences will persist in time. The method was used to compare two complex peptide mixtures with known peptide differences. This controlled experiment allowed us to assess the validity of each difference found and so to analyze the method's sensitivity and specificity. The method detects both presence versus absence and a 2-fold change in peptide concentration near the limit of detection of the instrument used, where chromatographic peaks may not be sufficiently well defined to be detected in individual samples. The method is more sensitive and gives fewer false positives than subtractive methods that ignore signal variability. Differential mass spectrometry combined with targeted MS/MS analysis of only identified differences may save both computation time and human effort compared to shotgun proteomics approaches.

Chromatography, Liquid↗

(1S)-1-(aminomethyl)-2-(arylacetyl)-1,2,3,4-tetrahydroisoquinoline and heterocycle-condensed tetrahydropyridine derivatives: members of a novel class of very potent kappa opioid analgesics.

The synthesis and structure-activity relationship (SAR) of a novel class of kappa opioid analgesics, 1-(aminomethyl)-2-(arylacetyl)-1,2,3,4- tetrahydroisoquinolines and (aminomethyl)-N-(arylacetyl)-4,5,6,7-tetrahydrothienopyridines+ ++, are described. These compounds, formally derived by the condensation of a benzene or thiophene ring on the piperidine nucleus of the recently described compounds 1, are from 3 to 7 times more potent as antinociceptive agents and with a longer duration of action than the original lead compounds. A similar N2-C1-C9-N10 pharmacophore torsional angle of approximately 60 degrees was also found for this class of compounds by using X-ray and 1H NMR analyses. The same absolute configuration (S) at the chiral center of the active (-) enantiomers was determined by X-ray crystallographic analysis. A varied degree of kappa receptor selectivity was a feature of this novel class of antinociceptive agents (mu/kappa ratio from 44 to 950 according to the nature of the basic moiety). SAR analysis indicated that the presence of electron-withdrawing and lipophilic substituents in para and/or meta positions in the arylacetic moiety and the pyrrolidino or dimethylamino basic groups are required to optimize biological activity. The lead compounds 28, 30, and 48 are among the most potent antinociceptive agents (ED50 ca. 0.020 microM/kg sc) and kappa ligands (Ki(kappa) ca. 0.20 nM) identified so far.

Analgesia↗

Inhibitors of topoisomerase II based on the benzodiimidazole and dipyrroloimidazobenzimidazole ring systems: controlling DT-diaphorase reductive inactivation with steric bulk.

Described herein are the synthesis, cytotoxic properties, and topoisomerase II inhibition assays of benzodiimidazole and dipyrroloimidazobenzimidazole structural variants of the pyrrolo[1, 2-a]benzimidazole or APBI ring system. These ring variants were designed to inhibit topoisomerase II, much as the APBIs are able to do. Since only the quinone form of the APBIs can intercalate DNA, two-electron reduction to the hydroquinone by DT-diaphorase is known to deactivate these compounds. Indeed, the APBIs possess a high inverse correlation with the cellular concentration of DT-diaphorase. Therefore one feature of the ABPI structural variants is the excessive bulk about the quinone ring, which was predicted to diminish DT-diaphorase substrate activity. Another feature is the presence of one or two alkylating centers, which would permit alkylation of DNA and/or topoisomerase II. Inhibition assays for topoisomerase II-mediated relaxation of supercoiled DNA indicate that the benzodiimidazole and dipyrroloimidazobenzimidazole quinone ring systems are catalytic inhibitors of topoisomerase II. Both quinone systems exhibit cytotoxicity perhaps due to the lack of inactivation by DT-diaphorase as well as topoisomerase II inhibition. One quinone displayed the novel feature of cytotoxicity selectively against melanoma cell lines. In conclusion, the benzodiimidazole and dipyrroloimidazobenzimidazole quinone ring systems will be subjected to future analogue development and structure-activity studies.

Animals↗

Regio- and Diastereoselective Catalytic Epoxidation of Acyclic Allylic Alcohols with Methyltrioxorhenium: A Mechanistic Comparison with Metal (Peroxy and Peroxo Complexes) and Nonmetal (Peracids and Dioxirane) Oxidants.

Geraniol and its 1-methyl derivative (regiochemical probes) and a set of methyl- and tert-butyl-substituted chiral allylic alcohols (stereochemical probes) have been used to elucidate the mechanism of the MTO-catalyzed epoxidation of allylic alcohols. The regiochemical probes are preferentially epoxidized at the unfunctionalized double bond by these MTO-based oxidants, which establishes that MTO/UHP and MTO/H(2)O(2)/pyridine mainly operate through hydrogen bonding. Metal-alcoholate binding does not apply, which is in contrast to the transition-metal oxidants VO(acac)(2)/t-BuOOH, Mo(CO)(6)/t-BuOOH, MoO(2)[PhCON(Ph)O](2)/t-BuOOH, MoO(O(2))[PhCON(Ph)O](2), and H(2)WO(4)/H(2)O(2). For the stereochemical probes, the diastereoselectivity data show a good correspondence between the MTO-catalyzed systems (MTO/UHP and MTO/H(2)O(2)/pyridine) and the perhydrate-type oxidant Ti-beta/H(2)O(2) and the peracid m-CPBA. Conformational control through 1,3-allylic strain results in a high threo diastereoselectivity, in which hydrogen bonding between the hydroxy functionality and the rhenium catalyst is the decisive electronic feature. All these selectivity data are consistent with the rhenium peroxo complex as the active oxidant.

Journal Article↗

Synthesis and incorporation into PNA of fluorinated olefinic PNA (F-OPA) monomers.

[structure: see text] A fluorinated OPA monomer containing the base thymine ((Z)-t-F-OPA) was synthesized in 12 steps, featuring a highly selective allylic over homoallylic Mitsunobu substitution for the introduction of the nucleobase. F-OPA modified PNA decamers were prepared by the MMTr/acyl protection strategy. The thermal stability of duplexes of PNA decamers containing (Z)-t-F-OPA units with antiparallel complementary DNA was measured. We found a strong dependence of stability from the sequential position of the (Z)-t-F-OPA units, ranging from DeltaT(m) of +2.4 to -8.1 degrees C/modification relative to unmodified PNA.

Alkenes↗

Intense immune suppression for systemic lupus--the role of hematopoietic stem cells.

The treatment of severe autoimmune diseases has been recently revitalized by the introduction of intense immune suppression with immune ablative intent followed by three different procedures. These are allogeneic hematopoietic stem cell transplantation (HSCT), autologous HSCT (using either marrow or peripheral blood), and intense immune suppression without stem cell support. Current trials suggest that high dose immune suppressive therapy with or without autologous hematopoietic stem cell support can induce remission of previously refractory disease. Follow-up is too brief to determine if intense immune suppression, and more specifically autologous HSCT, will ultimately cure SLE. It is conceivable that an allogeneic source of stem cells from a normal donor (e.g. HLA matched sibling) will be required to achieve a cure. It is also possible that autologous HSCT, even if not curative, may prolong the life of patients with otherwise high-risk features. In carefully selected patients, the potential benefits of this procedure may outweigh the risks.

Animals↗

Verbal pragmatics following unilateral stroke: emotional content and valence.

Verbal pragmatic aspects of discourse production were examined in 16 right brain-damaged (RBD), 16 left brain-damaged (LBD), and 16 normal control right-handed adults. The facilitation effect of emotional content, valence hypothesis, and relationship between pragmatics and emotion were evaluated. Participants produced monologues while recollecting emotional and nonemotional experiences. Transcribed monologues were rated for appropriateness on 6 pragmatic features: conciseness, lexical selection, quantity, relevancy, specificity, and topic maintenance. Overall, brain-damaged groups were rated as significantly less appropriate than normals. Consistent with the facilitation effect, emotional content enhanced pragmatic performance of LBD aphasic participants yet suppressed performance of RBD participants. Contrary to the valence hypothesis, RBD participants were more impaired for positive emotions and LBD participants for negative emotions. Pragmatic appropriateness was not strongly correlated with a measure of emotional intensity.

Adult↗

Making antibody fragments using phage display libraries.

To by-pass hybridoma technology and animal immunization, we are trying to build antibodies in bacteria by mimicking features of immune selection. Recently we used fd phage to display antibody fragments fused to a minor coat protein, allowing enrichment of phage with antigen. Using a random combinatorial library of the rearranged heavy (VH) and kappa (V kappa) light chains from mice immune to the hapten 2-phenyloxazol-5-one (phOx), we have now displayed diverse libraries of antibody fragments on the surface of fd phage. After a single pass over a hapten affinity column, fd phage with a range of phOx binding activities were detected, at least one with high affinity (dissociation constant, Kd = 10(-8) M). A second pass enriched for the strong binders at the expense of the weak. The binders were encoded by V genes similar to those found in anti-phOx hybridomas but in promiscuous combinations (where the same V gene is found with several different partners). By combining a promiscuous VH or V kappa gene with diverse repertoires of partners to create hierarchical libraries, we elicited many more pairings with strong binding activities. Phage display offers new ways of making antibodies from V-gene libraries, altering V-domain pairings and selecting for antibodies with good affinities.

Amino Acid Sequence↗

Chromogenic in situ hybridization-detected hotspot MYCN amplification associates with Ki-67 expression and inversely with nestin expression in neuroblastomas.

Since neuroblastomas are intratumorally heterogeneous, the analysis of genetic and biologic features of randomly selected tumor specimen spots may lead to erroneous conclusions. Our purpose was therefore to construct an easily assessable and strictly defined strategy to unify the detection of various molecular markers in paraffin-embedded neuroblastoma samples. We selected tumor specimen spots of highest proliferation activity, that is, hotspots, for the analysis of MYCN amplification status and proliferation-associated molecular markers, such as nestin, which role in neuroblastoma specimens was evaluated for the first time. Using a chromogenic in situ hybridization (CISH) technique, we showed that patients with a MYCN copy number higher than six in anti-Ki-67-detected hotspots have significantly worse overall survival prognosis than patients with low MYCN copy numbers (P = 0.0006). The chosen cutoff value of six was shown to dichotomize MYCN-amplified neuroblastomas at least as specifically as Southern blot hybridization, in which amplification was defined by a copy number of > or = 10. Interestingly, we also detected without difficulty MYCN-amplified neuroblastic cells in bone marrow samples using the CISH technique. The proliferation activity, assessed with an anti-Ki-67-based proliferation index, was significantly higher in MYCN-amplified than in nonamplified hotspots. The proliferation indices of the hotspots had also a significant correlation with the prognosis (International Classification) and histological type, whereas the proliferation accelerator Id2 did not associate with any of the mentioned parameters. The expression of nestin associated inversely with MYCN amplification (P = 0.018), which challenges a previously suggested role of nestin in neuroblastomas. In summary, hotspot focusing provides a means of analyzing proliferation-associated markers in neuroblastomas, and together with the CISH detection of the MYCN copy number enables an easy and reliable examination of MYCN status in neuroblastomas.

Antibodies, Antinuclear↗

Complex haplotypes, copy number polymorphisms and coding variation in two recently divergent mouse strains.

Inbred mouse strains provide the foundation for mouse genetics. By selecting for phenotypic features of interest, inbreeding drives genomic evolution and eliminates individual variation, while fixing certain sets of alleles that are responsible for the trait characteristics of the strain. Mouse strains 129Sv (129S5) and C57BL/6J, two of the most widely used inbred lines, diverged from common ancestors within the last century, yet very little is known about the genomic differences between them. By comparative genomic hybridization and sequence analysis of 129S5 short insert libraries, we identified substantial structural variation, a complex fine-scale haplotype pattern with a continuous distribution of diversity blocks, and extensive nucleotide variation, including nonsynonymous coding SNPs and stop codons. Collectively, these genomic changes denote the level and direction of allele fixation that has occurred during inbreeding and provide a basis for defining what makes these mouse strains unique.

Animals↗